Name: duodenal-jejunal bypass surgery in diet-induced obese diabetic mice
Uploaded: 2024-10-18T12:40:00
Description: The prevalence of obesity and type 2 diabetes is a serious global health concern. Obesity is a major pathogenic factor in type 2 diabetes, cardiovascular ...

This study was supported by a grant from the Science and Technology Planning Project of Guangdong Province of China (No. 202002020069).

All protocol steps described below follow the guidelines of the Animal Care and Use Committee of the General Hospital of the Southern Theater Command under the approval number 2020112501.
1. General preoperative preparation
NOTE: Thirty 6-week-old male C57BL/6 mice were purchased. Mice were housed in a Specific Pathogen-Free (SPF) laboratory under a 12 h light/dark cycle; temperature, 22 &#177; 2 &#176;C; and humidity, 55-65%. Mice were given free acc...

<ol>
	<li>Lascar, N., Brown, J., Pattison, H., Barnett, A. H., Bailey, C. J., Bellary, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Type+2+diabetes+in+adolescents+and+young+adults.">Type 2 diabetes in adolescents and young adults.</a> Lancet Diabetes Endocrinol. 6 (1), 69-80 (2018).</li><li>Song, Y., Zhao, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bibliometric+analysis+of+metabolic+surgery+for+type+2+diabetes:+current+status+and+future+prospects.">Bibliometric analysis of metabolic surgery for type 2 diabetes: current status and future prospects.</a> Updates Surg. 74 (2), 697-707 (2022).</li><li>Cummings, D. 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W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanisms+of+weight+loss+after+obesity+surgery.">Mechanisms of weight loss after obesity surgery.</a> Endocr Rev. 43 (1), 19-34 (2022).</li><li>Rubino, F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+mechanism+of+diabetes+control+after+gastrointestinal+bypass+surgery+reveals+a+role+of+the+proximal+small+intestine+in+the+pathophysiology+of+type+2+diabetes.">The mechanism of diabetes control after gastrointestinal bypass surgery reveals a role of the proximal small intestine in the pathophysiology of type 2 diabetes.</a> Ann Surg. 244 (5), 741-749 (2006).</li><li>Rubino, F., Marescaux, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+duodenal-jejunal+exclusion+in+a+non-obese+animal+model+of+type+2+diabetes:+a+new+perspective+for+an+old+disease.">Effect of duodenal-jejunal exclusion in a non-obese animal model of type 2 diabetes: a new perspective for an old disease.</a> Ann Surg. 239 (1), 1-11 (2004).</li><li>Akalestou, E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Establishing+a+successful+rat+model+of+duodenal-+jejunal+bypass:+A+detailed+guide.">Establishing a successful rat model of duodenal- jejunal bypass: A detailed guide.</a> Lab Anim. 53 (4), 362-371 (2019).</li><li>Yu, H. H., Hsieh, M. C., Wu, S. Y., Sy, E. D., Shan, Y. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+duodenal-jejunal+bypass+surgery+in+ameliorating+nonalcoholic+steatohepatitis+in+diet-induced+obese+rats.">Effects of duodenal-jejunal bypass surgery in ameliorating nonalcoholic steatohepatitis in diet-induced obese rats.</a> Diabetes Metab Syndr Obes. 12, 149-159 (2019).</li><li>Garciacaballero, M., Navarrete, S., Favretti, F., Celik, A., Del Castillo, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diabetes+surgery+in+type+2+BMI+24-29+vs+IMC+30-34+diabetic+patients:+is+there+differences+among+restrictive,+malabsorptive+and+gastric+bypass+procedures.">Diabetes surgery in type 2 BMI 24-29 vs IMC 30-34 diabetic patients: is there differences among restrictive, malabsorptive and gastric bypass procedures.</a> Nutr Hosp. 28 (2), 23-30 (2013).</li><li>Vidal, J., Jim&#233;nez, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diabetes+remission+following+metabolic+surgery:+is+GLP-1+the+culprit.">Diabetes remission following metabolic surgery: is GLP-1 the culprit.</a> Curr Atheroscler Rep. 15 (10), 357 (2013).</li><li>Speck, M., Cho, Y. M., Asadi, A., Rubino, F., Kieffer, T. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Duodenal-jejunal+bypass+protects+GK+rats+from+{beta}-cell+loss+and+aggravation+of+hyperglycemia+and+increases+enteroendocrine+cells+coexpressing+GIP+and+GLP-1.">Duodenal-jejunal bypass protects GK rats from {beta}-cell loss and aggravation of hyperglycemia and increases enteroendocrine cells coexpressing GIP and GLP-1.</a> Am J Physiol Endocrinol Metab. 300 (5), E923-E932 (2011).</li><li>Ashrafian, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Metabolic+surgery:+an+evolution+through+bariatric+animal+models.">Metabolic surgery: an evolution through bariatric animal models.</a> Obes Rev. 11 (12), 907-920 (2010).</li><li>Rao, R. S., Rao, V., Kini, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+models+in+bariatric+surgery--a+review+of+the+surgical+techniques+and+postsurgical+physiology.">Animal models in bariatric surgery--a review of the surgical techniques and postsurgical physiology.</a> Obes Surg. 20 (9), 1293-1305 (2010).</li><li>Im, Y. R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+systematic+review+of+animal+models+of+NAFLD+finds+high-fat,+high-fructose+diets+most+closely+resemble+human+NAFLD.">A systematic review of animal models of NAFLD finds high-fat, high-fructose diets most closely resemble human NAFLD.</a> Hepatology. 74 (4), 1884-1901 (2021).</li><li>Kleinert, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+models+of+obesity+and+diabetes+mellitus.">Animal models of obesity and diabetes mellitus.</a> Nat Rev Endocrinol. 14 (3), 140-162 (2018).</li><li>Furman, B. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Streptozotocin-induced+diabetic+models+in+mice+and+rats.">Streptozotocin-induced diabetic models in mice and rats.</a> Curr Protoc. 1 (4), e78 (2021).</li><li>Buchwald, H., Varco, R. L., Moore, R. B., Schwartz, M. Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intestinal+bypass+procedures.+Partial+ileal+bypass+for+hyperlipidemia+and+jejunoileal+bypass+for+obesity.">Intestinal bypass procedures. Partial ileal bypass for hyperlipidemia and jejunoileal bypass for obesity.</a> Curr Probl Surg. , 1-51 (1975).</li><li>DeMaria, E. 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E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gastric+bypass+for+morbid+obesity.">Gastric bypass for morbid obesity.</a> Surg Annu. 11, 99-126 (1979).</li><li>Cheng, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sleeve+gastrectomy+with+bypass+of+proximal+small+intestine+provides+better+diabetes+control+than+sleeve+gastrectomy+alone+under+postoperative+high-fat+diet.">Sleeve gastrectomy with bypass of proximal small intestine provides better diabetes control than sleeve gastrectomy alone under postoperative high-fat diet.</a> Obes Surg. 29 (1), 84-92 (2019).</li><li>Rubino, F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+role+of+the+small+bowel+in+the+regulation+of+circulating+ghrelin+levels+and+food+intake+in+the+obese+Zucker+rat.">The role of the small bowel in the regulation of circulating ghrelin levels and food intake in the obese Zucker rat.</a> Endocrinology. 146 (4), 1745-1751 (2005).</li><li>Zachariah, P. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Compared+to+sleeve+gastrectomy,+duodenal-jejunal+bypass+with+sleeve+gastrectomy+gives+better+glycemic+control+in+T2DM+patients,+with+a+lower+&#946;-cell+response+and+similar+appetite+sensations:+mixed-meal+study.">Compared to sleeve gastrectomy, duodenal-jejunal bypass with sleeve gastrectomy gives better glycemic control in T2DM patients, with a lower &#946;-cell response and similar appetite sensations: mixed-meal study.</a> Obes Surg. 26 (12), 2862-2872 (2016).</li><li>Jiang, B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Role+of+proximal+intestinal+glucose+sensing+and+metabolism+in+the+blood+glucose+control+in+type+2+diabetic+rats+after+duodenal+jejunal+bypass+surgery.">Role of proximal intestinal glucose sensing and metabolism in the blood glucose control in type 2 diabetic rats after duodenal jejunal bypass surgery.</a> Obes Surg. 32 (4), 1119-1129 (2022).</li><li>Han, H. F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Duodenal-jejunal+bypass+increases+intraduodenal+bile+acids+and+upregulates+duodenal+SIRT1+expression+in+high-fat+diet+and+streptozotocin-induced+diabetic+rats.">Duodenal-jejunal bypass increases intraduodenal bile acids and upregulates duodenal SIRT1 expression in high-fat diet and streptozotocin-induced diabetic rats.</a> World J Gastroenterol. 28 (31), 4338-4350 (2022).</li><li>Yan, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reduction+of+intestinal+electrogenic+glucose+absorption+after+duodenojejunal+bypass+in+a+mouse+model.">Reduction of intestinal electrogenic glucose absorption after duodenojejunal bypass in a mouse model.</a> Obes Surg. 23 (9), 1361-1369 (2013).</li><li>Arble, D. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Metabolic+comparison+of+one-anastomosis+gastric+bypass,+single-anastomosis+duodenal-switch,+Roux-en-Y+gastric+bypass,+and+vertical+sleeve+gastrectomy+in+rat.">Metabolic comparison of one-anastomosis gastric bypass, single-anastomosis duodenal-switch, Roux-en-Y gastric bypass, and vertical sleeve gastrectomy in rat.</a> Surg Obes Relat Dis. 14 (12), 1857-1867 (2018).</li><li>Liang, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Duodenal-jejunal+bypass+surgery+reverses+diabetic+phenotype+and+reduces+obesity+in+db/db+Mice.">Duodenal-jejunal bypass surgery reverses diabetic phenotype and reduces obesity in db/db Mice.</a> Curr Chem Genom Transl Med. 11, 41-49 (2017).</li><li>Barataud, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Absence+of+role+of+dietary+protein+sensing+in+the+metabolic+benefits+of+duodenal-jejunal+bypass+in+the+mouse.">Absence of role of dietary protein sensing in the metabolic benefits of duodenal-jejunal bypass in the mouse.</a> Sci Rep. 7, 44856 (2017).</li><li>. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Close-up+on+lab+animal+microsurgery.">Close-up on lab animal microsurgery.</a> Lab Anim (NY). 35 (1), 43 (2006).</li><li>Couceiro, J., Castro, R., Tien, H., Ozyurekoglu, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Step+by+step:+microsurgical+training+method+combining+two+nonliving+animal+models.">Step by step: microsurgical training method combining two nonliving animal models.</a> J Vis Exp. (99), e52625 (2015).</li><li>Nasser, K. M., Wahba, H. A., Kamal, E., El-Makhzangy, A. M., Bahaa, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+model+for+training+and+improvement+of+the+surgical+skills+in+endolaryngeal+microsurgery.">Animal model for training and improvement of the surgical skills in endolaryngeal microsurgery.</a> J Voice. 26 (3), 351-357 (2012).</li></ol>

In 1953, Varco et al.18 performed the first jejuno-ileal bypass as the beginning of bariatric surgery. Since then, numerous bariatric surgeries have been performed by surgeons. These surgeries have resulted in weight loss and improved metabolic complications4,19,20. Furthermore, in 1967, Mason and Ito21 performed the first Roux-en-Y gastric bypass (RYGB), which can effectively redu...

Obesity and type 2 diabetes are the most common chronic diseases in the world, and their prevalence is increasing among young individuals1. Bariatric surgery is the most effective treatment for obesity and diabetes, facilitating long-term blood glucose stabilitywhile also improving the complications associated with obesity2,3. There are several types of bariatric surgery, classified by whether they reduce gastric volume or intestinal absorption; these include restrictive, malabsorptive, and combination4,5.
Duodenal-jejunal bypass (DJB) was first developed by Rubino and Marescaux, who demonstrated that type 2 diabetes could be alleviated by connecting the duodenum and jejunum rather than by reducing the gastric volume6,7. DJB preserves the entire stomach and bypasses the entire duodenum and proximal jejunum. The bowel is divided into biliopancreatic, digestive, and common limbs6,8. DJB shares some similarities with bariatric surgeries, including Roux-en-Y gastric bypass (RYGB), mini-gastric bypass, biliopancreatic bypass, duodenal diversion, and DJB plus sleeve gastrectomy9. Compared to RYGB, DJB does not require a gastrointestinal anastomosis, which reduces the operative time and improves the success rate of the procedure. DJB is similar to RYGB in improving glucose metabolism but does not affect body weight10. After DJB surgery, rapid food delivery to the distal small intestine stimulates the secretion of glucagon-like peptide-1 (GLP-1), resulting in improved glucose metabolism11,12.
The use of animal models is essential for understanding the metabolic, cellular, and molecular pathways. Animal models of bariatric surgery have contributed to our understanding of potential mechanisms underlying obesity and diabetes13,14. However, due to the physiological differences between species, it is impossible to perfectly replicate human diseases in animal models15. Among various animal models available for research purposes, the diet-induced obesity (DIO) mouse model most closely resembles human obesity and metabolic syndrome16. Mice were selected for DJB surgery to determine the feasibility of the surgery and to provide techniques for further research. This manuscript provides a comprehensive summary of both technical aspects and experimental details of DJB surgery.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Abdominal retractor</td>
			<td>F.S.T</td>
			<td>17000-03</td>
			<td>Colibri Retractor -3cm,retractor range 1.5cm/3cm long</td>
		</tr>
		<tr>
			<td>1% sodium pentobarbital solution</td>
			<td>Guangzhou Chemical Reagent Factory</td>
			<td>/</td>
			<td>&#160;Dissolved 500mg of pentobarbital sodium powder in 50ml of normal saline to obtain 1% pentobarbital sodium solution.</td>
		</tr>
		<tr>
			<td>Benzylpenicillin sodium for Injection</td>
			<td>North China Pharmaceutical Company Ltd.</td>
			<td>F2062121</td>
			<td>Penicillin</td>
		</tr>
		<tr>
			<td>Buprenorphine</td>
			<td>Guangzhou Chemical Reagent Factory</td>
			<td>/</td>
			<td>Analgesia</td>
		</tr>
		<tr>
			<td>Buprenorphine</td>
			<td>Guangzhou Otsuka Pharmaceutical Co., Ltd</td>
			<td>/</td>
			<td>Analgesic</td>
		</tr>
		<tr>
			<td>Citric acid-sodium citrate buffer</td>
			<td>LEAGENE</td>
			<td>R00522</td>
			<td>Buffer solution</td>
		</tr>
		<tr>
			<td>Cotton buds</td>
			<td>HaoZheng Medical</td>
			<td>60220610</td>
			<td>Cotton swabs</td>
		</tr>
		<tr>
			<td>Depilatory paste</td>
			<td>Veet</td>
			<td>AAPR-S222</td>
			<td>Hair removal cream</td>
		</tr>
		<tr>
			<td>Ear tag</td>
			<td>ZEYA</td>
			<td>SUS304</td>
			<td>Ear-mark</td>
		</tr>
		<tr>
			<td>Electric blanket</td>
			<td>ZOSEN</td>
			<td>ZS-CWDRT</td>
			<td>Heat pad</td>
		</tr>
		<tr>
			<td>Electronic scale</td>
			<td>WETTLER TOLEDO</td>
			<td>20060902-6</td>
			<td>Measure the weight</td>
		</tr>
		<tr>
			<td>Enteral nutritional powder</td>
			<td>Abbott Laboratories</td>
			<td>/</td>
			<td>Nutrition powder</td>
		</tr>
		<tr>
			<td>Eye ointment</td>
			<td>Guangzhou Otsuka Pharmaceutical Co., Ltd</td>
			<td>/</td>
			<td>Protect the eyes</td>
		</tr>
		<tr>
			<td>Glucometer</td>
			<td>Roche</td>
			<td>6993788001</td>
			<td>Assess blood glucose</td>
		</tr>
		<tr>
			<td>Graphpad Prism version 9.4.1</td>
			<td>GraphPad Software</td>
			<td>version 9.4.1</td>
			<td>Software for statistical analysis</td>
		</tr>
		<tr>
			<td>High-fat diet (High Fat [60FDC] Purified Rodent Diet)</td>
			<td>Dyets</td>
			<td>112252</td>
			<td>60kcal% High Fat Diet</td>
		</tr>
		<tr>
			<td>Micro Forceps</td>
			<td>Jinzhong Medical</td>
			<td>18-1140</td>
			<td>Micro forceps</td>
		</tr>
		<tr>
			<td>Micro needle holder</td>
			<td>Jinzhong Medical</td>
			<td>EMT-160-Z</td>
			<td>Needle holder</td>
		</tr>
		<tr>
			<td>Micro Scissors</td>
			<td>Jinzhong Medical</td>
			<td>YBC010</td>
			<td>Micro scissors</td>
		</tr>
		<tr>
			<td>Microscope camera</td>
			<td>LAPSUN</td>
			<td>E 2000</td>
			<td>Video</td>
		</tr>
		<tr>
			<td>Ophthalmic scissors</td>
			<td>Jinzhong Medical</td>
			<td>Y00030</td>
			<td>Surgical scissors</td>
		</tr>
		<tr>
			<td>Pentobarbital</td>
			<td>Guangzhou Chemical Reagent Factory</td>
			<td>/</td>
			<td>Narcosis</td>
		</tr>
		<tr>
			<td>Sodium chloride Injection</td>
			<td>Guangzhou Otsuka Pharmaceutical Co., Ltd</td>
			<td>B21L0301</td>
			<td>NaCl 0.9%</td>
		</tr>
		<tr>
			<td>Stereo microscope</td>
			<td>ZEISS</td>
			<td>Stemi 305</td>
			<td>Binocular stereomicroscope</td>
		</tr>
		<tr>
			<td>Streptozotocin</td>
			<td>Sigma</td>
			<td>S110910-1g</td>
			<td>STZ</td>
		</tr>
		<tr>
			<td>Suture line</td>
			<td>LINGQIAO SUTURE</td>
			<td>ZS-LQPMRZ5/0</td>
			<td>Prolene 6/0,Prolene 10/0</td>
		</tr>
		<tr>
			<td>Tissue forceps</td>
			<td>Jinzhong Medical</td>
			<td>H1701</td>
			<td>Surgical forceps</td>
		</tr>
	</tbody>
</table>

duodenal-jejunal bypass surgery in diet-induced obese diabetic mice

The prevalence of obesity and type 2 diabetes is a serious global health concern. Obesity is a major pathogenic factor in type 2 diabetes, cardiovascular disease, and some cancers. Bariatric surgery offers a long-term and effective treatment option for both obesity and diabetes. Sleeve gastrectomy (SG) and Roux-en-Y gastric bypass (RYGB) are widely recognized as the most popular bariatric surgeries. Additionally, several exploratory bariatric surgeries have demonstrated promising therapeutic effects. Duodenal-jejunal bypass (DJB), specifically tailored for diabetics with low body mass index, has shown beneficial metabolic outcomes. However, its weight-independent metabolic benefits are not fully understood due to limited animal models. In this article, we describe the optimized care protocols and surgical techniques for performing DJB surgery in diet-induced obese (DIO) diabetic mice. Using a mouse model contributes to a better understanding of the nature of changes induced by DJB surgery while facilitating related clinical practice.

General conditions 
The mean operative time for the DJB procedure was 84.5 &#177; 2.6 min. Fifteen mice underwent DJB surgery, nine mice survived. As shown in Table 1, the majority of deaths occurred during surgery or in the following 7 days. The causes of postoperative death were bleeding (n=2) on postoperative day 1, anastomotic leakage (n=1) on postoperative day 4, anastomotic obstruction (n=2) on postoperative day 7, and unknown cause (n=1) three weeks after surgery.
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Duodenal-jejunal bypass (DJB) surgery can improve glucose metabolism and reduce insulin resistance. Here, we present a protocol to establish a stable and reliable mouse model of DJB.

Duodenal-Jejunal Bypass Surgery in Diet-Induced Obese Diabetic Mice

The prevalence of obesity and type 2 diabetes is a serious global health concern. Obesity is a major pathogenic factor in type 2 diabetes, cardiovascular ...

The increasing instance of obesity and Type 2 Diabetes has become a serious health problem. Duodenal-jejunal bypass surgery, DJB, can improve glucose metabolism and ameliorate insulin resistance. And its metabolic benefits independent of weight loss are not fully understood.Diet-induced obese diabetical mice were selected for DJB surgery to determine the feasibility and vertical structure of the surgery and to provide technicals for subsequent research. Mice were injected intraperitoneally with 1%pentobarbital sodium solution and buprenorphine. After anesthetizing the mice, the toe pinch reflex was checked, placed on the board.And then the depilatory cream was applied to the abdomen of the mice. Disinfect the skin three times with iodophor and ethyl alcohol. Cover the animal with a sterile drape and make a two centimeter incision from the xiphoid to the abdomen.Use an abdominal retractor to expose the abdominal cavity. The jejunal five centimeter distal to the ligament of Treitz was double ligated with six zero silk suture. Cut the jejunum in the middle of the two ligations.Pull the proximal jejunal incision five centimeter along the bowel to the jejunum to create the jejunojejunostomy. The anastomotic works of the two bowels were aligned horizontally and they used a 10 zero silk suture for natural enteromosis. Fixed force ends of the bowels and cut the incision in this.Suture the second layer of the posterior intestinal wall with a four simple continuous suture, fixed both ends of the bowel and suture the anterior wall. Sutures the fourth layer of the anterior wall with a simple continuous suture and suture the thick layer of the plasma muscle with a horizontal various. Pull the distal jejunal incision to the duodenal one centimeter below the panerus and make the duodenal jejunal stormy in the same method.Double-neck the duodenal with a 606 suture, two millimeter from the distal end of the duodenal jejunal and enteromosis. Cut in the middle and suture the dump with a 10 zero silk suture. Rearrange and return the intestines to the abdominal cavity.Raise the abdominal cavity with 13 centigrade saline. Suture the muscle and the skin separately with a six zero silk. Disinfect skin with iodophor.After the operation, inject saline and penicillin to prevent dehydration and infection. Inject buprenorphine for postoperative analgesia. After surgery, placed mouse on the electric blanket to prevent hyperthermia until it was awake.On the postoperative day, food and water was restricted. On the first postoperative day, give the mice sugar water drink, observe the health of the mice daily. We can see that the survival rate is gradually increasing, indicating that we need intensive surgical training to master the DJB technique.Mouse had similar body weight and daily food intakes before and eight weeks after surgery. DJB mice did not show significant weight loss compared to SHAM control mice at eight weeks after surgery. Benefit intake was reduced because of dietary transition during the seven day period.Forming DJB and SHAM control surgery and returned to the preoperative level three weeks later. By measuring the blood glucose, we can see that the blood glucose of the mice decreased significantly after the DJB surgery and the glucose tolerance was also significantly improved. I believe that through the previous introduction you have a physical understanding of DJB surgery and have a full understanding of its improvement of blood glucose.We can master this technology within 18 minutes. The most important thing is to remember all the procedures and be proficient in this technology. For example, the suturing process should be strict.In our procedure, we believe that preoperative preparation and postoperative care play an important role in the entire surgical process. It could have answered key questions about obesity and metabolism and provide a pathway for procedures in related fields. We hope this technology will help you.

duodenal-jejunal-bypass-surgery-in-diet-induced-obese-diabetic-mice

Research

JoVE Journal

Medicine

Roux-en-Y Gastric Bypass Operation in Rats

Currently, the most effective therapy for the treatment of morbid obesity to induce significant and maintained body weight loss with a proven mortality benefit is bariatric surgery1,2. Consequently, there has been a steady rise in the number of bariatric operations done worldwide in recent years with the Roux-en-Y gastric bypass (gastric bypass) being the most commonly performed operation3. Against this background, it is important to understand the physiological mechanisms by which gastric bypass induces and maintains body weight loss. These mechanisms are yet not fully understood, but may include reduced hunger and increased satiation4,5, increased energy expenditure6,7, altered preference for food high in fat and sugar8,9, altered salt and water handling of the kidney10 as well as alterations in gut microbiota11. Such changes seen after gastric bypass may at least partly stem from how the surgery alters the hormonal milieu because gastric bypass increases the postprandial release of peptide-YY (PYY) and glucagon-like-peptide-1 (GLP-1), hormones that are released by the gut in the presence of nutrients and that reduce eating12.
During the last two decades numerous studies using rats have been carried out to further investigate physiological changes after gastric bypass. The gastric bypass rat model has proven to be a valuable experimental tool not least as it closely mimics the time profile and magnitude of human weight loss, but also allows researchers to control and manipulate critical anatomic and physiologic factors including the use of appropriate controls. Consequently, there is a wide array of rat gastric bypass models available in the literature reviewed elsewhere in more detail 13-15. The description of the exact surgical technique of these models varies widely and differs e.g. in terms of pouch size, limb lengths, and the preservation of the vagal nerve. If reported, mortality rates seem to range from 0 to 35%15. Furthermore, surgery has been carried out almost exclusively in male rats of different strains and ages. Pre- and postoperative diets also varied significantly.
Technical and experimental variations in published gastric bypass rat models complicate the comparison and identification of potential physiological mechanisms involved in gastric bypass. There is no clear evidence that any of these models is superior, but there is an emerging need for standardization of the procedure to achieve consistent and comparable data. This article therefore aims to summarize and discuss technical and experimental details of our previously validated and published gastric bypass rat model.

Numerous studies using gastric bypass rat models have been recently conducted to uncover the underlying physiological mechanisms of Roux-en-Y gastric bypass operations. This article aims to demonstrate and discuss the technical and experimental details of our published gastric bypass rat model to understand advantages and limitations of this experimental tool.

Currently, the most effective therapy for the treatment of morbid obesity to induce significant and maintained body weight loss with a proven mortality ...

Assessment of Gastric Emptying in Non-obese Diabetic Mice Using a [13C]-octanoic Acid Breath Test

Gastric emptying studies in mice have been limited by the inability to follow gastric emptying changes in the same animal since the most commonly used techniques require killing of the animals and postmortem recovery of the meal1,2. This approach prevents longitudinal studies to determine changes in gastric emptying with age and progression of disease. The commonly used [13C]-octanoic acid breath test for humans3 has been modified for use in mice4-6 and rats7 and we previously showed that this test is reliable and responsive to changes in gastric emptying in response to drugs and during diabetic disease progression8. In this video presentation the principle and practical implementation of this modified test is explained. As in the previous study, NOD LtJ mice are used, a model of type 1 diabetes9. A proportion of these mice develop the symptoms of gastroparesis, a complication of diabetes characterized by delayed gastric emptying without mechanical obstruction of the stomach10.
This paper demonstrates how to train the mice for testing, how to prepare the test meal and obtain 4 hr gastric emptying data and how to analyze the obtained data. The carbon isotope analyzer used in the present study is suitable for the automatic sampling of the air samples from up to 12 mice at the same time. This technique allows the longitudinal follow-up of gastric emptying from larger groups of mice with diabetes or other long-standing diseases.

Assessment of Gastric Emptying in Non-obese Diabetic Mice Using a [13C]-octanoic Acid Breath Test

Determination of gastric emptying with a non-invasive [13C]-octanoic acid breath test for tracking gastroparesis in female NOD LtJ mice.

Gastric emptying studies in mice have been limited by the inability to follow gastric emptying changes in the same animal since the most commonly used ...

Vein Interposition Model: A Suitable Model to Study Bypass Graft Patency

Bypass grafting is an established treatment method for coronary artery disease. Graft patency continues to be the Achilles heel of saphenous vein grafts. Research models for bypass graft failure are essential for a better understanding of pathobiological and pathophysiological processes during graft patency loss. Large animal models, such as pigs or sheep, resemble human anatomical structures but require special facilities and equipment. This video describes a rat vein interposition model to investigate vein graft patency loss. Rats are inexpensive and easy to handle. Compared to mouse models, the convenient size of rats permits better operability and enables a sufficient amount of material to be obtained for further diverse analysis. In brief, the inferior epigastric vein of a donor rat is harvested and used to replace a segment of the femoral artery. Anastomosis is conducted via single stitches and sealed with fibrin glue. Graft patency can be monitored non-invasively using duplex sonography. Myointimal hyperplasia, which is the main cause for graft patency loss, develops progressively over time and can be calculated from histological cross sections.

This video demonstrates a model to study the development of myointimal hyperplasia after venous interposition surgery in rats.

Bypass grafting is an established treatment method for coronary artery disease. Graft patency continues to be the Achilles heel of saphenous vein grafts. ...

Techniques of Sleeve Gastrectomy and Modified Roux-en-Y Gastric Bypass in Mice

Obesity is a major public health issue, with a prevalence of 4 to 28% for men and 6.2 to 36.5% for women in Europe (from 2003 to 2008). Morbid obesity is frequently associated with metabolic complications, such as type 2 diabetes, hypertension, and dyslipidemia, reducing life expectancy and quality. In the absence of any effective noninvasive treatments, bariatric surgery is a valuable therapeutic option for patients with morbid obesity (body mass index (BMI) &#62;40 kg/m2), leading to long-term, sustained weight loss and improvements in metabolic complications. However, the underlying cellular and molecular mechanisms sustaining the beneficial effects of bariatric surgery are not yet fully understood. Due to the numerous genetically-modified strains available, the mouse model is the most convenient animal model to explore the molecular mechanisms behind the pleiotropic beneficial effects of bariatric surgeries. Here, we detailed the optimized healthcare methods and surgical protocols in mice for the two most widely-used bariatric surgeries: the sleeve gastrectomy and the modified Roux-en-Y gastric bypass. Deciphering the molecular mechanisms underlying the therapeutic effects of bariatric surgeries offers the promise of identifying new therapeutics targets.

Bariatric surgery is the most efficient way to reduce body weight and the deadly metabolic complications (diabetes, obesity, and dyslipidemia) frequently associated with morbid obesity. Mouse models of bariatric surgery represent a unique asset for deciphering molecular mechanisms behind the beneficial effects of these surgeries on diabetes, hypertension, and dyslipidemia.

Obesity is a major public health issue, with a prevalence of 4 to 28% for men and 6.2 to 36.5% for women in Europe (from 2003 to 2008). Morbid obesity is ...

Cardiopulmonary Bypass in a Mouse Model: A Novel Approach

As prolonged cardiopulmonary bypass becomes more essential during cardiac interventions, an increasing clinical demand arises for procedure optimization and for minimizing organ damage resulting from prolonged extracorporal circulation. The goal of this paper was to demonstrate a fully functional and clinically relevant model of cardiopulmonary bypass in a mouse. We report on the device design, perfusion circuit optimization, and microsurgical techniques. This model is an acute model, which is not compatible with survival due to the need for multiple blood drawings. Because of the range of tools available for mice (e.g., markers, knockouts, etc.), this model will facilitate investigation into the molecular mechanisms of organ damage and the effect of cardiopulmonary bypass in relation to other comorbidities.

This paper describes how to perform cardiopulmonary bypass in mice. This novel model will facilitate the investigation of the molecular mechanisms involved in organ damage.

As prolonged cardiopulmonary bypass becomes more essential during cardiac interventions, an increasing clinical demand arises for procedure optimization ...

An Experimental Model of Diet-Induced Metabolic Syndrome in Rabbit: Methodological Considerations, Development, and Assessment

In recent years, obesity and metabolic syndrome (MetS) have become a growing problem for public health and clinical practice, given their increased prevalence due to the rise of sedentary lifestyles and unhealthy eating habits. Thanks to animal models, basic research can investigate the mechanisms underlying pathological processes such as MetS. Here, we describe the methods used to develop an experimental rabbit model of diet-induced MetS and its assessment. After a period of acclimation, animals are fed a high-fat (10% hydrogenated coconut oil and 5% lard), high-sucrose (15% sucrose dissolved in water) diet for 28 weeks. During this period, several experimental procedures were performed to evaluate the different components of MetS: morphological and blood pressure measurements, glucose tolerance determination, and the analysis of several plasma markers. At the end of the experimental period, animals developed central obesity, mild hypertension, pre-diabetes, and dyslipidemia with low HDL, high LDL, and an increase of triglyceride (TG) levels, thus reproducing the main components of human MetS. This chronic model allows new perspectives for understanding the underlying mechanisms in the progression of the disease, the detection of preclinical and clinical markers that allow the identification of patients at risk, or even the testing of new therapeutic approaches for the treatment of this complex pathology.

We describe methods to develop an experimental model of diet-induced metabolic syndrome (MetS) in rabbits using a high-fat, high-sucrose diet. Animals developed central obesity, mild hypertension, pre-diabetes, and dyslipidemia, thus reproducing the main components of human MetS. This chronic model will allow acquisition of knowledge underlying mechanisms of disease progression.

In recent years, obesity and metabolic syndrome (MetS) have become a growing problem for public health and clinical practice, given their increased ...

Surgical Swine Model of Chronic Cardiac Ischemia Treated by Off-Pump Coronary Artery Bypass Graft Surgery

Chronic cardiac ischemia that impairs cardiac function, but does not result in infarct, is termed hibernating myocardium (HM). A large clinical subset of coronary artery disease (CAD) patients have HM, which in addition to causing impaired function, puts them at higher risk for arrhythmia and future cardiac events. The standard treatment for this condition is revascularization, but this has been shown to be an imperfect therapy. The majority of pre-clinical cardiac research focuses on infarct models of cardiac ischemia, leaving this subset of chronic ischemia patients largely underserved. To address this gap in research, we have developed a well-characterized and highly reproducible model of hibernating myocardium in swine, as swine are ideal translational models for human heart disease. In addition to creating this unique disease model, we have optimized a clinically relevant treatment model of coronary artery bypass surgery in swine. This allows us to accurately study the effects of bypass surgery on heart disease, as well as investigate additional or alternate therapies. This model surgically induces single vessel stenosis by implanting a constrictor on the left anterior descending (LAD) artery in a young pig. As the pig grows, the constrictor creates a gradual stenosis, resulting in chronic ischemia with impaired regional function, but preserving tissue viability. Following the establishment of the hibernating myocardium phenotype, we perform off-pump coronary artery bypass graft surgery to revascularize the ischemic region, mimicking the gold-standard treatment for patients in the clinic.

This protocol presents a surgical large animal model of chronic, single vessel ischemia that results in regional abnormalities but does not create infarct, known as hibernating myocardium. Following establishment of chronic ischemia, animals are treated with off-pump LIMA-LAD coronary artery bypass graft surgery to revascularize the ischemic tissue.

Chronic cardiac ischemia that impairs cardiac function, but does not result in infarct, is termed hibernating myocardium (HM). A large clinical subset of ...

Protocol to Create Chronic Wounds in Diabetic Mice

Chronic wounds develop as a result of defective regulation in one or more complex cellular and molecular processes involved in proper healing. They impact ~6.5M people and cost ~$40B/year in the US alone. Although a significant effort has been invested in understanding how chronic wounds develop in humans, fundamental questions remain unanswered. Recently, we developed a novel mouse model for diabetic chronic wounds that have many characteristics of human chronic wounds. Using db/db-/- mice, we can generate chronic wounds by inducing high levels of oxidative stress (OS) in the wound tissue immediately after wounding, using a one-time treatment with inhibitors specific to the antioxidant enzymes catalase and glutathione peroxidase. These wounds have high levels of OS, develop biofilm naturally, become fully chronic within 20 days after treatment and can remain open more for more than 60 days. This novel model has many features of diabetic chronic wounds in humans and therefore can contribute significantly to advancing fundamental understanding of how wounds become chronic. This is a major breakthrough because chronic wounds in humans cause significant pain and distress to patients and result in amputation if unresolved. Moreover, these wounds are very expensive and time-consuming to treat, and lead to significant loss of personal income to patients. Advancements in this field of study through the use of our chronic wound model can significantly improve health care for millions who suffer under this debilitating condition. In this protocol, we describe in great detail the procedure to cause acute wounds to become chronic, which has not been done before.

Chronic wounds are developed from acute wounds on a diabetic mouse model by inducing high levels of oxidative stress after a full-thickness cutaneous wound. The wound is treated with inhibitors for catalase and glutathione peroxidase, resulting in impaired healing and biofilm development by bacteria present in the skin microbiome.

Chronic wounds develop as a result of defective regulation in one or more complex cellular and molecular processes involved in proper healing. They impact ...

One-anastomosis Gastric Bypass (OAGB) in Rats

The goal of this protocol is to set up a preclinical model of bariatric surgery and, more specifically, OAGB in obese rats. Based on this preclinical model, longitudinal studies can be carried out to provide an improved understanding of the mechanisms underlying the outcomes seen after bariatric surgery in humans. For this purpose, rats are operated on through a laparotomy under general anesthesia with isoflurane. First, the surgeon creates a long and tubular gastric pouch: after greater curve and hiatal dissection, the nonglandular stomach is stapled and removed. Then, the remaining stomach is also stapled in order to create a gastric tube and exclude the antrum of the stomach. After that, the surgeon performs a single end-to-side gastrojejunostomy 35 cm from the duodenojejunal angle. This limb length has been chosen in order to reproduce the same ratio between the biliopancreatic limb (BPL) and common limb (CL) length as in human bariatric surgery. The operation ends by aponeurotic and cutaneous closure. The early postoperative management consists of subcutaneous hydration, an intramuscular prophylactic antibiotic injection, a parietal injection of xylocaine, the administration of painkillers, and a progressive reintroduction of diet.

One-anastomosis Gastric Bypass OAGB in Rats

This protocol is for the performance of one-anastomosis gastric bypass (OAGB) on rat. The operator performs a long and tubular-stapled gastric pouch followed by hand-sewn anastomosis. For this model, the operator reproduces the same ratio between biliopancreatic and common limb in humans; therefore, the biliopancreatic limb measures 35 cm.

The goal of this protocol is to set up a preclinical model of bariatric surgery and, more specifically, OAGB in obese rats. Based on this preclinical ...

Palatable Western-style Cafeteria Diet as a Reliable Method for Modeling Diet-induced Obesity in Rodents

Obesity is rapidly increasing in incidence in developed and developing countries and is known to induce or exacerbate many diseases.&#160;The health burden of obesity and its comorbid conditions highlight the need for better understanding of its pathogenesis, yet ethical constraints limit studies in humans. To this end externally valid models of obesity in laboratory animals are essential for the understanding of being overweight and obesity.&#160;While many species have been used to model the range of changes that accompany obesity in humans, rodents are most commonly used. Our laboratory has developed a western-style cafeteria diet that consistently leads to considerable weight gain and markers of metabolic disease in rodents. The diet exposes rodents to a variety of highly palatable foods to induce hyperphagia, modeling the modern western food environment. This diet rapidly induces weight gain and body fat accumulation in rats allowing for the study of effects of overeating and obesity. While the cafeteria diet may not provide the same control over macronutrient and micronutrient profile as purified high-fat or high-fat, high-sugar diets, the cafeteria diet typically induces a more severe metabolic phenotype than that observed with purified diets and is more in line with metabolic disturbances observed in the overweight and obese human population.

This protocol describes the use of a highly palatable, western-style cafeteria diet to model overeating and obesity in rodents. Here, we provide a detailed outline of food selection, preparation and measurement, and explain methodological factors that assist in generating a robust and reproducible phenotype.

Obesity is rapidly increasing in incidence in developed and developing countries and is known to induce or exacerbate many diseases.&#160;The health ...