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. Type 2 diabetes in adolescents and young adults. Lancet Diabetes Endocrinol. 6 (1), 69-80 (2018).</li>
	<li>Song, Y., Zhao, F. Bibliometric analysis of metabolic surgery for type 2 diabetes: current status and future prospects. Updates Surg. 74 (2), 697-707 (2022).</li>
	<li>Cummings, D. E., Rubino, F. Metabolic surgery for the treatment of type 2 diabetes in obese individuals. Diabetologia. 61 (2), 257-264 (2018).</li>
	<li>Buchwald, H. The evolution of metabolic/bariatric surgery. Obes Surg. 24 (8), 1126-1135 (2014).</li>
	<li>Akalestou, E., Miras, A. D., Rutter, G. A., le Roux, C. W. Mechanisms of weight loss after obesity surgery. Endocr Rev. 43 (1), 19-34 (2022).</li>
	<li>Rubino, F. et al. The mechanism of diabetes control after gastrointestinal bypass surgery reveals a role of the proximal small intestine in the pathophysiology of type 2 diabetes. Ann Surg. 244 (5), 741-749 (2006).</li>
	<li>Rubino, F., Marescaux, J. Effect of duodenal-jejunal exclusion in a non-obese animal model of type 2 diabetes: a new perspective for an old disease. Ann Surg. 239 (1), 1-11 (2004).</li>
	<li>Akalestou, E., et al. Establishing a successful rat model of duodenal- jejunal bypass: A detailed guide. Lab Anim. 53 (4), 362-371 (2019).</li>
	<li>Yu, H. H., Hsieh, M. C., Wu, S. Y., Sy, E. D., Shan, Y. S. Effects of duodenal-jejunal bypass surgery in ameliorating nonalcoholic steatohepatitis in diet-induced obese rats. Diabetes Metab Syndr Obes. 12, 149-159 (2019).</li>
	<li>Garciacaballero, M., Navarrete, S., Favretti, F., Celik, A., Del Castillo, D. Diabetes surgery in type 2 BMI 24-29 vs IMC 30-34 diabetic patients: is there differences among restrictive, malabsorptive and gastric bypass procedures? Nutr Hosp. 28 Suppl 2:23-30 (2013).</li>
	<li>Vidal, J., Jim&#233;nez, A. Diabetes remission following metabolic surgery: is GLP-1 the culprit? Curr Atheroscler Rep. 15 (10):357 (2013).</li>
	<li>Speck, M., Cho, Y. M., Asadi, A., Rubino, F., Kieffer, T. J. Duodenal-jejunal bypass protects GK rats from {beta}-cell loss and aggravation of hyperglycemia and increases enteroendocrine cells coexpressing GIP and GLP-1. Am J Physiol Endocrinol Metab. 300 (5), E923-932 (2011).</li>
	<li>Ashrafian, H. et al. Metabolic surgery: an evolution through bariatric animal models. Obes Rev. 11(12), 907-920 (2010).</li>
	<li>Rao, R. S., Rao, V., Kini, S. Animal models in bariatric surgery--a review of the surgical techniques and postsurgical physiology. Obes Surg. 20 (9), 1293-1305 (2010).</li>
	<li>Im, Y. R. et al. A systematic review of animal models of NAFLD finds high-fat, high-fructose diets most closely resemble human NAFLD. Hepatology. 74 (4), 1884-1901 (2021).</li>
	<li>Kleinert, M. et al. Animal models of obesity and diabetes mellitus. Nat Rev Endocrinol. 14 (3), 140-162 (2018).</li>
	<li>Furman, B. L. Streptozotocin-induced diabetic models in mice and rats. Curr Protoc. 1 (4):e78 (2021).</li>
	<li>Buchwald, H., Varco, R. L., Moore, R. B., Schwartz, M. Z. Intestinal bypass procedures. Partial ileal bypass for hyperlipidemia and jejunoileal bypass for obesity. Curr Probl Surg. 1-51 (1975).</li>
	<li>DeMaria, E. J. Bariatric surgery for morbid obesity. N Engl J Med. May 24;356(21):2176-2183 (2007).</li>
	<li>Bolling, C. F., Armstrong, S. C., Reichard, K. W., Michalsky, M. P. Metabolic and bariatric surgery for pediatric patients with severe obesity. Pediatrics. 144 (6), e20193224 (2019).</li>
	<li>Mason, E. E. Gastric bypass for morbid obesity. Surg Annu. 11, 99-126 (1979).</li>
	<li>Cheng, Y. et al. Sleeve gastrectomy with bypass of proximal small intestine provides better diabetes control than sleeve gastrectomy alone under postoperative high-fat diet. Obes Surg. 29 (1), 84-92 (2019).</li>
	<li>Rubino, F. et al. The role of the small bowel in the regulation of circulating ghrelin levels and food intake in the obese Zucker rat. Endocrinology. 146 (4), 1745-1751 (2005).</li>
	<li>Zachariah, P. J. et al. 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. Obes Surg. 26 (12), 2862-2872 (2016).</li>
	<li>Jiang, B., et al. Role of proximal intestinal glucose sensing and metabolism in the blood glucose control in type 2 diabetic rats after duodenal jejunal bypass surgery. Obes Surg. 32 (4), 1119-1129 (2022).</li>
	<li>Han, H. F. et al. Duodenal-jejunal bypass increases intraduodenal bile acids and upregulates duodenal SIRT1 expression in high-fat diet and streptozotocin-induced diabetic rats. World J Gastroenterol. 28 (31), 4338-4350 (2022).</li>
	<li>Yan, S., et al. Reduction of intestinal electrogenic glucose absorption after duodenojejunal bypass in a mouse model. Obes Surg. 23 (9), 1361-1369 (2013).</li>
	<li>Arble, D. M. et al. Metabolic comparison of one-anastomosis gastric bypass, single-anastomosis duodenal-switch, Roux-en-Y gastric bypass, and vertical sleeve gastrectomy in rat. Surg Obes Relat Dis. 14 (12), 1857-1867 (2018).</li>
	<li>Liang, Y. et al. Duodenal-jejunal bypass surgery reverses diabetic phenotype and reduces obesity in db/db Mice. Curr Chem Genom Transl Med. 11, 41-49 (2017).</li>
	<li>Barataud, A. et al. Absence of role of dietary protein sensing in the metabolic benefits of duodenal-jejunal bypass in the mouse. Sci Rep. 7, 44856 (2017).</li>
	<li>Close-up on lab animal microsurgery. Lab Anim (NY). 35(1), 43 (2006).</li>
	<li>Couceiro, J., Castro, R., Tien, H., Ozyurekoglu, T. Step by step: microsurgical training method combining two nonliving animal models. J Vis Exp. (99), e52625 (2015).</li>
	<li>Nasser, K. M., Wahba, H. A., Kamal, E., El-Makhzangy, A. M., Bahaa, N. Animal model for training and improvement of the surgical skills in endolaryngeal microsurgery. 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.
<p clas...

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.

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 ...

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

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 ...

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. ...

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 ...

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 ...

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 ...

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 ...

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 ...

One-anastomosis Gastric Bypass (OAGB) in Rats

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 ...

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 ...