Name: sleeve gastrectomy in mice using surgical clips
Uploaded: 2020-11-14T13:00:00
Description: Watch this Scientific Journal Video about Sleeve Gastrectomy in Mice using Surgical Clips at JoVE.com

We thank Miss Isabelle Lu for the English narration.

All procedures for animal use were approved by the National Yang-Ming University Institutional Animal Care and Use Committee and complied with the &#8220;Guide for the Care and Use of Laboratory Animals. 8th edition, 2011&#8221;. The anesthetic procedure was performed according to the guideline for the pain control, anesthesia, pre-operative, and postoperative care for the experimental animals from National Yang Ming University, ICAUC-016, 2015, 1st edition.
1. Animal prepa...

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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+ameliorates+cardiovascular+risk+in+obese+diabetic+patients:+Influence+of+different+surgical+procedures.">Metabolic surgery ameliorates cardiovascular risk in obese diabetic patients: Influence of different surgical procedures.</a> Surgery for Obesity and Related Diseases. 14 (12), 1832-1840 (2018).</li><li>Lee, W. J., Aung, L. <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+for+Type+2+Diabetes+Mellitus:+Experience+from+Asia.">Metabolic Surgery for Type 2 Diabetes Mellitus: Experience from Asia.</a> Diabetes &amp; Metabolism. 40 (6), 433-443 (2016).</li><li>Yin, D. 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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=Effects+of+sleeve+gastrectomy+in+high+fat+diet-induced+obese+mice:+respective+role+of+reduced+caloric+intake,+white+adipose+tissue+inflammation+and+changes+in+adipose+tissue+and+ectopic+fat+depots.">Effects of sleeve gastrectomy in high fat diet-induced obese mice: respective role of reduced caloric intake, white adipose tissue inflammation and changes in adipose tissue and ectopic fat depots.</a> Surgical Endoscopy. 28 (2), 592-602 (2014).</li><li>Rubino, F., R'Bibo, S. L., del Genio, F., Mazumdar, M., McGraw, T. 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Most of the prospective cohort studies have confirmed that increasing the body mass index is related to increasing the mortality. Obesity is associated with a higher risk of diabetes, cardiovascular diseases (CVD), and death in both sexes. The specific pathologic processes have established a link between diabetes, obesity, and mortality14. The systematic reviews indicate the benefits of bariatric surgery in reducing risk factors for CVD with evidence for ventricular hypertrophy regression and impr...

The prevalence of obesity has increased nearly threefold worldwide since 1975. In 2016, more than 1.9 billion adults older than 18 years were overweight and over 650 million adults were obese. The prevalence of T2D in the adult population has also doubled from 4.7% to 8.5% with the number rising from 108 million to 422 million adults between 1980 and 2014 globally1. Most bariatric surgeries result in significant weight loss, a reduction of body fat, and improvement in glucose tolerance, insulin resistance and diabetes control2. In addition to weight loss and remission of T2D, bariatric surgery further produces additional health advantages such as hypertension control and a lower incidence of certain types of obesity related cancer development and progression3. Bariatric surgery also induces durable complete, partial remission of T2D, and decreases the risk of ten-year major coronary heart disease (CHD) and some degree of cerebrovascular risk. However, the underlying mechanisms are not completely understood4.
There are several procedures for bariatric/metabolic surgery. Both restrictive and gastrointestinal bypass surgeries provide positive effects on metabolism5. Of the bariatric surgical models, SG and modified Roux-en-Y gastric bypass have higher success and lower mortality rates and demonstrate reliable restrictive and gastrointestinal bypass surgery models in mice6. Although the gastrointestinal bypass surgery provides more weight loss and a significant improvement of glucose tolerance and liver steatosis than the restrictive procedure in the long term, the SG still produces good control of body weight and glucose levels and is easy to perform with a low incidence of complications6. The proportion of SG increased from 30% to 54% and Roux-en-Y gastric bypass surgery decreased from 52% to 32% from 2008 to 20147. Currently, laparoscopic SG is the most commonly performed bariatric procedure at the national level within academic centers in the United States8. Although there have been many published reports regarding the pathophysiological processes between bariatric surgery, diabetes and obesity, we need more animal experiments to further explore unknown mechanisms.
This protocol aims to produce an animal method to investigate the mechanisms underlying the outcomes seen after bariatric surgery in humans. The current translational study may provide insights on the mechanism of obesity and T2D treatment from the SG.

<table>
	<tbody>
		<tr>
			<td>0.9% normal saline</td>
			<td>China Chemical &#38; Pharmaceutical Co., Ltd.</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>5-0 monofilament suture</td>
			<td>Shineteh Inc. Taiwan</td>
			<td></td>
			<td>Ethicon</td>
		</tr>
		<tr>
			<td>Alm</td>
			<td>Shineteh Inc. Taiwan</td>
			<td>ST-A072PK</td>
			<td>UNIK</td>
		</tr>
		<tr>
			<td>Animal Anesthesia</td>
			<td>Step Technology</td>
			<td>0712VAP11076</td>
			<td>Matrx</td>
		</tr>
		<tr>
			<td>Barraqer-Troutman forceps</td>
			<td>Shineteh Inc. Taiwan</td>
			<td>ST-N309(H-4410)</td>
			<td>UNIK</td>
		</tr>
		<tr>
			<td>cefazolin</td>
			<td>Taiwan veteran pharmacy company</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Chow diet</td>
			<td>Research Diets</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Glucose strips and glucometer</td>
			<td>BeneCheck</td>
			<td>BKM13-1</td>
			<td>BeneCheck</td>
		</tr>
		<tr>
			<td>HbA1c kit</td>
			<td>Level</td>
			<td>OKEH00661</td>
			<td></td>
		</tr>
		<tr>
			<td>Hematology system</td>
			<td>Fuji</td>
			<td>FUJI DRI-CHEM 4000i</td>
			<td>Europe</td>
		</tr>
		<tr>
			<td>High Fat diet</td>
			<td>Research Diets</td>
			<td>1810724</td>
			<td>LabDiet</td>
		</tr>
		<tr>
			<td>Iris Forceps</td>
			<td>Shineteh Inc. Taiwan</td>
			<td>ST-1210</td>
			<td>UNIK</td>
		</tr>
		<tr>
			<td>Iris Scissors</td>
			<td>Shineteh Inc. Taiwan</td>
			<td>ST-S011</td>
			<td>UNIK</td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>Panion &#38; BF biotech INC</td>
			<td>8547</td>
			<td>Panion &#38; BF</td>
		</tr>
		<tr>
			<td>Ketoprofen</td>
			<td>Sigma</td>
			<td>22071154</td>
			<td>Sigma-Aldrich</td>
		</tr>
		<tr>
			<td>Stereo microscope</td>
			<td>MicroTech</td>
			<td>SZ-5T</td>
			<td>MicroTech</td>
		</tr>
		<tr>
			<td>Surgical clip (M)</td>
			<td>Echicon Inc., Somerville, NJ</td>
			<td></td>
			<td>Size M, 5mm</td>
		</tr>
		<tr>
			<td>Vidisic gel</td>
			<td>Dr. Gerhard Mann Chem.-pharm. Fabrik GmbH</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

sleeve gastrectomy in mice using surgical clips

The number of people who are overweight and obese is continually increasing both in the adult and adolescent populations. This coincides with the increased universal phenomenon of type 2 diabetes (T2D) and other metabolic problems. Bariatric surgery, such as SG, is currently one of the most effective and commonly used long-term treatment for obesity and T2D, but the association between them is not completely explored yet. The mechanisms underlying the outcomes seen after bariatric surgery in humans can be investigated based on preclinical animal studies. The SG reduces body weight, glucose levels and many metabolic parameters, and is easy to perform with a low incidence of complications. The goal of this work is to provide a simple method and an uncomplicated preclinical model of bariatric surgery in animals for researchers.

Results of the operation are shown in Figure 3 and Figure 4. The survival rate of the study was 90%. One mouse died in the sham group because of weakness and another mouse died in the SG group on the third day after operation for unknown reasons. Results obtained on weight loss (Figure 3A) after operation (at week 3) on HFD fed mice were quite similar to those observed in humans, with a final body weight loss of about 15-20%<sup cla...

Watch this Scientific Journal Video about Sleeve Gastrectomy in Mice using Surgical Clips at JoVE.com

Sleeve Gastrectomy in Mice using Surgical Clips

The prevalence of diabetes and obesity is continuously increasing worldwide. The mechanisms between diabetes, obesity and their associated mortality and co-morbidities need to be further investigated. Here, we present a protocol for sleeve gastrectomy (SG) in animals as an uncomplicated preclinical model of bariatric surgery.

The number of people who are overweight and obese is continually increasing both in the adult and adolescent populations. This coincides with the ...

The chief target of the surgical intervention is to measure the metabolic effects and the insulin resistance of sleeve gastrectomy in a more assessable mouse model. This protocol aims to produce an animal model for the research of the mechanisms that underline the results seen after a bariatric surgery in humans, which may provide insights into the pathogenesis of obesity and type 2 diabetes treatment. For sleeve gastrectomy and sham operation, after confirming the appropriate depth of anesthesia by toe stimulation, apply the Carbomer 0.2%eye gel on the mouse's eyes to prevent dry eyes.The mouse is placed and fixed on the surgical table. Remove and clean the remaining hair with a depilatory cream. Disinfect the abdomen of the mouse with povidone-iodine solutions alternating with alcohol for three applications.Perform the SG procedure with the aid of a magnifying dissecting microscope or magnifier as necessary to prevent any unpredictable bleeding and hypovolemic shock. Make a one to 1.5 centimeters in length midline incision at the upper abdomen. Use two curved smooth serrated micro-forceps to gently move the stomach and complete externalize it.Move the intestine to the bare skin on the side and cover it with warm saline moistened gauze to prevent dehydration. Carefully divide the gastrosplenic ligament connecting the left stomach to the spleen using a cotton-tipped probe and electrocautery as needed, thereby dissecting the gastric fundus from the surrounding spleen and other internal organs. Stretch the fundus and pylorus of the stomach gently and laterally with forceps, identify the midline of the stomach and carefully apply two surgical clips using the LIGACLIP applier to the 15%medial segment of the stomach from the gastroesophageal junction inferiorly and pylorus superiorly.Clamp and isolate around 75-80%of stomach, thereby creating the entire lateral sleeve of the stomach. Resect the lateral isolated stomach with a microscissors. Use cotton swabs to remove any residual food from the stomach and then clean the cut edge of the stomach with povidone-iodine aqueous solution.Oversew the clip line with a 5-0 monofilament nonabsorbable suture to make sure of no leakage. Knot the ends of the suture and anchor to the clips on either end. Return the stomach and intestine to the proper position in the abdominal cavity and close the abdomen with a running 5-0 monofilament nonabsorbable suture to the abdominal wall.For sham procedure of SG, perform a similar procedure as described previously with a midline laparotomy, externalize the intestine and stomach using 37 degree Celsius wet warm saline gauze coverage for five minutes. Return the stomach and intestine to the proper sites of these internal organs. Administer ketoprofen two to five milligrams per kilogram and cefazolin 25 milligrams per kilogram subcutaneously or intraperitoneally after surgery for the pain control and infection prevention.Stop the isoflurane and continuing with a room air flow of three to five liters per minute until the mouse is fully awake. Keep watching the mouse while it regains mobility and begins to walk around the cage. Place the mouse in the independent incubator around 30 degrees Celsius in temperature condition for five days, making sure there is only one mouse per cage.In our experienced hands, the successful rate of SG was around 90%after the learning period. The body weight of sham operated and high-fat diet fed mice gained steadily. But the sleeve gastrectomy and high-fat diet fed mice decreased continuously.The difference of the cholesterol level was significant between the high-fat diet fed mice with sham and sleeve gastrectomy. This might indicate that oxidative stress improvement after the sleeve gastrectomy. The difference of the levels of hemoglobin A1C and IPGTT were significant as well.Between the high-fat diet fed mice with sham and sleeve gastrectomy and by indicating insulin-resistance and sugar control improvement after the sleeve gastrectomy. This animal model can help us to investigate the clinical relevance of diabetes and obesity in using a brief surgical procedure with sleeve gastrectomy. This procedure can shorten the operation time and improve the surgical survival rate.The main advantage of this study is to imitate the clinical outcomes of bariatric surgery in human patients using a simple surgical mouse model.

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