国产胶原酶在小鼠胰岛分离中的效果研究  

Study on the effectiveness of a domestic collagenase in mouse islet isolation

作　　者：宋佳文 徐福远 蒋煊 吴国娣 蒋必颖 傅红兴 SONG Jia-wen;XU Fu-yuan;JIANG Xuan;WU Guo-di;JIANG Bi-ying;FU Hong-xing(Department of Pharmacy,Lishui People’s Hospital,Lishui 323000,China;School of Pharmaceutical Sciences,Wenzhou Medical University,Wenzhou 325035,China)

机构地区：[1]丽水市人民医院药学部,浙江丽水323000 [2]温州医科大学药学院,浙江温州325035

出　　处：《肝胆胰外科杂志》2019年第10期616-619,共4页Journal of Hepatopancreatobiliary Surgery

基　　金：浙江省重点研发计划项目（2018C03013）;温州市科技计划项目（Y20160079）;国家大学生创业实践项目（201810343048S）

摘　　要：目的研究一种国产胶原酶在小鼠胰岛分离中的分离效果,探索该胶原酶在胰岛分离中应用的可行性。方法将国产胶原酶分别配成不同浓度的胶原酶溶液和含中性蛋白酶的胶原酶的溶液,经胰管逆行灌注胶原酶的方法进行小鼠胰岛分离,采用Ficoll液对胰岛进行纯化,计数所得的胰岛细胞团,培养6h后检测活性,对最佳分离结果的胰岛做同系糖尿病小鼠的肾被膜下移植,监测术后血糖和进行组织学检查,以进口Sigma胶原酶V为对照。结果国产胶原酶在小鼠胰腺消化时间、所得胰岛数量和活性效果上跟进口胶原酶V有一定差距(P<0.01),但含中性蛋白酶的国产胶原酶组在小鼠胰岛分离数量和当量上与进口胶原酶组无差异(P>0.05),分离的胰岛在体内移植后降血糖效果与进口胶原酶组无差异(P>0.05)。结论国产胶原酶结合中性蛋白酶可用于小鼠胰岛的分离。Objective To study the effectiveness of a domestic collagenase in mouse islet isolation, and explore the feasibility of its application in islet isolation. Methods The domestic collagenase was prepared into collagenase solution and collagenase solution containing neutral protease with different concentration. The islet was isolated by retrograde infusion of collagenase into pancreatic duct, and purified by Ficoll solution. After cultured for 6 hours, the viability of islets was detected. The islets isolated from the optimal condition were transplanted under the renal capsule of the homologous diabetic mice. And the blood glucose, histological examinations were also monitored after transplantation. The imported Sigma collagenase V was used as a control. Results The domestic collagenase had a certain gap with Sigma collagenase V in pancreas digestion time, the number of islets obtained and the viability of islets (P<0.01). But for the group of domestic collagenase containingneutral protease, there was no significant difference from those of Sigma collagenase V group in terms of the number, equivalent of mouse islets obtained, or the blood glucose control after transplantation in vivo (P>0.05). Conclusion The domestic collagenase containing neutral protease can be used for the isolation of mouse islets.

关 键 词：国产胶原酶 中性蛋白酶 胰岛分离 胰岛移植 小鼠 

分 类 号：R587[医药卫生—内分泌]