胃浆膜下层与门静脉胰岛移植治疗糖尿病大鼠的实验研究

Experimental study on gastric subserosa and portal vein islet transplantation for treatment of rat diabetes

目的通过比较胃浆膜下层与门静脉胰岛移植治疗糖尿病大鼠的功效,探讨胰岛移植的最适宜部位。方法链脲佐菌素诱导糖尿病大鼠,将其分为胃浆膜组、门静脉组、对照组,每组6只;经胆总管灌注胶原酶Ⅺ消化胰腺组织,分离、纯化胰岛,采用双硫腙(DTZ)特性染色并计数胰岛当量(IEQ)及纯度,台盼蓝染色判断胰岛活性;同期分别将500 IEQ大鼠胰岛移植入胃浆膜下层和门静脉,对照组输注等量RPMI1640液体,移植后分别检测大鼠的血糖浓度,腹腔葡萄糖耐量实验,评判胰岛功能。结果每只大鼠可获取胰岛(310±42)个,DTZ染色显示胰岛纯度为(89.00±4.52)%;台盼蓝染色显示胰岛活性良好[(89.08±3.76)%];胰岛移植后胃浆膜组、门静脉组大鼠血糖较之前下降(P<0.05),与对照组血糖浓度比较,差异有统计学意义(P<0.05);胃浆膜组与门静脉组胰岛移植后大鼠血糖浓度比较,差异有统计学意义(P<0.05);胃浆膜组与门静脉组大鼠有效控制血糖时间分别为(18.0±1.9)和(11.6±2.5)d,差异有统计学意义(P<0.05)。胃浆膜组与门静脉组大鼠腹腔葡萄糖耐量实验提示糖耐受功能良好。结论短期内,胃浆膜下层胰岛移植治疗糖尿病大鼠的疗效优于门静脉移植;与门静脉相比,胃浆膜下层胰岛移植操作简单,安全性高,是胰岛移植的最适宜部位之一。

Objective To explore the most suitable transplant site for islet transplantation by comparing the efficacy of gastric subserosal transplantation and portal vein islet transplantation in the treatment of diabetic rats.Methods Streptozotocin-induced diabetic rats were divided into gastric subserous group(n=6),portal vein group(n=6)and control group(n=6).Type XI collagenase was injected through choledoch to digest the pancreas,and the islets were isolated and purified.The specificity of the islet was judged by dithizone(DTZ)staining,and IEQ and islets purity were counted.The activity of islets was determined by trypan blue test.500IEQ fresh islets were transplanted into the gastric subserosa and into the portal vein,respectively.The control group was infused with the same amount of RPMI 1640.After islet grafting in the diabetic rats,the changes in blood glucose and intraperitoneal glucose tolerance were evaluated to assess the function of islets.Results The number of islets was(310±42)in each transplanted rat,and the purity of islets was(89±4.52)%obtained by DTZ staining and the activity of isletswas(89.08±3.76)%obtained by trypan blue staining.The glucose levels in the gastric subserous and portal vein groups were significantly decreased after transplantation compared with those before transplantation(P<0.05)and significantly different from that in the control group(P<0.05).After transplantation the average blood glucose level of the gastric subserous group was significantly lower than that of the portal vein group(P<0.05).The time of effective control of blood glucose in the gastric subserous group and the portal vein group was(18.0±1.9)d and(11.6±2.5)d(P<0.05).The rats of the gastric subserous and portal vein groups had good intraperitoneal glucose tolerance.Conclusions In the short term,the effect of gastric subserous islet transplantation for treatment of rat diabetes is better than that of portal vein islet transplantation.Compared with portal vein islet transplantation,the operation of gastric subserous pancreatic islet transplantation is simple and safe,and gastric subserosa can be considered as one of the most suitable parts of islet transplantation.