利拉鲁肽联合骨髓间充质干细胞介导胃肠激素对链脲佐菌素诱导的1型糖尿病大鼠模型糖代谢的影响

Effect of liraglutide combined with bone marrow mesenchymal stem cells on glucose metabolism in experimental type 1 diabetic rats by regulating gastrointestinal hormones

目的探讨利拉鲁肽（liraglutide）与骨髓间充质干细胞（bonemarrowmesenchymalstemcells，BM．MSCs）联合治疗对1型糖尿病（T1DM）大鼠模型糖代谢的影响。方法以60mg／kg剂量腹腔注射链脲佐菌素制备T1DM大鼠模型，随后按随机数字表法分为（1）T1DM组（n=10）、（2）BM—MSCs治疗组（n=10）、（3）利拉鲁肽治疗组（LIRA，n=10）、（4）利拉鲁肽联合BM-MSCs治疗组（LIRA＋BM．MSCs，n=10），给予相应干预8周。测定各组大鼠随机血糖、HbA。24h尿量和24h尿蛋白；采用ELISA法检测血清胰岛素、c肽、胰升糖素、胃泌素、胆囊收缩素和胰升糖素样肽1（GLP-1）等激素水平：免疫组化测定胰腺组织中胰岛素和胰升糖素的表达。结果8周后，LIRA＋BM．MSCs组大鼠血糖、HbA1C、24h尿量和24h尿蛋白较其他3组均明显降低（P〈0．05）；分别与TIDM组和BM—MSCs组比较，LIRA＋BM—MSCs组血清胰岛素、c肽、胃泌素、胆囊收缩素和GLP-1水平均明显升高，胰升糖素水平降低（均P〈0．05）；与LIRA组比较，LIRA＋BM—MSCs组血清胰岛素、C肽、胃泌素和胆囊收缩素水平均明显升高（P〈0．05），胰升糖素和GLP-1水平无差异（P〉0．05）；相关分析发现，血清胰岛素水平分别与胃泌素（r=0．544，P〈0．01）、胆囊收缩素（r=0．710，P〈0．01）和GLP-1（r=0．669，P〈0．01）水平呈正相关，与胰升糖素水平呈负相关（r=-0．506，P〈0．01），胰升糖素与胃泌素（r=-0．364，P〈0．05）、胆囊收缩素（r=-0．433，P〈O．01）和GLP-1（r=-0．591，P〈0．01）呈负相关；免疫组化显示，LIRA＋BM．MSCs组大鼠胰腺组织胰岛素的阳性面积比例较其他3组均明显升高，胰升糖素比例降低（均P〈0．05）。结论利拉鲁肽联合骨髓间充质干细胞可能比两者单独治疗能够更好地介导胃肠激素分泌而改善TIDM大鼠的糖代谢。

Objective To investigate the effect of liraglutide combined with bone marrow mesenchymal stem cells （BM-MSCs） on glucose metabolism in experimental type 1 diabetic （T1DM） rats. Methods T1DM rats were established by injecting 60 mg/kg streptozotocin. According to random number table, they were divided into T1DM group （ n= 10）, BM-MSCs group （ n= 10）, LIRA group （ n = 10）, and LIRA＋BM-MSCs group （ n = 10）, and were treated for 8 weeks respectively. Random blood glucose, 24 h urine volume and protein excretion were tested. Serum concentrations of insulin, C-peptide, glucagon, gastrin, cholecystokinin, and glucagon-like peptid 1 （ GLP-1 ） were assayed by ELISA. Expressions of insulin and glucagon in pancreas were measured by immunohistochemistry. Results After 8 weeks, the levels of random blood glucose, HbA1c, 24 h urine volume and 24 h urinary protein excretion in group 4 were significantly decreased compared to the other three groups （P〈0. 05 ）. Compared to T1DM group and BM-MSCs group, the levels of insulin, C-peptide, gastrin, cholecystokinin and GLP-1 in LIRA＋BM-MSCs group were significantly increased, while glucagon was decreased （ P〈0.05 ）. Compared to group LIRA, the levels of insulin, C-peptide, gastrin, and cholecystokinin in LIRA ＋ BM-MSCs group were increased （P 〈 0. 05 ）, there was no significantly difference in glucagon or GLP-1 （ P〉0.05 ）. The analysis revealed that the level of insulin was positively correlated with gastrin （ r = 0. 544, P〈0. 01 ）, eholeeystokinin （ r = 0. 710, P〈0. 01 ） and GLP-1 （ r = 0. 669, P〈0.01 ）, but was negatively correlated with glucagon （r =- 0. 506, P〈0.01 ） ; the level of glucagon was negatively correlated with gastrin （ r = -0. 364, P〈0.05 ）, cholecystokinin（ r = -0. 433, P〈0.01 ） and GLP-1 （ r = -0. 591, P〈 0.01 ）. Compared to the other three groups respectively, immunohistochemistry displayed that the positive area of insulin in pancreas was significantly increased in LIRA＋BM-MSCs group, while that of glucagon was decreased （ P〈 0.05）. Conclusions By means of regulating gastrointestinal hormones efficiently, combination of liraglutide with BM-MSCs may improve glucose metabolism more efficaciously than treatment with a single agent in T1DM rats.