降糖三黄片改善2型糖尿病小鼠胰岛β细胞炎症反应实验研究

Experiment Study of Jiangtang Sanhuang Tablets on Improving Inflammatory Responses in Pancreatic β Cells of Mice with Type 2 Diabetes Mellitus

目的:研究降糖三黄片通过减少db/db小鼠胰岛β细胞炎症反应改善胰岛β细胞功能的机制。方法:将雄性6周龄db/db小鼠24只随机平均分为3组:模型组、降糖三黄片组、二甲双胍组;同窝别db/m小鼠8只设为对照组。降糖三黄片组予降糖三黄片蒸馏水溶液灌胃,2.64 g/kg,每天1次;二甲双胍组予盐酸二甲双胍蒸馏水溶液灌胃,0.2 g/kg,每天1次;对照组及模型组均予等量蒸馏水灌胃,每次1 mL,每天1次。每2周测体质量及血糖,干预6周后取材。摘眼球取血测血糖及胰岛素,取血清用酶联免疫吸附测定(ELISA)法测白细胞介素(IL)-1β、肿瘤坏死因子-α(TNF-α)及细胞间黏附分子-1(ICAM-1)的含量;取胰腺行病理片HE染色。结果:①空腹血糖(FBG):干预后,降糖三黄片组及二甲双胍组FBG干预前后比较,差异有统计学意义(P<0.05);降糖三黄片组FBG与二甲双胍组比较,差异无统计学意义(P>0.05)。②体质量:干预后,二甲双胍组小鼠体质量上升(P<0.05)。③胰岛素分泌指标:与模型组比较,降糖三黄片组空腹胰岛素(FINS)明显升高,差异有统计学意义(P<0.05);降糖三黄片组FINS与二甲双胍组比较,差异无统计学意义(P>0.05)。与模型组比较,降糖三黄片组、二甲双胍组胰岛β细胞分泌指数(HOMA2%β)均升高,差异均有统计学意义(P<0.05);降糖三黄片组HOMA2%β与二甲双胍组比较,差异无统计学意义(P>0.05)。④炎症因子指标:与对照组比较,模型组IL-1β、TNF-α及ICAM-1水平均上升(P<0.05);与模型组比较,降糖三黄片组及二甲双胍组IL-1β、TNF-α及ICAM-1水平均降低(P<0.05),降糖三黄片组IL-1β、TNF-α及ICAM-1水平分别与二甲双胍组比较,差异均无统计学意义(P>0.05)。⑤胰腺病理结果:与对照组比较,模型组小鼠胰岛萎缩,部分坏死,结构不完整。与模型组比较,降糖三黄片组胰岛萎缩情况较轻,胰腺炎症较轻,结构相对完整。与二甲双胍组比较,降糖三黄片组胰岛β细胞数量增多,界限较清楚。与对照组比较,降糖三黄片组胰岛细胞排列拥挤,胞浆不丰富,细胞核较密集。结论:降糖三黄片可能通过减少IL-1β、TNF-α及ICAM-1表达,改善db/db小鼠胰岛β细胞的炎症反应,保护胰岛β细胞功能。

Objective:To study the mechanism of Jiangtang Sanhuang tablets on improving function of pancreaticβcells by reducing inflammatory responses in pancreaticβcells in db/db mice.Methods:A total of 24 male 6-week-old db/db mice were randomly and equally divided into three groups,namely the model group,the Jiangtang Sanhuang tablets group and the metformin group;eight db/m mice in the same litter were set as the control group.The Jiangtang Sanhuang tablets group was given gastric gavage with distilled water solution of Jiangtang Sanhuang tablets with 2.64 g/kg,once a day.The metformin group was given gastric gavage with distilled water solution of metformin hydrochloride with 0.2 g/kg once a day.The control group and the model group were given distilled water with equivalent volume,with 1mL each time and once a day.The body weight and blood glucose were measured every two weeks,and the materials were taken after six weeks of intervention.Eyeballs were removed to collect blood to measure blood glucose and insulin,and serum were taken to test the contents of interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α)and intercellular adhesion molecule-1(ICAM-1)by ELISA method;pancreas was extracted to perform HE staining of pathological slices.Results:①Fasting blood glucose(FBG):After intervention,there was a significant difference being found in the comparison of FBG in the Jiangtang Sanhuang tablets group and the metformin group before and after intervention(P<0.05);there was no significant difference being found in the comparison of FBG between the Jiangtang Sanhuang tablets group and the metformin group(P>0.05).②Body weight:After intervention,the body weight of mice in the metformin group was increased(P<0.05).③Insulin secretion indexes:The level of fasting insulin(FINS)in the Jiangtang Sanhuang tablets group was significantly increased when compared with that in the model group,the difference being significant(P<0.05);there was no significant difference being found in the comparison of level of FINS between the Jiangtang Sanhuang tablets group and the metformin group.(P>0.05)The levels of HOMA2%βin the Jiangtang Sanhuang tablets group and the metformin group were increased when respectively compared with that in the model group,differences being significant(P<0.05);there was no significant difference being found in the comparison of level of HOMA2%βbetween the Jiangtang Sanhuang tablets group and the metformin group(P>0.05).④Inflammatory factors indexes:The levels of IL-1β,TNF-αand ICAM-1 in the model group were increased when compared with those in the control group(P<0.05);the above levels in the Jiangtang Sanhuang tablets group and the metformin group were decreased when compared with those in the model group(P<0.05),and there was no significant difference being found in the comparison of the above levels between the Jiangtang Sanhuang tablets group and the metformin group(P>0.05).⑤Pathologic findings of pancreas:The atrophy,partial necrosis and incomplete structure of pancreatic islets were found in the mice in the model group when compared with those in the control group.The atrophy of pancreatic islets and pancreas inflammatory in the Jiangtang Sanhuang tablets group were relatively mild,and the structure of pancreatic islets were relatively complete when compared with those in the model group.The number of pancreatic isletβcells in the Jiangtang Sanhuang tablets group was increased and the boundaries were clearer when compared with those in the metformin group.The cell alignment of pancreatic islets in the Jiangtang Sanhuang tablets group was crowded,the cytosolic was not abundant,and the nucleus was relatively dense when compared with those in the control group.Conclusion:Jiangtang Sanhuang tablets can improve the inflammatory responses in pancreaticβcells of db/db mice and protect function of pancreaticβcell by reducing expressions of IL-1β,TNF-αand ICAM-1.