摘要
目的:以自发性2型糖尿病GK大鼠为研究对象,应用全基因组表达谱芯片技术,旨在探讨参芪复方调节胰岛细胞功能的分子机制,为中医药防治2型糖尿病提供理论依据。方法:GK大鼠每日给予高脂饲料喂养,连续4周,随机从GK大鼠中选取大鼠检测随机血糖,验证2型糖尿病模型成功。实验分为4组,空白组,模型组,参芪复方组(1.44 g?kg-1),西格列汀组(16 mg·kg-1),于灌胃前、灌胃4周、灌胃8周测早上8:00各组大鼠尾尖血糖;灌胃8周后,运用酶联免疫吸附测定(ELISA)检测大鼠血清胰岛素(INS)水平;采用原位末端标记(TUNEL)荧光法定量检测并观察胰岛β细胞凋亡情况;运用全基因组表达谱芯片技术对各组大鼠胰腺组织进行差异基因检测;采用实时荧光定量聚合酶链式反应(Real-time PCR)检测关键差异基因的mRNA转录水平。结果:与空白组比较,灌胃前、灌胃4周、灌胃8周,GK大鼠各组血糖显著升高(P<0.01);灌胃4,8周,与模型组比较,各药物干预组大鼠血糖均显著较低(P<0.01)。灌胃8周,与空白组比较,模型组大鼠INS水平显著较低(P<0.01);与模型组比较,参芪复方组具有INS水平更高的趋势,西格列汀组INS水平显著较高(P<0.01)。灌胃8周,与空白组比较,模型组大鼠胰岛β细胞凋亡数明显升高(P<0.05);与模型组比较,参芪复方组、西格列汀组大鼠胰岛β细胞凋亡数均明显较低(P<0.05,P<0.01)。基因芯片及Real-time PCR检测均显示磷脂酰肌醇3-激酶受体1(PIK3R1)在参芪复方组/模型组中表达上调,在西格列汀组/模型组、模型组/空白组中表达下调;蛋白激酶B1(Akt1)在参芪复方组/模型组、西格列汀组/模型组中表达上调,在模型组/空白组中表达下调。结论:参芪复方可减少胰岛β细胞凋亡,增加胰岛素分泌,上调基因PIK3R1,Akt1的表达,推测具有养阴益气活血功效的参芪复方可能通过上调基因PIK3R1,Akt1的表达,抑制胰岛β细胞凋亡,改善胰岛β细胞功能,调节胰岛素分泌,从而防治2型糖尿病。
Objective:To explore the effect of Shenqi compound on isletβ-cell function in type 2 diabetic GK rats.The whole genome expression profile chip technology is used to explore the molecular mechanism of Shenqi compound regulating pancreatic islet cell function and provide theoretical basis for the prevention and treatment of type 2 diabetes with traditional Chinese medicine.Method:GK rats were fed with high-fat diet daily for 4 weeks.Rats were randomly selected from GK rats to detect random blood glucose and verified the success of type 2 diabetes model.Rats were divided into 4 groups,Wistar group,model group,Shenqi compound(1.44 g?kg-1)group and west glenn(16 mg?kg-1)group.After 8 weeks of gavage,the serum insulin(INS)levels were detected by enzyme-linked immunosorbent assay(ELISA).The apoptosis of isletβcells was detected by terminal-deoxynucleoitidyl transferase mediated nick end labeling(TUNEL)fluorescence method.Differential gene detection uses whole-genome expression profiling chip technology in each group of rat pancreatic tissues,the mRNA transcription level of key differential genes is detected by Real-time fluorescent quantitative polymerase chain reaction(Real-time PCR).Result:Compared with blank group,before gavage,4 weeks,8 weeks,GK rats have higher blood sugar in each group(P<0.01).Gavage for 4 weeks and gavage for 8 weeks,compared with model group,the blood sugar of rats in each drug intervention group was lower(P<0.01).Gavage for 8 weeks,compared with blank group,the INS level of model group was lower(P<0.01).Compared with model group,the Shenqi compound group had a higher INS level,and the sitagliptin group had a higher INS level(P<0.01).After gavage for 8 weeks,compared with the blank group,the number of pancreatic isletβ-cell apoptosis in the model group was higher(P<0.05).Compared with model group,the number of pancreatic isletβcell apoptosis in the Shenqi compound group and sitagliptin group was lower(P<0.05,P<0.01).Gene chip and Real-time PCR tests both showed that phosphatidylinositol 3-kinase receptor 1(PIK3R1)was up-regulated in the Shenqi compound group/model group,and down-regulated in the sitagliptin group/model group,model group/blank group.Protein kinase B1(Akt1)was expressed in the Shenqi compound group/model The expression was up-regulated in the group,sitagliptin group/model group,and down-regulated in the model group/blank group.Conclusion:Shenqi compound which has the function of supplenmenting Qi and Yin and promoting the blood circulation,can inhibit the isletβcell apoptosis,improve isletβcell function,regulate insulin secretion,and prevent T2DM by up-regulating the expression of genes PIK3R1 and Akt1.
作者
刘桠
张翕宇
晁俊
王鹤亭
王钧东
谢春光
LIU Ya;ZHANG Xi-yu;CHAO Jun;WANG He-ting;WANG Jun-dong;XIE Chun-guang(Hospital of Chengdu University of Traditional Chinese Medicine,Chengdu 610072,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2020年第22期34-39,共6页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金项目(81503566)
四川省科技厅重点研发项目(2019YFS0196)
成都中医药大学科技发展基金项目(ZRYY1732)。
