尿石素A抗糖脂毒性改善MIN6胰岛β细胞活力机制靶点研究  

Study on the mechanism targets of urolithin A anti-glycolipid toxicity improving MIN6 pancreatic β-cell viability

作　　者：张之 董怀洋 古丽米拉•艾斯克尔 ZHANG Yan-zhi;DONG Huai-yang;GULIMILA Aisikeer(Department of Pharmacology,School of Pharmacy,Xinjiang Medical University,Urumqi 830011,Xinjiang Uyghur Autonomous Region,China)

机构地区：[1]新疆医科大学药学院,新疆乌鲁木齐830011

出　　处：《中国临床药理学杂志》2022年第5期395-398,403,共5页The Chinese Journal of Clinical Pharmacology

基　　金：国家自然科学基金资助项目(81760767);新疆维吾尔自治区自然科学基金资助项目(2017D01C204);新疆自治区重点实验室建设基金资助项目(XJDX1713);新疆医科大学博士启动基金资助项目(2019-017)。

摘　　要：目的靶点通路预测及细胞实验验证尿石素A(UA)在糖尿病环境下保护胰岛β细胞的作用机制。方法用Cytoscape软件分析成分药效靶点,以基因本体(GO)、京都基因和基因组百科全书(KEGG)富集分析信号通路。将MIN6胰岛β细胞分为3组:对照组、高糖脂模型组、给药组(UA,11.5μg·mL^(-1)),干预24 h。CCK-8法测定细胞活力(光密度值),蛋白质印迹法检测蛋白激酶B(Akt)、哺乳动物雷帕霉素蛋白(mTOR)及其相应磷酸化蛋白(p-Akt、p-mTOR)表达水平。自噬抑制剂氯喹(CQ,50μmol·L^(-1))、AMP依赖的蛋白激酶抑制剂(CC,10μmol·L^(-1))进行进一步反证。结果网络预测发现Akt、mTOR为主要靶点,KEGG分析得到34条信号通路(P<0.05),Akt-mTOR是其中一条重要通路。对照组、高糖脂模型组、给药组的细胞活力分别为99.92±1.84, 47.40±2.78和67.07±2.95;p-Akt表达分别为1.00±0.02, 0.61±0.01和0.79±0.01;p-mTOR表达分别为1.00±0.08,0.54±0.02和0.83±0.03。UA联合CQ干预后,p-Akt、p-mTOR表达分别为0.69±0.02和0.66±0.08。UA联合CC干预后,p-Akt、p-mTOR表达分别为0.67±0.03和0.80±0.07。模型组与对照组比较,或给药组与模型组比较,UA联合抑制剂给药组与单独UA给药组比较,差异均有统计学意义(均P<0.05)。结论 UA可能通过激活AMPK和自噬靶向Akt/mTOR信号通路改善胰岛β细胞自噬,对抗糖脂毒性损伤,提高胰岛β细胞活力。Objective To verify the mechanism of urolithin A (UA) in protecting pancreaticβcells from diabetic environment.Methods Cytoscape software was used to analyzes the components and drug targets,gene ontology (GO),Kyoto Encyclopedia of Genes and Genomes(KEGG) enriches and analyzes signal pathways.MIN6 pancreatic isletβcells were divided into three groups:control group,high-glycolipid model group,and administration group (UA,11.5μg·m L^(-1))) for 24h.The cell viability(OD value) was determined by CCK-8 method.The expression levels of protein kinase B (Akt),mammalian rapamycin(m TOR) and their corresponding phosphorylated proteins (p-Akt,pm TOR) were detected by Western blotting.Autophagy inhibitor chloroquine (CQ,50μmol·L^(-1))) and AMPK inhibitor (CC,10μmol·L^(-1))) were used for further counter-confirmation.Results Akt,m TOR were main targets analyzed by Cytoscape,34 signal pathway were found by KEGG analysis,Akt-m TOR was an important one among these.The cell viability in control group,high-glycolipid model group,and administration group were 99.92±1.84,47.40±2.78,67.07±2.95;p-Akt level in the three groups were 1.00±0.02,0.61±0.01,0.79±0.01;p-m TOR expression in the three groups were 1.00±0.08,0.54±0.02,0.83±0.03.Co-treatment CQ with UA,p-Akt,p-m TOR level were changed to (0.69±0.02,0.66±0.08);Co-treatment CC with UA,p-Akt,p-m TOR level were changed to (0.67±0.01,0.80±0.09).Comparison between model group and control group,or UA group and model group,or inhibitor combination group and UA alone group,the differences of the factors were significant (all P<0.05).Conclusion The machanism of UA improve viability of pancreaticβ-cell agaist glycolipotoxic by activating AMPK and autophagy targeting the Akt/m TOR signaling pathway.

关 键 词：尿石素A 网络药理学 MIN6胰岛β细胞 蛋白激酶B/哺乳动物雷帕霉素蛋白信号通路 

分 类 号：R28[医药卫生—中药学]