玄参水提取物对高糖暴露条件下INS-1细胞AMPK的激活作用  

Effect of Aqueous Extract of Scrophularia ningpoensis on AMPK Activity in INS-1 Cells under High Glucose Exposure

作　　者：郭旭 周俊 李晓晗 陈仕琦 高艳果 张永红 王启斌 郑涛[1,2,3] 陈黎 GUO Xu;ZHOU Jun;LI Xiaohan;CHEN Shiqi;GAO Yanguo;ZHANG Yonghong;WANG Qibin;ZHENG Tao;CHEN Li(School of Pharmacy,Hubei University of Medicine,Shiyan 442000,China;Institute of Wudang Traditional Chinese Medicine,Taihe Hospital,Hubei University of Medicine,Shiyan 442000,China;Department of Pharmacy,Taihe Hospital,Hubei University of Medicine,Shiyan 442000,China)

机构地区：[1]湖北医药学院药学院,十堰442000 [2]湖北医药学院附属太和医院武当中医药研究所,十堰442000 [3]湖北医药学院附属太和医院药学部,十堰442000

出　　处：《医药导报》2024年第6期850-854,共5页Herald of Medicine

基　　金：国家自然科学基金青年基金资助项目(81703582);湖北省自然科学基金资助项目(2020CFB713,2016CFB153);湖北省卫生健康科研基金资助项目(WJ2021ZH0017,WJ2019M057);“十四五”湖北省高等学校优势特色学科群(生物与医药)项目(2023BMXKQT3,2022BMXKQT3);湖北医药学院研究生科技创新项目(YC2022056,YC2023068,YC2023069);吴阶平医学基金会临床科研专项基金资助项目(320.6750.2023-25-7)。

摘　　要：目的研究玄参水提取物(AESN)对高糖(HG)条件下INS-1细胞腺苷酸活化蛋白激酶(AMPK)活性的影响。方法将INS-1细胞培养于HG培养基中,并给予不同浓度AESN共孵育处理;利用细胞计数试剂-8(CCK-8)法检测AESN干预对细胞增殖/活力和焦亡小体形成的影响,使用Western blotting法观察AESN对细胞内AMPK表达及磷酸化水平的影响;利用时间分辨-荧光共振能量转移(TR-FRET)实验检测AESN对AMPK激酶活性的影响。结果CCK-8检测结果显示同正常培养条件相比,HG暴露显著降低INS-1细胞增殖/活力并增加焦亡小体形成数量,Western blotting检测结果表明HG暴露可导致细胞内AMPK磷酸化水平下降;而AESN共孵育可呈浓度依赖性地增加INS-1细胞增殖/活力、抑制焦亡小体形成并激活AMPK。TR-FRET实验结果表明,AESN可浓度依赖性增加AMPK激酶活性。结论AESN对HG暴露条件下INS-1细胞AMPK具有激活作用。Objective To investigate the effect of aqueous extract of Scrophularia ningpoensis(AESN)on AMP-activated protein kinase(AMPK)activity in INS-1 cells under high glucose(HG)conditions.Methods INS-1 cells were cultured in HG medium and treated with different concentrations of AESN.The influences of AESN on cell viability and pyroptotic body formation were detected using the CCK-8 assay.Western blotting was used to observe the effects of AESN on AMPK expression and phosphorylation.The effect of AESN on AMPK activity was measured using time-resolved fluorescence resonance energy transfer(TR-FRET)assay.Results Compared with normal glucose controls,INS-1 cells exposed to HG showed decreased cell viabilities and increased formation of pyroptotic bubbles using CCK-8 assay.Western blotting results indicated that HG exposure led to a decrease in intracellular AMPK phosphorylation levels.However,co-incubation with AESN under HG exposure AESN treatment increased the cell viabilities and phosphorylation of AMPK and decreased the pyroptotic bubbles formation in a concentration-dependent manner.In addition,the results of TR-FRET revealed that AESN treatment effectively increased AMPK activity in a concentration-dependent manner.Conclusion AESN has an activating effect on AMPK activity in HG-treated INS-1 cells.

关 键 词：玄参 腺苷酸活化蛋白激酶 高糖暴露 INS-1细胞 荧光共振能量转移 

分 类 号：R285.5[医药卫生—中药学]