基于PERK-ATF4-CHOP通路探讨马里苷对高糖高脂诱导胰岛β细胞损伤的影响  

Effects of marein on pancreaticβcell injury induced by high glucose and high lipids based on PERK-ATF4-CHOP pathway

作　　者：克迪热亚·卡地尔 祖力皮亚·阿布拉 马红梅[2] 李甜[3] 兰怡[4] 毛新民 Kedireya·Kadier;Zulipiya·Abula;MA Hong-mei;LI Tian;LAN Yi;MAO Xin-min(College of Pharmacy,Xinjiang Medical University,Urumqi 830011;College of Traditional Chinese Medicine,Xinjiang Medical University,Urumqi 830011;College of Basic Medicine,Xinjiang Medical University,Urumqi 830011;Affiliated Traditional Chinese Medicine Hospital of Xinjiang Medical University,Urumqi 830011;Key Laboratory of High Incidence Diseases in Xinjiang,Ministry of Education,Urumqi 830011)

机构地区：[1]新疆医科大学药学院,乌鲁木齐830011 [2]新疆医科大学中医学院,乌鲁木齐830011 [3]新疆医科大学基础医学院,乌鲁木齐830011 [4]新疆医科大学附属中医医院,乌鲁木齐830011 [5]新疆地区高发疾病研究教育部重点实验室,乌鲁木齐830011

出　　处：《中南药学》2022年第6期1316-1321,共6页Central South Pharmacy

基　　金：国家自然科学基金项目(No.U1303233)。

摘　　要：目的探讨马里苷对高糖高脂诱导损伤胰岛β细胞的保护作用及作用机制。方法MIN6细胞先用不同浓度的马里苷预处理,然后高糖高脂诱导MIN6细胞损伤,采用CCK-8法检测马里苷对MIN6细胞活性的影响;采用ELISA法检测细胞外液中胰岛素含量;采用透射电镜(TEM)观察MIN6细胞超微结构;采用特异性钙离子荧光探针(Fluo-4AM)检测内质网钙离子释放;采用Western blot法检测各组细胞激酶R样内质网激酶(PERK)、P-PERK、活化转录因子4(ATF4)及CCAAT增强子结合蛋白同源蛋白(CHOP)、葡萄糖调节蛋白78(GRP78)的表达情况。结果与模型组(HGHF组)比较,马里苷25~100μmol·L^(-1)给药组细胞活力显著升高;高糖刺激胰岛素分泌量水平显著升高;内质网形态和钙离子稳态逐渐恢复正常;P-PERK(马里苷50、100μmol·L^(-1)),ATF4、CHOP、GRP78(马里苷25~100μmol·L^(-1))蛋白表达水平显著降低。结论马里苷能够改善高糖高脂诱导损伤的小鼠胰岛β细胞内质网应激、恢复胰岛素分泌量和内质网钙稳态,其机制可能与内质网应激PERK-ATF4-CHOP通路有关。Objective To determine the protective effect and mechanism of marein on pancreaticβcell injury induced by high glucose and high lipids.Methods MIN6 cells were pretreated with different concentrations of marein and induced by high glucose and high lipid.CCK-8 was used to detect the effect of marein on the activity of MIN6 cells.Insulin secretion was detected by ELISA.The ultrastructure of MIN6 cells were observed by transmission electron microscopy.Endoplasmic reticulum calcium release was detected by specific calcium probe(Fluo-4AM).Western blot was used to detect the expressions of kinase R-like endoplasmic reticulum kinase(PERK),P-PERK,activated transcription factor 4(ATF4),CCAAT enhancer binding protein(CHOP),and glucose regulatory protein 78(GRP78)in all cell groups.Results Compared with the midel group(HGHF),the cell viability of the marein group(25~100 mol·L^(-1))was significantly increased.The insulin secretion(GSIS)level was significantly increased under high glucose stimulation.The protein expression of P-PERK was significantly reduced in 50~100 mol·L^(-1)marein group.The protein expression levels of ATF4,CHOP and GRP78 were significantly reduced in 25~100 mol·L^(-1)marein group.Conclusion Marein can inhibit endoplasmic reticulum stress of pancreaticβcells injury induced by high glucose and high lipids and restore insulin secretion and endoplasmic reticulum calcium homeostasis,which may be related to PERK-ATF4-CHOP pathway.

关 键 词：马里苷 胰岛Β细胞 糖尿病 内质网应激 PERK-ATF4-CHOP 

分 类 号：R285[医药卫生—中药学]