熊果酸通过抑制内质网应激减轻高糖和棕榈酸诱导的人主动脉内皮细胞焦亡的研究  

Ursolic acid alleviates high glucose and palmitic acid-induced human aortic endothelial cells pyroptosis via suppression of endoplasmic reticulum stress

作　　者：王宇斐 刘晓玲[1] 吴艳艳 陈敏[1] 胡晓玲[1] 师锐赞[1] 张轩萍[1] WANG Yu-fei;LIU Xiao-ling;WU Yan-yan;CHEN Min;HU Xiao-ling;SHI Rui-zan;ZHANG Xuan-ping(Department of Pharmacology,Shanxi Medical University,Jinzhong 030619,Shanxi Province,China)

机构地区：[1]山西医科大学药理学教研室,山西晋中030619

出　　处：《中国临床药理学杂志》2023年第12期1733-1737,共5页The Chinese Journal of Clinical Pharmacology

基　　金：山西省自然科学研究面上基金资助项目(202203021211226)。

摘　　要：目的探讨熊果酸对高糖高脂诱导人主动脉内皮细胞损伤的影响和机制。方法将人主动脉内皮细胞随机分为对照组、模型组和低、高剂量实验组。模型组用30 mmol·L^(-1)葡萄糖和0.1 mmol·L^(-1)棕榈酸钠刺激内皮细胞24 h以构建高糖高脂损伤模型;低、高剂量实验组分别预孵1和5μmol·L^(-1)熊果酸4 h,随后加入30 mmol·L^(-1)葡萄糖和0.1 mmol·L^(-1)棕榈酸钠处理24 h;对照组给予正常培养。用蛋白质印迹法检测细胞内质网应激及炎症蛋白、活化胱天蛋白酶^(-1)(cleaved caspase-1)和gasdermin D-N(GSDMD-N)的表达水平,用酶联免疫吸附试验法检测细胞培养基中白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)和IL-6的含量。结果低、高剂量实验组、模型组和对照组的葡萄糖调节蛋白78相对表达水平分别为0.78±0.07、0.76±0.08、1.02±0.02和0.67±0.03,活化转录因子6相对表达水平分别为0.80±0.03、0.72±0.04、0.88±0.03和0.69±0.02,磷酸化蛋白激酶样内质网激酶相对表达水平分别为0.66±0.08、0.64±0.07、0.96±0.04和0.51±0.06,磷酸化肌醇依赖酶-1α相对表达水平分别为0.82±0.03、0.67±0.06、0.95±0.01和0.67±0.06,硫氧还蛋白互作蛋白相对表达水平分别为0.85±0.04、0.80±0.03、0.97±0.02和0.68±0.02,核苷酸结合寡聚结构域样受体蛋白3相对表达水平分别为0.84±0.04、0.70±0.05、1.01±0.03和0.66±0.05,cleaved caspase-1蛋白相对表达水平分别为0.09±0.03、0.08±0.02、0.26±0.05和0.17±0.04,GSDMD-N蛋白相对表达水平分别为0.53±0.10、0.54±0.03、0.89±0.07和0.65±0.11,IL-1β含量分别为(3.02±0.24)、(2.51±0.23)、(4.73±0.32)和(2.00±0.16)pg·mL^(-1),TNF-α含量分别为(26.97±2.65)、(22.32±2.50)、(35.18±2.21)和(16.86±1.61)pg·mL^(-1),IL-6含量分别为(91.45±7.12)、(74.22±6.46)、(104.90±4.60)和(49.34±3.94)pg·mL^(-1)。低、高剂量实验组和对照组的上述指标与模型组比较,差异均有统计学意义(P<0.05,P<Objective To investigate the effect and mechanism of ursolic acid(UA)on human aortic endothelial cells(HAECs)injury induced by high glucose and high fat.Methods HAECs were randomly divided into control group,model group,experimental-L group and experimental-H group.The endothelial cells in the model group were stimulated with 30 mmol·L^(-1)glucose and 0.1 mmol·L^(-1)sodium palmitate for 24 h to establish a high glucose/palmitate injury model.The experimental-L group and experimental-H group were pre-incubated with 1,5μmol·L-ursolic acid for 4 h,followed by 30 mmol·L^(-1)glucose and 0.1 mmol·L^(-1)sodium palmitate for 24 h.The control group was given normal culture.The expression levels of endoplasmic reticulum stress and inflammation-related proteins,cleaved caspase-1 and gasdermin D-N(GSDMD-N)were detected by Western blot.The enzyme linked immunosorbent assay used to analyze interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α)and IL-6 in supernatant following to the manufacturer's instructions.Results The relative expression levels of glucose-regulated protein 78 in experimental-L,-H groups,model group and control group were 0.78±0.07,0.76±0.08,1.02±0.02 and 0.67±0.03;the relative expression levels of activating transcription factor 6 were 0.80±0.03,0.72±0.04,0.88±0.03 and 0.69±0.02;the relative expression levels of phospho-protein kinase R-like ER kinase were 0.66±0.08,0.64±0.07,0.96±0.04 and 0.51±0.06;the relative expression levels of phospho-inositol-requiring enzyme 1αwere 0.82±0.03,0.67±0.06,0.95±0.01 and 0.67±0.06;the relative expression levels of thioredoxin interaction protein were 0.85±0.04,0.80±0.03,0.97±0.02 and 0.68±0.02;the relative expression levels of NOD-like receptor protein 3 were 0.84±0.04,0.70±0.05,1.01±0.03 and 0.66±0.05;the relative expression levels of cleaved caspase-1 were 0.09±0.03,0.08±0.02,0.26±0.05 and 0.17±0.04;the relative expression levels of GSDMD-N were 0.53±0.10,0.54±0.03,0.89±0.07and 0.65±0.11;IL-1βcontents were(3.02±0.24),(2.51±0.

关 键 词：熊果酸 内质网应激 焦亡 人主动脉内皮细胞 高糖高脂 炎症 

分 类 号：R28[医药卫生—中药学]