大蒜素通过JAK2/STAT3信号通路改善高糖诱导的人腹膜间皮细胞-间充质转化  

Allicin improves human peritoneal mesenchymal cell-mesenchymal transformation induced by high glucose through the JAK2/STAT3 signaling pathway

作　　者：甘林望 李乾程 高利超 刘琦 李莹 王玉洁 欧三桃 GAN Linwang;LI Qiancheng;GAO Lichao;LIU Qi;LI Ying;WANG Yujie;OU Santao(Department of Nephrology,Affiliated Hospital of Southwest Medical University/Sichuan Clinical Research Center for Nephrology,Luzhou 646000;China.2.Department of Respiratory Medicine,Affiliated Hospital of Southwest Medical University,Luzhou 646000)

机构地区：[1]西南医科大学附属医院肾病内科/四川省肾脏疾病临床医学研究中心,四川泸州646000 [2]西南医科大学附属医院呼吸内科,四川泸州646000

出　　处：《中国比较医学杂志》2023年第11期39-47,共9页Chinese Journal of Comparative Medicine

基　　金：四川省肾脏疾病临床医学研究中心2020年度开放课题(2019YFS0537-16)。

摘　　要：目的探讨大蒜素改善高糖诱导的人腹膜间皮细胞-间充质转化的相关机制。方法培养人腹膜间皮细胞(human peritoneal mesothelial cells,HPMCs)后将其进行两次分组,分组1:①对照组;②8.5 mmol/L D-葡萄糖诱导组(8.5 mmol/L DG组);③17 mmol/L D-葡萄糖诱导组(17 mmol/L DG组);④34 mmol/L D-葡萄糖诱导组(34 mmol/L DG组);⑤68 mmol/L D-葡萄糖诱导组(68 mmol/L DG组)。其中除对照组外,其余组分别用8.5、17、34、68 mmol/L的D-葡萄糖诱导48 h。分组2:①对照组;②34 mmol/L D-葡萄糖诱导组(HG组);③34 mmol/L D-葡萄糖+低剂量大蒜素诱导组(AL-L组);④34 mmol/L D-葡萄糖+中剂量大蒜素诱导组(AL-M组);⑤34 mmol/L-葡萄糖+高剂量大蒜素诱导组(AL-H组);⑥34 mmol/L D-葡萄糖+JAK2抑制剂诱导组(JAK2组)。其中HG组用34 mmol/L的D-葡萄糖诱导48 h,AL-L组、AL-M组、AL-H组用34 mmol/L的D-葡萄糖预处理6 h后分别用10、20和40 ng/mL大蒜素诱导48 h,JAK2组加入1μmol/L AG490预处理6 h后用34 mmol/L的D-葡萄糖诱导48 h。ELISA检测HPMCs上清的IL-6、TNF-α和IL-1β的含量;CCK-8检测细胞增殖并观察形态;Western blot检测JAK2、p-JAK2、STAT3、p-STAT3、N-cadherin、E-cadherin、Vimentin、α-SMA、MCP-1、p65、p-p65蛋白的表达情况。结果与对照组相比,高糖诱导组HPMCs的相对存活率明显降低(P<0.01),细胞形态表现异常,促进上皮细胞-间充质转分化(epithelial-mesenchymal transition,EMT)发生的α-SMA、N-cadherin和Vimentin表达明显上调,抑制EMT发生的E-cadherin蛋白表达明显下调,JAK2/STAT3信号通路被激活从而导致EMT的发生(P<0.01);而大蒜素能明显促进高糖诱导后的HPMCs增殖,恢复异常的细胞形态,调节与EMT发生的相关蛋白水平从而改善HPMCs的上皮间质转分化;与高糖诱导组相比,大蒜素处理组HPMCs的促炎症因子IL-1β、IL-6和TNF-α明显降低,促炎症蛋白p-p50和MCP1表达明显下调,表明大蒜素能改善EMT引起的炎症。结论大蒜素可通�ObjectiveTo investigate the mechanism of allicin in improving human peritoneal mesenchymal cell-mesenchymal transformation induced by high glucose.MethodsHuman peritoneal mesothelial cells(HPMCs)were divided into the following groups.Group 1:①Control group;②8.5 mmol/L D-glucose group(8.5 mmol/L DG group);③17 mmol/L D-glucose group(17 mmol/L DG group);④34 mmol/L D-glucose group(34 mmol/L DG group);⑤68 mmol/L D-glucose group(68 mmol/L DG group).Except in the control group,the groups were treated with the corresponding concentrations of D-glucose for 48 h.Group 2:①Control group;②34 mmol/L D-glucose group(HG group);③34 mmol/L D-glucose+low dose allicin group(AL-L group);④34 mmol/L D-glucose+medium dose allicin group(AL-M group);⑤34 mmol/L glucose+high-dose allicin group(AL-H group);⑥34 mmol/L D-glucose+JAK2 inhibitor group(JAK2 group).The HG group was treated with 34 mmol/L D-glucose for 48 h.AL-L,AL-M and AL-H groups were pretreated with 34 mmol/L D-glucose for 6 h and then treated with 10,20 and 40 ng/mL allicin for 48 h,respectively.The JAK2 group was pretreated with 1μmol/L AG490 for 6 h and then treated with 34 mmol/L D-glucose for 48 h.IL-6,TNF-α,and IL-1βcontents in HPMC culture supernatants were determined by ELISA.A CCK-8 assay was used to assess cell proliferation and morphology.JAK2,p-JAK2,STAT3,p-STAT3,N-cadherin,E-cadherin,Vimentin,α-SMA,MCP-1,p65 and p-p65 protein expression was detected by Western blot.ResultsCompared with the control group,the relative survival rate of HPMCs in the high glucose induced group was significantly reduced(P<0.01),cell morphology was abnormal,expression ofα-SMA,N-cadherin and Vimentin that promote epithelial-mesenchymal transition was significantly upregulated,and expression of E-cadherin,which inhibits EMT,was significantly downregulated.The JAK2/STAT3 signaling pathway was activated,leading to EMT(P<0.01).Allicin significantly promoted HPMC proliferation induced by high glucose,reversed the abnormal cell morphology,regulated the expression of EM

关 键 词：人腹膜间皮细胞 EMT 大蒜素 JAK2/STAT3信号通路 

分 类 号：R-33[医药卫生]