摘要
目的本研究以罗格列酮为对照,探讨新型PPARγ激动剂CMHX008对高糖诱导人肾小管上皮HK-2细胞纤维化标志的影响及机制。方法罗格列酮和CMHX008处理高糖培养的HK-2细胞,CCK-8法检测不同药物浓度处理后的细胞存活率;免疫印迹法检测纤维化标志蛋白和磷酸化PPARγ(Ser273)的表达;qRT-PCR检测纤维化指标mRNA的表达;划痕和Transwell侵袭实验检测HK-2细胞迁移和侵袭能力的变化。结果CCK-8法结果显示,3μmol·L^-1罗格列酮和3μmol·L^-1CMHX008对HK-2细胞均无明显细胞毒性;免疫印迹法显示,给予罗格列酮和CMHX008能逆转高糖条件下PPARγ(Ser273)磷酸化的上调,减少TGF-β1、α-SMA的表达,增加E-cadherin的表达;划痕和Transwell实验显示,罗格列酮和CMHX008可以抑制高糖引起HK-2细胞迁移和侵袭能力的增强。结论与罗格列酮类似,新型PPARγ激动剂CMHX008能抑制高糖引起的HK-2细胞纤维化标志物的上调,减弱细胞的迁移和侵袭能力,提示CMHX008有改善糖尿病引起肾脏纤维化的作用,其机制可能与抑制PPARγ(Ser273)磷酸化有关。
Aim To investigate the effect of CMHX008,a novel peroxisome proliferator-activated receptor γ(PPARγ) partial agonist,on fibrogenic pathways and its potential mechanism in renal tubular epithelial HK-2 cells in comparison with rosiglitazone.Methods HK-2 cells were cultured with 30 mmol·L^-1 D-(+)-glucose,and then treated with rosiglitazone and CMHX008.Cell counting kit-8(CCK-8) was used to detect cell viability;immunoblotting was used to detect protein expression fibrogenic markers and p-PPARγ(ser273) in HK-2 cells;quantitative real time PCR(qRT-PCR) was used to detect fibrosis-related mRNA levels;wound healing assay and transwell assay were used to detect the migration and invasion ability of HK-2 cells.Results CCK-8 analysis showed that 3 μmol·L ^-1 rosiglitazone and 3 μmol·L ^-1 CMHX008 had no obvious cytotoxicity to HK-2 cells;immunoblotting revealed that rosiglitazone and CMHX008 could reverse the up-regulation of p-PPARγ(ser273),reduce transforming growth factor β1(TGF-β1) and α-smooth muscle actin(α-SMA) protein levels,and up-regulate E-cadherin protein expression levels in HK-2 cells in high glucose conditions;wound healing assay and transwell assay showed that rosiglitazone and CMHX008 could inhibit the increase of migration and invasion ability of hyperglycemia-cultured HK-2 cells.Conclusions The novel PPARγ agonist CMHX008 can improve hyperglycemia-induced renal tubular fibrosis,which may be possibly related to the inhibition of p-PPARγ(Ser273) phosphorylation.
作者
刁俊玲
伍敏
黄荣凤
廖茂霖
陈春秀
李佳渝
李继斌
肖晓秋
DIAO Jun-ling;WU Min;HUANG Rong-feng;LIAO Mao-lin;CHEN Chun-xiu;LI Jia-yu;LI Ji-bin;XIAO Xiao-qiu(Chongqing Key Lab of Translational Medicine in Major Metabolic Disease,the First Affiliated Hospital of Chongqing Medical University,Chongqing Medical University,Chongqing 400016,China;School of Public Health and Management,Chongqing Medical University,Chongqing 400016,China)
出处
《中国药理学通报》
CAS
CSCD
北大核心
2019年第10期1363-1369,共7页
Chinese Pharmacological Bulletin
基金
国家自然科学基金面上项目(No 81570763)
重庆市基础与前沿研究计划项目(No cstc2015 jcy jBX0132)