摘要
目的探讨尿酸盐刺激下HK-2肾小管上皮细胞过氧化物酶体增殖物激活型受体-γ(PPAR-γ)表达及替米沙坦的干预作用。方法培养HK-2肾小管上皮细胞,随机分为7组,分别为对照组(NC)、模型组(UM)、替米沙坦组(UT)、罗格列酮组(UR)、GW9662组(UG)、替米沙坦+GW9662组(UTG)和罗格列酮+GW9662组(URG),除外对照组应用培养液外,其余6组在对照组基础上均加入600μmol/L尿酸盐溶液培养。各组刺激培养48 h后,细胞融合约90%~95%。流式细胞仪检测HK-2细胞hUAT、PPAR-γ蛋白表达及细胞凋亡率;Q-PCR及western blot检测HK-2细胞hUAT、PPAR-γ、TGF-β1蛋白表达。结果(1)与NC组相比,UM组肾小管上皮细胞凋亡增加(P<0.05),替米沙坦能减轻凋亡程度(P<0.05),应用PPAR-γ抑制剂后,替米沙坦无法逆转尿酸盐刺激所致HK-2细胞增加(P<0.05)。(2)NC组存在hUAT、PPAR-γ表达,与NC组相比,UM、UG组hUAT、PPAR-γ蛋白阳性细胞率、基因及蛋白表达减少(P<0.05),TGF-β1基因及蛋白表达增加(P<0.05);与UG组相比,UT组、UR组hUAT、PPAR-γ蛋白阳性细胞率、基因及蛋白表达增加(P<0.05),TGF-β1基因及蛋白表达降低(P<0.05),与UT组、UR组分别比较,URG组、UTG组hUAT、PPAR-γ蛋白阳性细胞率、基因及蛋白表达减少(P<0.05),TGF-β1基因及蛋白表达增加(P<0.05)。结论(1)尿酸盐刺激HK-2肾小管上皮细胞PPAR-γ蛋白表达,替米沙坦具有激活PPAR-γ受体作用;(2)替米沙坦能调节尿酸盐刺激肾小管上皮细胞后所致hUAT蛋白下降,起到肾脏保护作用,这种作用是通过PPAR-γ调节肾小管上皮细胞凋亡及TGF-β1蛋白表达实现的。
Objective To investigate PPAR--y expression of urate-stimulated HK-2 renal tubular epithelial cells and the interventional mechanism of telmisartan.Methods HK-2 renal tubular epithelial cells were cultured and randomly divided into 7 groups,which were respectively the control group(NC group),model group(UM),telmisartan group(UT group),rosiglitazone group(UR group),GW9662 group(UG group),telmisartan+GW9662 group(UTG group),and rosiglitazone+GW9662 group(URG group).Except for the control group,which used the culture solution,the remaining 6 groups were added with 600μmol/L urate solution on the basis of the control group.After 48 hours of stimulation culture in each group,the cell fusion was about 90%~95%.Flow cytometry were used to detect hUAT and PPAR-γprotein expression and apoptosis rate in HK-2 cells.Q-PCR and western blot were used to detect hUAT,PPAR-γand TGF-β1 protein expression in HK-2 cells.Result(1)Compared with the NC group,apoptosis of renal tubular epithelial cells increased in the UM group(P<0.05),and telmisartan could reduce the degree of apoptosis(P<0.05).After applying PPAR-γinitiation inhibitor,telmisartan could not reverse the the increased apoptosis of hk-2 cells induced by urate stimulation(P<0.05).(2)The results of several methods consistently showed that hUAT and PPAR-γexpression existed in the NC group;Compared with NC group,the positive cell rate,gene and protein expression of hUAT and PPAR-γin UM group and UG group decreased(P<0.05),while the gene and protein expression of TGF-β1 increased(P<0.05);Compared with UG group,the positive cell rate,gene and protein expression of hUAT and PPAR-γin UT group and UR group increased(P<0.05),TGF-β1 gene and protein expression decreased(P<0.05);Compared with UT group and UR group,the positive cell rate,gene and protein expression of hUAT and PPAR-γin URG group and UTG group decreased(P<0.05),while the gene and protein expression of TGF-β1 increased(P<0.05).Conclusion(1)After urate stimulation of hk-2 renal tubular epithelial cells,hk-2 ce
作者
龙利
张继波
余建峰
覃慧群
金晟
徐敏
LONG Li;ZHANG Jibo;YU Jianfeng;QIN Huiqun;JIN Sheng;XU Min(Department of Nephrology,the Third People′s Hospital of Hubei Province,Wuhan 430033,China)
出处
《实用医学杂志》
CAS
北大核心
2020年第16期2205-2211,共7页
The Journal of Practical Medicine
基金
湖北省自然科学基金(编号:2015CFB676)。
