化合物21(C21)抑制晚期糖基化终产物刺激引起的大鼠肾小管上皮细胞的细胞因子分泌

C21 inhibits cytokine secretion in rat renal tubular epithelial cells stimulated by advanced glycation end products

目的研究2型血管紧张素Ⅱ受体(AT2R)激动剂化合物21(C21)对晚期糖基化终产物(AGE)刺激的NRK-52E大鼠近端肾小管上皮细胞分泌的细胞因子的影响及潜在机制。方法培养NRK-52E细胞,分为正常对照组和(25、50、100、200)mg/L AGE-牛血清白蛋白(BSA)刺激组。培养48 h后收集细胞,采用实时定量PCR检测NRK-52E细胞白细胞介素6(IL-6)、肿瘤坏死因子α(TNF-α)mRNA的表达。间接ELISA检测细胞培养上清液中的IL-6、TNF-α蛋白水平。然后先用25 mg/L AGE-BSA刺激NRK-52E细胞48 h后分别给予(0.01、0.05、0.1)mmol/L C21分别处理24 h,采用实时定量PCR检测细胞蛋白激酶C(PKC)、核因子κB p65(NF-κB p65)及转化生长因子β1(TGF-β1)mRNA的表达,Western blot法检测细胞中PKC、NF-κB p65、TGF-β1蛋白表达。结果不同剂量的AGE-BSA刺激NRK-52E细胞均可明显增加IL-6、TNF-α的mRNA表达,而以25 mg/L AGE-BSA刺激NRK-52E细胞时,其IL-6、TNF-α的蛋白分泌增加更明显。以25 mg/L AGE-BSA刺激NRK-52E细胞48 h后,(0.01、0.05、0.1)mmol/L C21分别处理24 h,均可显著抑制AGE-BSA所诱导的NRK-52E细胞PKC、NF-κB p65及TGF-β1 mRNA和蛋白的表达。结论AGE-BSA促进NRK-52E细胞IL-6、TNF-α、PKC、NF-κB p65及TGF-β1表达,C21则抑制其作用。

Objective To investigate the effect and mechanism of compound 21(C21),an agonist of angiotensin II-2 receptor(AT2R)on the cytokine levels of NRK-52E cells stimulated by advanced glycation end products bovine serum albumin(AGE-BSA).Methods NRK-52E cells were divided into control and(25,50,100,200)mg/L AGE-BSA groups and cultured for 48 hours.The mRNA and protein expression levels of leukin-6(IL-6)and tumor necrosis factorα(TNF-α)were detected by real-time quantitative PCR and ELISA.The NRK-52E cells stimulated by AGE-BSA(25 mg/L)for 48 hours were then treated with(0.01,0.05,0.1)mmol/L C21 for 24 hours.The mRNA and protein expression levels of protein kinase C(PKC),nuclear factor kB p65(NF-kB p65)and transforming growth factorβ1(TGF-β1)were detected by qRT-PCR and Western blot analysis.Results The mRNA expression levels of IL-6 and TNF-αsignificantly increased in NRK-52E cells stimulated by AGE-BSA at diferent doses,with the greatest increase in the 25 mg/L AGE-BSA group.The mRNA and protein expression levels of PKC,NF-kB p65 and TGF-β1 in AGE-BSA-induced NRK-52E cells significantly decreased by(0.01,0.05,0.1)mmol/L C21.Conclusion AGE-BSA promotes the expression of IL-6,TNF-α,PKC,NF-kB p65 and TGF-β1 in NRK-52E cells,while C21 inhibits the effect of AGE-BSA on NRK-52E cells.