摘要
目的探究软脂酸(PA)诱导人肾小管上皮细胞(RTEC)炎症和上皮间质转化(EMT)的分子机制。方法用RTEC制备细胞脂质沉积模型,细胞分为空白对照组、牛血清白蛋白(BSA)组、PA组、PA联合干扰素刺激因子(STING)的特异性抑制剂H151组。油红O染色观察RTEC脂质沉积情况,实时定量PCR检测RTEC的白细胞介素6(IL-6)、IL-8、转化生长因子β1(TGF-β1)、1型胶原蛋白α1链(COL1A1)的mRNA水平,Western blot法检测STING、核因子κB p65(NF-κB p65)、磷酸化NF-κB p65(p-NF-κB p65)、TGF-β1、1型胶原蛋白(Col1)的蛋白表达,免疫荧光细胞化学染色检测RETC的Col1表达和分布。结果与对照组相比,PA促进RTEC脂质沉积,显著促进STING表达及NF-κB p65磷酸化,显著上调IL-6、IL-8、TGF-β1、COL1A1的mRNA水平,增加TGF-β1、Col1的蛋白表达和Col1的分布;与PA组相比,H151处理后STING表达及NF-κB p65磷酸化显著降低,IL-6、IL-8、TGF-β1、COL1A1的mRNA水平显著下调,TGF-β1、Col1的蛋白表达和Col1分布减少。结论PA诱导RTEC脂质沉积,激活环鸟腺苷酸合成酶(cGAS)/STING通路,促进其炎症和EMT。
Objective To investigate the molecular mechanism of palmitic acid(PA)inducing inflammation and epithelial to mesenchymal transdifferentiation(EMT)in human renal tubular epithelial cells(RTECs).Methods The cell lipid accumulation model was prepared by RTECs and the cells were divided into blank control group,bovine serum albumin(BSA)group,PA group,and PA combined with stimulator of interferon genes(STING)specific inhibitor H151 group.The lipid accumulation of RTECs were detected by oil red O staining.Real-time quantitative PCR was used to detect the mRNA levels of interleukin 6(IL-6),IL-8,transforming growth factor Bl(TGF-Bl),and type 1 collagen alpha 1 chain(COLIAI)in RTECs.The protein expressions of STING,nuclear factor-kB p65(NF-kB p65),phosphorylated NF-κB p65(p-NF-KB p65),TGF-β1,and type 1 collagen(Coll)were detected by Westem blot and the expression and distribution of Coll in RETCs were detected by immunofluorescence chemical staining.Results Compared with the control group,PA stimulated the lipid deposition,the expression of STING,and the phosphorylation of NF-kB p65 obviously,up-regulated the mRNA levels of IL-6,IL-8,TGF-BI,and COLIAI significantly,increased the protein expressions of TGF-Bl and Coll and the distribution of Coll in RTECs;compared with those in the PA group,after H151 treatment,the expression of STING and the phosphorylation of NF-xB p65 decreased notably,the mRNA levels of IL-6,IL-8,TGF-B1,and COLIA1 were down-regulated dramatically,and the protein expressions of TGF-BI and Coll declined with reduced distribution of Coll.Conclusion PA induces lipid deposition,activated the cyclic guanosine monophosphate-adenosine monophosphate synthase(CGAS)/STING pathway,and caused inflammation and EMT in RTECs.
作者
贺贵贵
王留利
井高静
岑梦佳
赵楠
汤旭磊
HE Guigui;WANG Liuli;JING Gaojing;CEN Mengjia;ZHAO Nan;TANG Xulei(First Clinical Medical College,Lanzhou University,Lanzhou 730000,China;Endocrinology Department of the First Hospital,Lanzhou University,Lanzhou 730000,China)
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2022年第5期385-390,共6页
Chinese Journal of Cellular and Molecular Immunology