绿原酸抑制TGF-β1诱导的人肾小管上皮细胞系HK-2纤维化因子表达

基础医学与临床 ›› 2021, Vol. 41 ›› Issue (12): 1774-1779.

绿原酸抑制TGF-β1诱导的人肾小管上皮细胞系HK-2纤维化因子表达

孙梦奎, 李守林^*

深圳市儿童医院泌尿外科,广东深圳 518038

收稿日期:2021-01-11 修回日期:2021-08-24 发布日期:2021-12-03
通讯作者: *etyylishoulin@163.com
基金资助:
深圳市医疗卫生三名工程(SZSM201612013);深圳市重点专科(2020-2024)(SZXK035)

Chlorgenic acid inhibits TGF-β1-induced fibrotic factors expression of human renal tubular epithelial cell line HK-2

SUN Meng-kui, LI Shou-lin^*

Department of Urology, Shenzhen Children’s Hospital, Shenzhen 518038, China

Received:2021-01-11 Revised:2021-08-24 Published:2021-12-03
Contact: *etyylishoulin@163.com

摘要/Abstract

摘要： 目的探讨绿原酸对TGF-β1诱导的人肾小管上皮HK-2细胞表达纤维化因子的变化及其分子机制。方法体外培养HK-2细胞。用CCK-8法检测细胞增殖;用免疫细胞荧光法检测collagen Ⅰ、collagen Ⅲ的表达;RT-qPCR检测纤维化相关基因mRNA的表达;用Western blot检测TLR4/NF-κB信号通路相关蛋白的表达。结果在不同时间节点(0、12、24、36、72 h)各浓度(0、20、40、80 μg/mL)CGA组细胞增殖无显著差异。与对照组相比,TGF-β1处理组collagen Ⅰ、collagen Ⅲ、α-SMA、vimentin、TGF-β1、snail及slug的表达比对照组明显增加(P＜0.05),但E-cadherin的表达降低(P＜0.05)。CGA能够显著降低TGF-β1对上述分子表达的影响。同时,与TGF-β1处理组相比,CGA能够降低HK-2细胞TLR4、p-IκBα及p-NF-κB蛋白表达(P＜0.05)。结论 CGA能够抑制TGF-β1诱导的人肾小管上皮HK-2细胞表达纤维化因子的增加,其分子机制与调节TLR4/NF-κB信号通路有关。

关键词: 绿原酸, 人肾小管上皮细胞, 纤维化因子, TLR4/NF-κB信号通路

Abstract: Objective To explore the effects of Chlorgenic acid (CGA) on TGF-β1 induced the change of human renal tubular epithelial HK-2 cell fibrotic factors change and the underlying mechanism. Methods HK-2 cells were cultured in vitro. CCK-8 assay was conducted to detect cell proliferation. Immunofluorescence assay was used to detect the expression of collagen Ⅰ and collagen Ⅲ. RT-qPCR was used to detect mRNA expression of HK-2 cell fibrosis related genes. Western blot assay was carried out to detect protein expression of TLR4/NF-κB pathway. Results The proliferation rate of cells in different CGA treated groups at different time points showed no differences. The expression of collagen Ⅰ,collagen Ⅲ,α-SMA,vimentin,TGF-β1,snail and slug increased while E-cadherin decreased in TGF-β1 group(P＜0.05). CGA reversed these expression changes induced by TGF-β1. Further, CGA inhibited TGF-β1 induced TLR4, p-IκBα and p-NF-κB expression (P＜0.05). Conclusions Chlorgenic acid can inhibit TGF-β1 induced increase of fibrotic factors expression in human renal tubular epithelial HK-2 cell through suppressing activation of TLR4/NF-κB pathway.

Key words: chlorgenic acid, human renal tubular epithelial cell, fibrotic factor, TLR4/NF-κB signal pathway

中图分类号:

孙梦奎, 李守林. 绿原酸抑制TGF-β1诱导的人肾小管上皮细胞系HK-2纤维化因子表达[J]. 基础医学与临床, 2021, 41(12): 1774-1779.

SUN Meng-kui, LI Shou-lin. Chlorgenic acid inhibits TGF-β1-induced fibrotic factors expression of human renal tubular epithelial cell line HK-2[J]. Basic & Clinical Medicine, 2021, 41(12): 1774-1779.

参考文献

[1]Wang YC, Dong J, Nie J, et al. Amelioration of bleomycin-induced pulmonary fibrosis by chlorogenic acid through endoplasmic reticulum stress inhibition[J]. Apoptosis, 2017, 22:1147-1156.
[2]Yang F, Luo L, Zhu ZD, et al. Chlorogenic acid inhibits liver fibrosis by blocking the miR-21-Regulated TGF-beta1/Smad7 signaling pathway in vitro and in vivo[J]. Front Pharmacol, 2017, 8:929. doi: 10.3389/fphar.2017.00929.
[3]Liang N, Kitts DD. Role of chlorogenic acids in controlling oxidative and inflammatory stress conditions[J]. Nutrients, 2015, 8:16. doi: 10.3390/nu8010016.
[4]Liu YJ, Zhou CY, Qiu CH, et al. Chlorogenic acid induced apoptosis and inhibition of proliferation in human acute promyelocytic leukemia HL60 cells[J]. Mol Med Rep, 2013, 8:1106-1110.
[5]Shi H, Dong L, Jiang J, et al. Chlorogenic acid reduces liver inflammation and fibrosis through inhibition of toll-like receptor 4 signaling pathway[J]. Toxicology, 2013, 303:107-114.
[6]Sun Z, Ma Y, Chen F, et al. miR-133b and miR-199b knockdown attenuate TGF-beta1-induced epithelial to mesenchymal transition and renal fibrosis by targeting SIRT1 in diabetic nephropathy[J]. Eur J Pharmacol, 2018, 837:96-104.
[7]楚非, 邹万忠, 孙锁柱,等. 大鼠肾小管间质纤维化中肾小管上皮细胞表型转化的研究[J]. 中华肾脏病杂志, 2003, 19:10-14.
[8]Strutz F. Muller GA Renal fibrosis and the origin of the renal fibroblast[J]. Nephrol Dial Transplant, 2006, 21:3368-3370.
[9]Sun YB, Qu X, Caruana G, et al. The origin of renal fibroblasts/myofibroblasts and the signals that trigger fibrosis[J]. Differentiation, 2016, 92:102-107.
[10]Grande MT, Sanchez-Laorden B, Lopez-Blau C, et al. Snail1-induced partial epithelial-to-mesenchymal transition drives renal fibrosis in mice and can be targeted to reverse established disease[J]. Nat Med, 2015, 21:989-997.
[11]方复, 毛华辉, 卢国春, 等. IL-8在胃癌组织上皮间质转化过程中表达增强[J]. 基础医学与临床, 2019, 39:1444-1448.
[12]Sierra-Mondragon E, Molina-Jijon E, Namorado-Tonix C, et al. All-trans retinoic acid ameliorates inflammatory response mediated by TLR4/NF-kappaB during initiation of diabetic nephropathy[J]. J Nutr Biochem, 2018, 60:47-60.
[13]Liu B, Ding F, Hu D, et al. Human umbilical cord mesenchymal stem cell conditioned medium attenuates renal fibrosis by reducing inflammation and epithelial-to-mesenchymal transition via the TLR4/NF-kappaB signaling pathway in vivo and in vitro[J]. Stem Cell Res Ther, 2018, 9:7. doi: 10.1186/s13287-017-0760-6.

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