摘要
目的探讨芦荟提取物芦荟素(aloin)对TGF-β诱导的肾小管上皮细胞(HK-2)纤维化发生的作用和分子机制。方法实验分为对照组、TGF-β诱导组、芦荟素50μmol·L^(-1)+TGF-β诱导组、芦荟素100μmol·L^(-1)+TGF-β诱导组。通过CCK-8细胞毒性实验检测芦荟素对细胞活力的影响。Western blot、细胞免疫荧光检测纤维化相关蛋白及抗氧化应激相关蛋白表达水平。流式细胞术检测ROS,SOD活性和MDA脂质氧化试剂盒检测氧化应激相关指标。ELISA检测炎症因子水平。结果芦荟素处理HK-2细胞后无显著性影响细胞活性。芦荟素减低TGF-β诱导的HK-2细胞α-SMA、E-Cadherin、Collagen I、Fibronectin纤维化相关蛋白表达水平。芦荟素上调Nrf2、HO-1、NQO1等抗氧化应激分子表达水平,减轻ROS、MDA含量,升高SOD酶活性。同时,芦荟素还能抑制IL-1β、IL-6炎症因子释放。结论芦荟素通过上调Nrf2蛋白表达,改善TGF-β诱导的HK-2细胞氧化应激和纤维化,并抑制其炎症因子释放。
Aim To investigate the effect of aloin,an aloe extract,on fibrosis of renal tubular epithelial cells(HK-2)induced by TGF-βand the underlying molecular mechanism.Methods The experiment included a control group,TGF-βinduced group,TGF-β+Aloin 50 or 100μmol·L^(-1) group.The effect of aloin on cell viability was detected by CCK-8.The expression of fibrosis and antioxidant-related proteins was analyzed by Western blot and immunofluorescence.The ROS was detected by flow cytometry.The related indexes of oxidative stress were detected by SOD activity kit and MDA lipid oxidation kit.The inflammatory factors were detected by ELISA.Results Aloin did not affect the viability of HK-2 cells.Aloin decreased the expression ofα-SMA,E-Cadherin,collagen I and fibronectin in HK-2 cells induced by TGF-β.Aloin up-regulated the expression of antioxidant molecules of Nrf2,HO-1 and NQO1,but reduced the levels of ROS and MDA,and restored the activity of SOD.Meanwhile,aloin also inhibited the release of IL-1βand IL-6 inflammatory factors.Conclusions Aloin can improve the oxidative stress and fibrogenesis of HK-2 cells induced by TGF-βthrough up-regulating Nrf2,and inhibit releasing inflammatory factors.
作者
黄拥军
李艺
王卫黎
HUANG Yong-jun;LI Yi;WANG Wei-li(Dept of Nephrology,Jiangbei Huang Ni Bang Hospital,Chongqing 401147,China;School of Medicine,Chongqing University,Chongqing 400030,China;Nephrology and Urology Center,Southwest Hospital of Army Medical University(Jiangbei branch),Chongqing 400020,China)
出处
《中国药理学通报》
CAS
CSCD
北大核心
2023年第3期549-554,共6页
Chinese Pharmacological Bulletin
基金
国家自然科学基金资助项目(No 81801634)
重庆市科卫联合项目(No 2020FYYX160)。
