摘要
目的探讨温阳振衰颗粒通过lncRNA LOC103694972/miR-29c-3p对TGF-β1诱导的大鼠肾间质成纤维细胞(NRK-49F)的纤维化的影响。方法采用TGF-β1(10 ng·mL^(-1))处理NRK-49F细胞24 h,构建纤维化细胞模型。造模结束后给予空白血清、温阳振衰颗粒含药血清和缬沙坦胶囊含药血清干预。CCK-8法检测温阳振衰颗粒对NRK-49F增殖的影响。实时荧光定量PCR检测LOC103694972和miR-29c-3p表达。Western blot法检测纤维化相关因子Ⅰ型胶原(CollagenⅠ)、α-平滑肌肌动蛋白(α-SMA)表达。同时研究过表达或沉默LOC103694972,温阳振衰颗粒含药血清对TGF-β1干预的NRK-49F细胞的影响。双荧光素酶报告实验检测LOC103694972与miR-29c-3p之间的调控作用。结果温阳振衰颗粒明显减弱TGF-β1诱导的细胞增殖和LOC103694972的表达(P<0.05),促进miR-29c-3p的表达(P<0.05),其干预效果与缬沙坦胶囊含药血清效果一致。双荧光素酶实验证实LOC103694972靶向调控miR-29c-3p。过表达LOC103694972促进温阳振衰颗粒含药血清干预下TGF-β1诱导NRK-49F细胞的活力,以及α-SMA蛋白、CollagenⅠ蛋白的表达(P<0.05)。而沉默LOC103694972具有相反的效果(P<0.05)。结论温阳振衰颗粒通过下调LOC103694972促进miR-29c-3p的表达,进而抑制TGF-β1诱导的NRK-49F细胞纤维化。
Objective To explore the effects of Wenyang Zhenshuai Granules on the fibrosis of transforming growth factor-β1(TGF-β1)-induced rat renal interstitial fibroblasts(NRK-49F)through lncRNA LOC103694972/miR-29c-3p.Methods NRK-49F cells were cultured and induced by TGF-β1(10 ng·mL^(-1),24 h)to construct a renal fibrosis cell model.After the modeling,cells were given blank serum,Wenyang Zhenshuai Granules-containing serum and valsartan capsules-containing serum as interventions.CCK-8 assay was used to check the effects of Wenyang Zhenshuai Granules on the proliferation of NRK-49F cells.Real-time fluorescence quantitative PCR was used to check the expressions of LOC103694972 and miR-29c-3p.Western blot was used to examine the expression of fibrosis-related factors Type I collagen(Collagen I)andα-smooth muscle actin(α-SMA).The effects of Wenyang Zhenshuai Granules-containing serum on NRK-49F cells with overexpression or silencing of TGF or LOC103694972 was also studied.Dual luciferase reporting assay was used to test the regulation of LOC103694972 on miR-29c-3p.Results Wenyang Zhenshuai Granules significantly reduced cell proliferation and LOC103694972 expression induced by TGF-β1(P<0.05),and promoted miR-29c-3p expression in TGF-β1-induced NRK-49F cell(P<0.05).The intervention effect was consistent with that of valsartan capsule-containing serum.Dual luciferase reporting assay confirmed the targeted regulation of LOC103694972 on miR-29c-3p.Overexpression of LOC103694972 promoted the vitality of NRK-49F cells induced by TGF-β1 and the expression of Collagen I andα-SMA proteins under the intervention of Wenyang Zhenshuai Granules-containing serum(P<0.05).While silencing LOC103694972 had the opposite effect(P<0.05).Conclusion Wenyang Zhenshuai Granules can promote the expression of miR-29c-3p by down-regulating LOC103694972,thereby inhibiting TGF-β1-induced NRK-49 cell fibrosis.
作者
李琰
刘佑晖
蔡虎志
林湘东
陈新宇
LI Yan;LIU Youhui;CAI Huzhi;LIN Xiangdong;CHEN Xinyu(The First Hospital of Hunan University of Chinese Medicine,Changsha,Hunan 410007,China)
出处
《湖南中医药大学学报》
CAS
2023年第9期1553-1560,共8页
Journal of Hunan University of Chinese Medicine
基金
湖南省教育厅一般项目(20C1412)
湖南省研究生科研创新项目(CX2018B474)
全国名中医药专家陈新宇寒舍传承工作室建设项目(国家中医药人教函〔2022〕75号)。
