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李琰,刘佑晖,蔡虎志,林湘东,陈新宇.温阳振衰颗粒通过调节lncRNA LOC103694972/miR-29c-3p减轻TGF-β1诱导NRK-49F细胞的纤维化[J].湖南中医药大学学报,2023,43(9):1553-1560[点击复制] |
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| 温阳振衰颗粒通过调节lncRNA LOC103694972/miR-29c-3p减轻TGF-β1诱导NRK-49F细胞的纤维化 |
| 李琰,刘佑晖,蔡虎志,林湘东,陈新宇 |
| (湖南中医药大学第一附属医院, 湖南 长沙 410007) |
| 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（Collagen Ⅰ）、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、Collagen Ⅰ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。 |
| 关键词: 温阳振衰颗粒 转化生长因子β1 LOC103694972 miR-29c-3p 纤维化 NRK-49F细胞 |
| DOI：10.3969/j.issn.1674-070X.2023.09.002 |
| 投稿时间：2023-01-03 |
| 基金项目: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）。 |
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| Wenyang Zhenshuai Granules alleviated TGF-β1-induced fibrosis of NRK-49F cells by regulating lncRNA LOC103694972/miR-29c-3p |
| LI Yan,LIU Youhui,CAI Huzhi,LIN Xiangdong,CHEN Xinyu |
| (The First Hospital of Hunan University of Chinese Medicine, Changsha, Hunan 410007, China) |
| Abstract: |
| 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) and α-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 and α-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-49F cell fibrosis. |
| Key words: Wenyang Zhenshuai Granules transforming growth factor β1 LOC10369497 miR-29c-3p fibrosis NRK-49F cell |
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