| 摘要: |
| [目的] 探究加味防己黄芪汤对于致纤维化因子lncRNA NKILA介导的HK-2细胞纤维化模型中上皮细胞-间充质转化（EMT）的干预作用机制。[方法] 基于前期研究基础，使用加味防己黄芪汤高剂量含药血清，干预由lncRNA NKILA过表达慢病毒转染构建的HK-2细胞纤维化模型，并设置AG490干预组进行对照。通过实时荧光定量PCR（qPCR）法、蛋白免疫印迹（Western Blot）法以及免疫荧光法，检测各组lncRNA NKILA的表达量变化、EMT指标FN、Col1、Vim、α-SMA的表达改变以及JAK2/STAT3信号通路指标的变化量。[结果] EMT表型指标检测结果显示，FN、Col1、Vim、α-SMA的mRNA及蛋白的表达量，同过表达阴性对照慢病毒转染组（oe-NC组）比较，过表达lncRNA NKILA慢病毒转染组（oe-NKILA组）表型指标均呈上调趋势，而E-cad的mRNA及蛋白的表达量呈现明显下调趋势；与oe-NKILA组比较，过表达lncRNA NKILA慢病毒转染+AG490干预组（oe-NKILA+AG490组）、过表达lncRNA NKILA慢病毒转染+高剂量防己黄芪汤含药血清组（oe-NKILA+TCM-H）均明显呈下调趋势，而E-cad的mRNA及蛋白的表达量明显上调，差异具有统计学意义（P<0.05）。检测各组lncRNA NKILA的表达量变化结果显示，与oe-NC组比较，oe-NKILA组NKILA表达量显著上调；与oe-NKILA组比较，oe-NKILA+AG490组的NKILA表达量显著下调（P<0.01），oe-NKILA+TCM-H组的NKILA表达量，均明显下调（P<0.05）。JAK2/STAT3通路指标检测结果显示：JAK2、STAT3的mRNA及蛋白、p-JAK2、p-STAT3蛋白表达量，与oe-NC组比较，oe-NKILA组均明显上调，而oe-NKILA+AG490及oe-NKILA+TCM-H组均明显下调，差异具有统计学意义（P<0.05）。[结论] 加味防己黄芪汤高剂量含药血清能够减轻由lncRNA NKILA过表达诱导的HK-2细胞EMT损伤，并且抑制lncRNA NKILA诱导HK-2细胞EMT过程中JAK2/STAT3信号通路的激活。 |
| 关键词: 加味防己黄芪汤 lncRNA NKILA 肾间质纤维化 JAK2/STAT3信号通路 |
| DOI：10.11656/j.issn.1672-1519.2024.12.16 |
| 分类号:R285.5 |
| 基金项目:天津市卫生健康科技项目(TJWJ2024QN071);河北省中医药管理局科研计划项目(T2025046);全国名老中医药专家传承工作室建设项目(978022)。 |
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| To investigate the mechanism by which modified Jiawei Fangji Huangqi Decoction intervenes in the lncRNA NKILA-mediated RIF model of HK-2 cells to inhibit EMT |
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HAN Yu1, WANG Yaogaung2
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1.Tianjin Academy of Traditional Chinese Medicine Affiliated Hospital, Tianjin Institute of Traditional Chinese Medicine, Tianjin 300120, China;2.First Teaching Hospital of Tianjin University of Traditional Chinese Medicine, Tianjin 300381, China
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| Abstract: |
| [Objective] To explore the intervention mechanism of the modified Fangji Huangqi Decoction on the epithelial-mesenchymal transformation in the HK-2 cell fibrosis model mediated by lncRNA NKILA. [Methods] Modified Fangji Huangqi Decoction was applied in an HK-2 cell RIF model built by lncRNA NKILA overexpression lentivirus transfection，with an AG490 control group. qPCR，Western Blot，and immunofluorescence were utilized to assess the alterations in lncRNA NKILA expression，EMT markers FN，Col1，Vim，α-SMA，and JAK2/STAT3 pathway indicators. [Results] EMT phenotypic index analysis indicated up-regulated mRNA and protein levels of FN，Col1，Vim，and α-SMA in the oe-NKILA group versus the oe-NC group，with a noTab.down-regulation in E-cad mRNA and protein levels. Compared to the oe-NKILA group，the oe-NKILA+AG490 and oe-NKILA+TCM-H groups exhibited significant down-regulation in E-cad mRNA and protein levels，with statistical significance（P<0.05）. The expression of lncRNA NKILA in each group was also examined. Compared to the oe-NC group，NKILA expression was significantly elevated in the oe-NKILA group. However，NKILA expression was significantly reduced in the oe-NKILA+AG490 group（P<0.01） and the oe-NKILA+TCM-H group（P<0.05）. The JAK2/STAT3 pathway indicator analysis revealed significant up-regulation in mRNA and protein，p-JAK2，and p-STAT3 protein expressions of JAK2 and STAT3 in the oe-NKILA group versus the oe-NC group，with significant down-regulation in the oe-NKILA+AG490 and oe-NKILA+TCM-H groups（P<0.05）. [Conclusion] The high-dose serum of the modified Fangji Huangqi Decoction can alleviate the EMT injury of HK-2 cells induced by lncRNA NKILA overexpression and inhibit the activation of the JAK2/STAT3 signaling pathway during the process of lncRNA NKILA-inducing EMT of HK-2 cells. |
| Key words: Fangji Huangqi Decoction lncRNA NKILA RIF JAK2/STAT |
