论文部分内容阅读
目的探讨山奈酚对高糖诱导大鼠肾系膜细胞(HBZY-1)增殖、纤维化的影响,并研究其相关的作用机制。方法将HBZY-1细胞分为正常糖浓度组[葡萄糖(Glu)5.5mmol/L]即对照组、高糖(Glu 25mmol/L)组、10μmol/L山奈酚+高糖组和30μmol/L山奈酚+高糖组。分组处理细胞后,采用MTT实验检测正常糖浓度和高糖下山奈酚对HBZY-1细胞增殖的影响;流式细胞仪检测各组细胞周期变化;Real-time PCR检测各组细胞纤维连接蛋白(FN)、结缔组织生长因子(CTGF)mRNA的表达,Western blot检测FN、CTGF、细胞周期相关蛋白以及p38 MAPK信号通路蛋白的表达。结果山奈酚(10、30μmol/L)对正常糖浓度培养的HBZY-1细胞增殖影响不大(P>0.05),高糖能够增强细胞的增殖能力(P<0.05),而山奈酚(10、30μmol/L)可抑制高糖引起的增强细胞增殖作用(P<0.05)。高糖能减少G0/G1期细胞比例,增加S期细胞比例,并降低细胞周期相关蛋白p21Cip1以及p27Kip1蛋白表达,山奈酚(10、30μmol/L)能够阻止高糖引起的细胞周期改变以及细胞周期相关蛋白表达的下调。山奈酚(10、30μmol/L)同时能够抑制高糖引起的细胞FN、CTGF mRNA和蛋白表达水平的上调。高糖能够增加磷酸化p38丝裂原活化蛋白激酶(p-p38 MAPK)蛋白水平,而山奈酚(10、30μmol/L)处理能够剂量依赖性的抑制高糖诱导的p-p38MAPK蛋白的表达上调。结论山奈酚对高糖诱导的HBZY-1细胞增殖起抑制作用,其作用机理可能是通过调控p38 MAPK信号通路,对高糖培养下的HBZY-1细胞起到保护作用。
Objective To investigate the effects of kaempferol on proliferation and fibrosis of rat mesangial cells (HBZY-1) induced by high glucose, and to study its mechanism of action. Methods HBZY-1 cells were divided into normal glucose concentration group (Glu, 5.5mmol / L), control group, Glu 25mmol / L group, 10μmol / L kaempferol and high glucose group and 30μmol / Phenol + high glucose group. After grouping the cells, MTT assay was used to detect the effect of kaempferol on the proliferation of HBZY-1 cells under high glucose; the changes of cell cycle were detected by flow cytometry; the expression of fibronectin FN), connective tissue growth factor (CTGF) mRNA expression, Western blot detection of FN, CTGF, cell cycle related protein and p38 MAPK signal pathway protein expression. Results Kaempferol (10,30 μmol / L) had no significant effect on the proliferation of HBZY-1 cells cultured in normal glucose concentration (P> 0.05), and high glucose increased the proliferation of cells (P <0.05) 30μmol / L) inhibited the proliferation of cells induced by high glucose (P <0.05). High glucose could reduce the proportion of cells in G0 / G1 phase, increase the proportion of cells in S phase and decrease the expression of cell cycle related protein p21Cip1 and p27Kip1, and kaempferol (10,30μmol / L) could prevent cell cycle changes induced by high glucose and cell cycle Down-regulation of related protein expression. Kaempferol (10, 30μmol / L) at the same time can inhibit the high glucose induced FN, CTGF mRNA and protein expression levels. High glucose increased the protein level of phospho-p38 mitogen-activated protein kinase (p-p38 MAPK), whereas kaempferol treatment inhibited the high glucose-induced increase of p-p38MAPK protein in a dose-dependent manner . Conclusions Kaempferol can inhibit the proliferation of HBZY-1 cells induced by high glucose. The mechanism may be that it protects HBZY-1 cells under high glucose by regulating p38 MAPK signaling pathway.