论文部分内容阅读
目的:研究消癥散结方对高糖刺激下肾小球系膜细胞(MC)的保护作用以及对鞘氨醇激酶1/1磷酸鞘氨醇(Sphk1/S1P)信号通路的干预作用。方法:使用不同浓度消癥散结方含药血清和厄贝沙坦含药血清以及S1PR2抑制剂(JTE-013)与高糖共同培养MC,48h后流式细胞仪检测细胞周期,实时荧光定量PCR和Western Blot法检测FN、ColⅣ、TGF-β1、NF-κB(p65)、Sphk1和S1PR2的表达。结果:与高糖组比较,消癥散结方各剂量组细胞周期中G1期显著增加,S期显著减少(P<0.01);消癥散结方低剂量组细胞FN m RNA和NF-κB(p65)蛋白表达降低(P<0.05),ColⅣ蛋白、Sphk1 m RNA和蛋白表达显著降低(P<0.01);消癥散结方中剂量组细胞Sphk1m RNA表达显著降低(P<0.05),FN m RNA和Sphk1蛋白表达显著降低(P<0.01);消癥散结方高剂量组细胞,FN蛋白、NF-κB(p65)m RNA和蛋白表达显著降低(P<0.05),FN、ColⅣ、Sphk1与S1PR2的m RNA以及TGF-β1的m RNA和蛋白均显著降低(P<0.01)。结论:消癥散结方对高糖刺激下MC的保护作用可能与抑制Sphk1/S1P信号通路有关。
Objective: To investigate the protective effect of Xiaoyasanshuang recipe on glomerular mesangial cells (MCs) stimulated by high glucose and the intervention of Sphk1 / S1P signaling pathway. Methods: The MC-MCs were cultured with different concentrations of Xiao-San San-Fang and irbesartan-containing serum and S1PR2 inhibitor (JTE-013). The cell cycle was detected by flow cytometry after 48 h. Real-time fluorescence quantitative The expression of FN, ColⅣ, TGF-β1, NF-κB (p65), Sphk1 and S1PR2 were detected by PCR and Western Blot. Results: Compared with the high-glucose group, the cell cycle of Xiao-San-San-Fang prescription group increased significantly in G1 phase and significantly decreased in S phase (P <0.01). The FN m RNA and NF-κB (P <0.05). The expression of ColⅣ protein and Sphk1 mRNA and protein were significantly decreased (P <0.01). The expression of Sphk1mRNA in the medium dosage group of Xiaoxusanjieduan group was significantly decreased (P <0.05) (P <0.01). The expression of FN, NF-κB (p65) m RNA and protein were significantly decreased in high dose XHD group (P <0.05) Both m RNA and protein and mRNA of Sphk1 and S1PR2 were significantly decreased (P <0.01). Conclusion: The protective effect of Xiaoxusanjie recipe on MC in high glucose may be related to the inhibition of Sphk1 / S1P signaling pathway.