附子、黄连寒热药性对体外足细胞增殖及TRPV4、TRPC6蛋白表达的影响

Effect of Fuzi and Huanglian on in vitro cell proliferation and the expression of TRPC6 and TRPV4

目的研究附子、黄连寒热药性对体外足细胞增殖及TRPV4、TRPC6蛋白表达的影响。方法 MTT法检测足细胞增殖,实验分为:空白对照组、附子组和黄连组。附子组和黄连组加入不同浓度药液进行干预,并测定吸光值(A),计算细胞抑制率。检测TRPV4、TRPC6蛋白试验中,实验分为:空白对照组、附子组和黄连组,其中附子组给予200μg/ml药液、黄连组给予800μg/ml药液,再加入含RPMI 1640培养基培养足细胞24 h。用Western blot法检测附子、黄连对TRPV4、TRPC6蛋白表达影响。结果 MTT的研究表明,黄连组中50,100,200,400,800μg/ml药液均可抑制足细胞生长增殖,且与浓度成正相关。附子在相对较低的质量浓度范围内(<200μg/ml),促进足细胞生长增殖,超过一定实验浓度时(>200μg/ml),抑制足细胞生长增殖。实验表明附子和黄连均可以上调TRPV4蛋白表达,与空白对照组相比差异有统计学意义(P<0.05)。而在TRPC6蛋白表达中,附子和黄连具有明显的下调TRPC6蛋白表达的作用,与空白对照组对比差异有统计学意义(P<0.05)。结论附子、黄连这两种不同药性的中药均可以影响与肾脏病相关的蛋白TRPV4、TRPC6表达。

Objective To explore the effects of Fuzi and Huanglian on podocyte proliferation and the expression of TRPV4 and TRPC6 proteins in vitro. Methods（1）MTT method was used to detect the podocyte proliferation. The experiment was divided into blank control group,Fuzi group and Huanglian group. Different concentrations of Fuzi and Huanglian were used to treat the podocytes. The absorbance value（ A） was measured,and the inhibition rate of cells was calculated.（2）The experiment was divided into blank control group,Fuzi（200 μg/ml） group and Huanglian（800 μg/ml） group. The podocytes were given liquid medicine,and then cultured with RPMI 1640 medium for 24 h to detect the expression of TRPV4 and TRPC6 protein by Western blot. Results MTT results showed that 50,100,200,400 and 800 μg/ml Huanglian inhibited the growth and proliferation of podocyte in a concentration-dependent manner. The relatively low concentration of Fuzi（ 〈 200 μg/ml） promoted the growth and proliferation of podocytes,while the relatively high concentration（ 〉 200 μg/ml） inhibited the growth and proliferation of podocytes. The results showed that the expression of TRPV4 protein in Fuzi group and Huanglian group were significantly increased compared with blank control group（ P 〈 0. 05）,while the expression of TRPC6 protein in Fuzi and Huanglian group were decreased（ P 〈 0. 05）. Conclusion The traditional Chinese medicine Fuzi and Huanglian all have the influence on the expression of TRPV4 and TRPC6 in kidney disease.