益气解毒活络中药对高糖高脂诱导的人肾小球系膜细胞增殖及TNF-α、IL-6表达的作用影响

Effect of Chinese Drugs for Benefiting Qi,Relieving Toxin and Activating Blood on Proliferation and Expressions of TNF-α and IL-6 in Human Glomerular Mesangial Cells Induced by High Glucose and High Lipid

目的观察高糖高脂因素诱导的HBZY-1细胞增殖分化及MMP-9、TNF-α、IL-6基因及蛋白活性表达的变化,探讨益气解毒活络中药防治DN的分子机制。方法体外培养HBZY-1,并根据不同干预因素分为正常组,高糖高脂模型组,第一组,第二组以及第三组(前期实验中效果最佳的两组和理论最佳的一组)共5组。分别以MMT法检测24、48、72 h 3个时间点HBZY-1细胞增殖分化情况;qRT-PCR法检测48 h各组MMP-9、TNF-α及IL-6基因表达;ELISA法检测48 h各组MMP-9、TNF-α及IL-6蛋白活性表达。结果与正常组比较,模型组HBZY-1细胞增殖显著(P<0.01),且在48 h到达峰值,MMP-9基因及蛋白活性表达明显下降(P<0.01)而TNF-α、IL-6表达明显升高(P<0.01);与模型组比较,各用药组能显著抑制细胞增殖(P<0.01),上调MMP-9基因及蛋白活性表达(P<0.01),抑制TNF-α、IL-6表达(P<0.01),以第三组效果最佳(理论最佳组)。结论益气解毒活络中药防治DN的机制与上调MMP-9表达,下调TNF-α、IL-6表达有关。

Objective To observe the proliferation and differentiation of HBZY-1 cells induced by high glucose and high fat factors and the expressions of MMP-9, TNF-α and IL-6 genes and protein activities and explore the molecular mechanism of Chinese drugs for benefiting Qi,relieving toxin and activating blood for preventing and treating DN. Methods HBZY-1 cells were cultured in vitro and divided into normal group, high glucose and high fat model group, the first group, the second group and the third group according to different intervention factors(the best two groups in the previous experiment and the best theoretical theory group). The proliferation and differentiation of HBZY-1 cells were detected by MMT method at 24, 48 and 72 h. The expressions of MMP-9, TNF-α and IL-6 in 48 h of all groups were detected by qRT-PCR. The proteins expressions of MMP-9, TNF-α and IL-6 in 48 h were detected by ELISA. Results Compared with the normal group, the proliferation of HBZY-1 cells in the model group was significant(P<0.01), and reached the peak at 48 h. The expression of MMP-9 gene and protein activity decreased significantly(P<0.01) and the expressions of TNF-α and IL-6 significantly increased(P<0.01). Compared with the model group, each drug group’s inhibited cell proliferation significantly(P<0.01), MMP-9 gene and protein activity were increased(P<0.01) and expressions of TNF-α and IL-6 were inhibited significantly(P<0.01). The third group was the best(the best theoretical group). Conclusion The mechanism of Chinese drugs for benefiting Qi,relieving toxin and activating blood for preventing and treating DN is related to up-regulation of MMP-9 expression and down-regulation of expressions of TNF-α and IL-6.