通脉口服液对肾小球系膜细胞增殖及白细胞介素β_1、白细胞介素10表达的影响

Effects of Tongmai Oral Solution on Cell Profliferation and Interleckin-β_1,Interleckin-10 Production in Mesangial Cells

目的：探讨中药复方“通脉口服液”对大鼠肾小球系膜细胞（MsC）增殖及其产生白细胞介素融（IL-β1）和白细胞介素10（IL-10）的影响。方法：应用细胞培养技术进行肾小球系膜细胞的传代培养，采用血清药理学方法，制备通脉口服液含药血清，采用四甲基偶氮唑盐（MTT）法测定通脉口服液含药血清对过度增殖状态下肾小球系膜细胞增殖的影响，酶联免疫吸附法（ELISA）及逆转录聚合酶链反应（RT—PEn）法测定系膜细胞IL—β1 mRNA和IL-10mRNA表达及其蛋白水平。结果：通脉口服液含药血清在5％～20％浓度范围明显抑制脂多糖（LPS）刺激的系膜细胞增殖；在观察的2个时间点（6h、24h）上，LPS均可刺激系膜细胞IL-β1 mRNA的表达及提高IL-β1 分泌水平，在6h时间点上LPS可刺激系膜细胞IL-10mRNA的表达及提高IL-10分泌水平；而通脉口服液在10％浓度上能够明显抑制LPS诱导的系膜细胞IL-β1 分泌及其mRNA的表达，在6h时间点上促进LPS诱导的系膜细胞IL-10分泌及其mRNA的表达。结论：中药复方“通脉口服液”可抑制肾小球系膜细胞的增殖及LPS诱导的系膜细胞IL-β1 的产生，促进LPS诱导的系膜细胞IL-10的产生；肾小球系膜细胞是通脉口服液防治慢性肾炎，延缓肾小球硬化的重要靶细胞之-。

Objective：To explore the effects of the Tongmai Oral Solution on cell proliferation, Interleukin- β1 （IL- β1 ） and Interleukin- 10（ IL- 10） production in mesangial cells. Methods： Cell proliferation was tested by MTT method. The protein synthesis of IL-β1 and IL- 10 in mesangial cells was measured by ELISA. The expression of IL - β1 mRNA and IL - 10 mRNA in mesangial cells was evaluated by RT- PCR. Results： Tongmai Oral Solution （5%-20% ） inhibited the proliferation of mesangial cell. Both mRNA and IL- β1 protein were increased significantly in the presence of LPS and this process could be reversed by Tongmai Oral Solution. Both mRNA and IL - 10 protein were increased significantly in the presence of Tongrnai Oral Solution in 6 hours. Conclusion：The glomerular mesangial cell is one of the main target cell which the Tongmai Oral Solution bring therapeutical action. The possible mechanism of the Tongmai Oral Solution inhibiting the mesangial cell proliferation and deferin4g the renal fibrosis is by decreasing the IL- β1 secretion and gene expression, increasing the IL- 10 secretion and gene expression, blockaging inflammation in mesangial region expansion and circulation furtherly and keeping the dynamic equilibrium between anti - inflammatory cytokine and proinflarnmatory factor.