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目的:观察四妙散含药血清对尿酸(UA)诱导的人肾小管上皮细胞(HK-2)TGF-β1、α-SMA表达的影响,探讨其在防治肾间质纤维化方面的作用机制。方法:将HK-2细胞分为空白组,模型组,空白血清组,高、中、低剂量组。药物干预24h后,MTT检测细胞活性抑制率;Real-time PCR检测TGF-β1、α-SMA m RNA的表达;免疫组织化学检测TGF-β1、α-SMA蛋白的表达。结果:MTT检测显示尿酸抑制细胞活性(P<0.01),经含药血清干预后细胞活性抑制明显改善(P<0.01);Real-time PCR与细胞免疫组化检测显示,尿酸刺激后TGF-β1、α-SMA的m RNA和蛋白表达量均明显增加(P<0.05),经含药血清干预后,TGF-β1、α-SMA的表达随着剂量增加逐步下降(P<0.05),呈现一定剂量依赖性。结论:下调TGF-β1、α-SMA的表达,抑制上皮细胞转分化,可能是四妙散抗肾间质纤维化的机制之一。
OBJECTIVE: To observe the effect of Sijiaosan medicated serum on uric acid (UA) -induced TGF-β1 and α-SMA expression in human renal tubular epithelial cells (HK-2) and to explore its mechanism of action in preventing and treating renal interstitial fibrosis . Methods: HK-2 cells were divided into blank group, model group, blank serum group, high, medium and low dose groups. The cytotoxicity was detected by MTT assay after 24h of drug intervention. The expression of TGF-β1 and α-SMA m RNA was detected by Real-time PCR. The expression of TGF-β1 and α-SMA protein was detected by immunohistochemistry. Results: MTT assay showed that uric acid inhibited the cell viability (P <0.01) and inhibited the cell viability significantly (P <0.01) after intervention with serum. The results of Real-time PCR and cell immunohistochemistry showed that uric acid stimulated TGF- , The mRNA and protein expression of α-SMA increased significantly (P <0.05). The expression of TGF-β1 and α-SMA decreased gradually with the increase of dose (P <0.05) Dose-dependent. Conclusion: Down-regulating the expression of TGF-β1 and α-SMA and inhibiting epithelial cell transdifferentiation may be one of the mechanisms of Si Miao Powder's anti-renal interstitial fibrosis.