复方积雪草有效组分干预肾小管上皮细胞Toll样受体4表达的实验研究

Experimental study on effects of component from Centella asiatica on compound TLR4 in mice tubular epithelial cells stimulated by tumor necrosis factor α

目的:探讨复方积雪草有效组分-积雪草苷/大黄素干预肿瘤坏死因子α(TNF-α)诱导的小鼠肾小管上皮细胞(mTEC)Toll样受体4(TLR4)mRNA及蛋白的表达水平。方法:采用mTEC,模型组用TNF-α5ng/mL诱导,治疗组在TNF-α诱导的同时,以不同浓度的积雪草苷合大黄素进行干预,于24h后分别提取细胞RNA及上清,应用逆转录-聚合酶链反应(RT-PCR)和流式细胞术(FCM)分别检测mTECTLR4 mRNA和膜蛋白的表达。结果:正常mTEC具有TLR4 mRNA和蛋白表达,经TNF-α诱导后TLR4表达明显上调,当积雪草苷合大黄素干预浓度达4μg/mL+0.4μg/mL时,能够抑制TLR4 mRNA及蛋白的表达水平。结论:复方积雪草有效组分能够抑制TNF-α上调所致的肾小管上皮细胞TLR4过度表达,从而缓解肾脏局部失控性炎症反应。

AIM： To investigate the effects of component from Centella asiatica （centella asiatica ＆ emodin） on toll like receptor 4 （TLR4） mRNA and protein in mice tubular epithelial cells （mTEC） stimulated by tumor necrosis factor α （TNF-α）. METHODS： Cultured mTEC were exposed to TNF-α 5 ng/mL and centella asiatica ＆ emodin for 24 hours, followed by transcriptase-polymerase chain reaction （RT-PCR） and enzyme-linked flow cytometry （FCM） analysis for complement TLR4 mRNA and protein expression. RESULTS： TLR4 mRNA and protein production were present in mTEC. TNF-α up-regulated TLR4 production in mTEC. Centella asiatica combined with emodin synergistically down-regulate TLR4 expression at a dose of 4ug/mL and 0.4 ug/mL. CONCLUSION： Effective component from Centella asiatica component induced TLR4 stimulated by TNF-α in source of kidney local.