摘要
目的探讨甘草甜素(GL)对HepG2.2.15细胞上清HBeAg分泌,Toll样受体4(TLR4)信号分子的表达及对细胞增殖的影响。方法实时荧光定量PCR检测TLR4表达;直接免疫荧光流式细胞术(FCM)检测表达TLR4的阳性细胞率;ELASA检测HBV所分泌抗原;MTT检测细胞增殖活性,并与空白对照组比较。结果给药后HBVe抗原(HBeAg)的分泌结果显示,总体均数间差异显著(P<0.05),但甘草甜素各组与对照组比较差异无统计学意义;GL各组TLR4 mRNA及流式细胞TLR4的表达总体均数间均有显著差异(P<0.01),分别与对照组相比差异有显著性(P<0.01),100μg/mL组尤其明显;MTT实验显示,200μg/mL以下三个剂量组均可促进细胞增殖,50μg/mL组与对照组间差异显著(P<0.05);400μg/mL、800μg/mL两组均显著抑制细胞增殖(P<0.01);而MTT结果与HBeAg水平呈显著负相关(P<0.01),与TLR4表达无相关关系(P>0.05)。结论研究表明,甘草甜素对HepG2.2.15的e抗原分泌可能具有双向作用;细胞的存活率与e抗原呈负相关;TLR4在HepG2.2.15细胞株低表达,GL可上调TLR4表达,甘草甜素可影响先天免疫中的TLR4信号分子,有可能在体内通过免疫途径影响HBV复制和抗原分泌。
【Objective】 To investigate the effect of glycyrrhizin （GL） on the expression of hepatitis B virus e antigen （HBeAg ）, Toll-like receptors 4 （TLR 4） level and cell proliferation in HepG 2.2.15 cell line.【Methods】 TLR4 mRNA level was examined by real-time PCR, the positive percent of cells expressing TLR 4 was examined by FCM before and after stimulation with GL. HBeAg level was examined by ELASA and cell proliferation was examined by MTT. The GL groups were compared with the control group.【Results】 HBeAg levels after stimulation with GL were significantly difference from that before stimulation （P 0.05）. HBeAg level in the 800 μg/mL group increased significantly compared with those in the 100 and 200μg/mL groups （P 0.05）. The intensity of TLR4- positive cells and TLR4 mRNA levels in the GL groups were significantly difference from those in the control group （P 0.05）, that in the 100 μg/mL group increased significantly compared with those in other groups. GL equal to or below 200 μg/mL could promote cell proliferation, but only that in the 50 μg/mL group significantly increased cell proliferation compared with the control group （P 0.05）. In the 400 μg/mL and 800 μg/mL groups, the growth of cells was inhibited significantly （P 0.01）, cell proliferation was notably negatively correlated with the expression of HBeAg （P 0.05）, but was not correlated with TLR4 levels.【Conclusion】 GL could inhibit or promote cell proliferation and HBeAg levels in HepG 2.2.15 cell line, there is a significantly negative correlation between cell proliferation and HBeAg levels. But the both have no correlation with TLR4 expression in vitro. GL could upregulate TLR4 in a dose-independent manner.
出处
《中国现代医学杂志》
CAS
CSCD
北大核心
2010年第4期544-547,共4页
China Journal of Modern Medicine
基金
广东省中医药管理局课题(No:302005)
