蕲艾提取液含药血清对肝星状细胞转化生长因子β_1及Smad蛋白表达的影响

Effect of Artemisia Argyi Extracting Solution Contained Rat Serum on Eexpression of Transforming Growth Factor-β_1 and Protein Smads in Hepatic Stellate Cells

目的探讨蕲艾提取液的抗肝纤维化作用及其机制。方法 24只Wistar大鼠随机分为蕲艾组、丹参组、生理盐水组和空白组,每组6只。其中,蕲艾组用20%的蕲艾提取液灌胃,丹参组用丹参注射液灌胃,3组灌液量均为20ml/kg,每天3次,连续3天,末次给药1h后采血取血清,用细胞培养液DMEM配制成体积分数分别为5%、10%和20%的含药培养液。分别检测各体积分数的蕲艾提取液含药培养液对HSC/T6细胞转化生长因子(TGF-β1)、Smad3、Smad7mRNA和蛋白的影响。结果 5%、10%和20%的蕲艾提取液含药血清培养HSC/T6细胞24h后TGF-β1mRNA表达水平分别为空白组的(0.82±0.03)、(0.72±0.03)和(0.67±0.04)倍,蛋白表达水平分别为(0.66±0.09)、(0.34±0.05)和(0.31±0.07)倍,与生理盐水组相比显著降低(P<0.05);且均下调Smad3mRNA和蛋白水平,同时上调Smad7mRNA和蛋白表达。蕲艾提取液含药血清20%组与丹参组相比无统计学差异。结论蕲艾提取液含药血清抑制肝星状细胞TGF-β1和Smad3的表达,上调Smad7的表达,从而抑制HSC/T6活化、增殖,降低胶原的表达及分泌,达到抗纤维化的目的。

Objective To investigate the effect and mechanism of Artemisia Argyi extracting solution on anti-hepatic fibrosis.Methods Twenty-four experimental Wistar rats were randomized into four groups：Artemisia group,Danshen group,physiological saline group,and blank control group,with 6 in each.The Artemisia group was treated with 20% Artemisia Argyi extracting solution,the Danshen group with Danshen （salvia） solution,and the physiological saline group with physiological saline by gavage,20ml/kg.After treating three times per day for 3 days,sera were obtained and made a mixture with DMEM to be volume fraction as 5%,10%,and 20%.Effect of different volume fraction of extracting solution on TGF-β1 Smad3 and Smad7 mRNA and protein were detected.Results After treating with 5%,10% and 20% Artemisia Argyi extracting solution contained rat serum for 24h,contrasted to the black control group,the expression of TGF-β1 mRNA level was （0.82±0.03）,（0.72±0.03） and （0.67±0.04） times,and the expression of TGF-β1 protein was （0.66±0.09）,（0.34±0.05） and （0.31±0.07） times respectively;contrasted to the physiological saline group,the expressions of TGF-β1 mRNA and protein were significantly decreased （P0.05）,and the expressions of Smad3 mRNA and protein were down-regulated and at the same time the expressions of Smad7 mRNA and protein were up-regulated.There was no significant difference between the 20% Artemisia Argyi extracting solution contained rat serum group and the Danshen group.Conclusion Artemisia Argyi extracting solution contained rat serum plays a role in anti-fibrosis by the way of reducing TGF-β1 and Smad3 and up-regulating Smad7,which leads to inhibiting the activation and proliferation of HSC-T6 to reduce the collagen expression and secretion.