灵芪蠲肝胶囊含药血清对活化大鼠肝星状细胞凋亡的影响

Effects of LingQiJuanGan Capsule Medicated Serum on Apoptosis of Activated Rat Hepatic Stellate Cells

目的观察灵芪蠲肝胶囊含药血清对血小板衍化生长因子(PDGF)诱导的活化大鼠肝星状细胞(HSC-T6)凋亡的作用,并观察其对凋亡基因Bcl-2、Bax蛋白表达的影响,探讨其可能的抗肝纤维化机制。方法首先制备含药血清:SD大鼠随机分为3组,分别用生理盐水(A组,10mL/kg),复方鳖甲软肝片药液(B组,1.5g/kg),灵芪蠲肝胶囊药液(C组,4.25g/kg)灌胃7d,每日1次。经腹主动脉采血,静置、离心、灭活、过滤,取得含药血清。以PDGF 10ng/mL刺激活化HSC-T6。将各组含药血清按200mL/L分别作用于HSC-T6。0、12、24、48h后采用流式细胞技术检测各组HSC-T6凋亡情况以及Bcl-2、Bax的表达。结果 B、C组含药血清作用于HSC-T6细胞12、24、48h,细胞凋亡率较A组升高,差异有统计学意义(P<0.05),而B、C组间的差异无统计学意义(P>0.05)。各组组内不同时间点比较,除A组外,B、C组在不同时间点的两两比较差异均有统计学意义,细胞凋亡率随时间的延长出现明显上升(P<0.05)。含药血清作用于HSC-T6细胞12、24、48h,B组、C组Bcl-2的表达较A组明显下降,Bax的表达出现明显上升,差异均有统计学意义(P<0.05),而B组与C组间差异均无统计学意义(P>0.05)。除A组外,B组和C组在不同时间点的两两比较差异均有统计学意义(P<0.05),Bcl-2蛋白的表达在0h最高,随时间的延长表现为下降趋势(P<0.05),Bax蛋白的表达随着时间的延长,整体表现为上调趋势(P<0.05)。结论灵芪蠲肝胶囊可以诱导活化的HSC-T6凋亡,其机制可能为影响Bcl-2/Bax表达失衡,从而发挥其抗纤维化作用。

Objective To investigate the effects of LingQiJuanGan Capsule medicated serum on the apoptosis of and the expression of Bcl-2/Bax in hepatic stellate cell-T6（HSC-T6）activated by platelet-derived growth factor（PDGF）.Methods Preparation of drug-medicated serum.30SD rats were randomly divided into three groups.Serum was obtained from the SD rats administrated intragastricly with saline（10mL/kg,group A）,Fufang Biejia Ruangan Tablet solution（1.5g/kg,group B）and LingQiJuanGan Capsule solution respectively（4.25g/kg,group C）.Detection of cell apoptosis and the expression of Bcl-2/Bax.HSC-T6cells were activated by 10ng/mL PDGF and incubated with drug-medicated serum at 200 mL/L.Cell apoptosis and the expression of Bcl-2/Bax were measured by flow cytometry at 0,12,24and 48h.Results Significantly higher apoptotic rates of HSC-T6in group B and group C were detected at 12,24,48hcompared with that of group A（P＆lt;0.05）,wheraus no significant difference was found between group B and group C（P＆gt;0.05）.With increase of exposure time,the apoptotic rates of HSC-T6in group B and group C increased（P＆lt;0.05）.The levels of Bcl-2expression in HSC-T6in group B and C were lower than that of group A（P＆lt;0.05）at 12,24,48h（No significant difference between group B and group C,P＆gt;0.05）.The expression level of Bcl-2decreased with increased exposure time（P＆gt;0.05）in both group B and group C.However,no such trend was observed in group A（P＆gt;0.05）.In both group B and group C,the expression level of Bax increased in a time-dependent manner.Conclusion LingQiJuanGan Capsule can induce apoptosis of PDGF activated HSC-T6and inhibit hepatic fibrosis in a time-dependent manner by influencing the expression of Bcl-2/Bax.