摘要
目的研究苦参碱对体外培养的大鼠肝星状细胞系rHSC-99凋亡的影响,并探讨其机制。方法体外培养rHSC-99,随机分成4组:对照组(A组),苦参碱0.12 mg/kg组(B组),苦参碱0.24 mg/kg组(C组),苦参碱0.48 mg/kg组(D组)。苦参碱作用24 h后,TUNEL法检测rHSC-99凋亡,半定量RT-PCR方法及免疫细胞化学方法检测rHSC-99中神经生长因子(NGF),p75及Caspase-3的表达情况。结果不同浓度的苦参碱干预rHSC-99 24 h后,rHSC-99凋亡率随苦参碱的浓度增大而增高,D组凋亡率最高(P<0.01);NGF,p75,Caspase-3mRNA及其蛋白表达随苦参碱的浓度加大而表达升高,C组表达最高(P<0.01)。结论苦参碱可诱导HSCs凋亡。激活NGF/p75通路、上调Caspase-3蛋白表达可能是苦参碱诱导肝星状细胞凋亡的机制之一。
Objective To investigate the effect of matrine on inducing apoptosis of rat hepatic stellate cell line-99（r HSC-99） in vitro and to decipher the underlying mechanism. Methods rHSC-99 cells were treated with matrine of different concentration of 0.12,0.24 and 0.48 mg/mL for 24 h respectivdy. The induced apoptosis of HSC was detected by Terminal deoxynucleotidy transferase mediated dU TP nick end labeling（TUNEL）. RT-PCR and immuocytochemistry were used to detect the mRNA and protein level of NGF, p75, and Caspase-3 respectively. Results Administration of 0.12-0.48 mg/mL matrine could significantly induce r HSC-99 apopto sis in a do se-dependent manner with most cells going apopto sis in 0.48 mg/mL（P 〈 0.01）. mRNA and protein exp re ssio n levels of N GF, p75 and Caspase-3 in rH- SC-99 cells treated with different concentration of matrine were also increasedin a dose-dependent manner, with highest ones detected by the administration of 0.24 mg/mL of matrine P 〈 0.01）. Conclusion Matrine can significantly induce the apoptosis of rHSC-99 cells. Activation of the NGF/p75 pathway and upregulation of Caspase-3 expression may be one of the underlying mechanisms of rH SC-99 cell apoptosis induced by matrine.
出处
《福建医科大学学报》
2009年第6期443-446,共4页
Journal of Fujian Medical University
基金
福建省自然科学基金(C0540011)
