凉血化瘀方抗肝细胞凋亡效应及对半胱天冬酶-3 mRNA表达的影响

目的研究半胱天冬酶-3(caspase-3)mRNA在内毒素肝损伤中的表达与肝细胞凋亡的关系,以及凉血化瘀方对其的影响.方法腹腔注射内毒素脂多糖(LipoplySaccharide,LPS)于D-氨基半乳糖(D-galactosamine,GalN)致敏的小鼠,造成暴发性肝衰竭(Fulminant Hepatic Failure,FHF)模型,用逆转录-聚合酶链反应(RT-PCR)半定量分析caspase-3 mRNA表达,脱氧核苷酸转移酶介导的缺口原位末端标记技术(TUNEL法)、HE染色及透射电镜观察肝细胞凋亡情况,同时观察药物的影响作用.结果造模3h肝组织caspase-3 mRNA明显表达,治疗组83%,模型组141%,6h二组表达水平接近.凋亡阳性细胞在6h表达最强,二组比较治疗组则明显减少(P＜0.01).结论内毒素肝损伤中caspase-3 mRNA表达与肝细胞凋亡程度有相关关系,凉血化瘀方能延缓内毒素肝损伤肝细胞caspase-3 mRNA表达和抑制其凋亡效应.

Anti-hepatocytic Apoptosis-effect of the Formula Functioning in Cooling Blood to Remove Blood Stasis and the Formula''''s Influence to the Expression of Caspase-3 mRNA

Objective To study the expression of caspase-3 mRNA in hepatic impairment induced by LPS and the relationship between which and hepatocytic apoptosis, and to study the effect of the formula with the function of cooling blood to remove blood stasis. Method FHF was induced by injection of LPS into sensited mice by D-galactosamine (GalN). To analyze the expression of caspase-3 mRNA by using RT-PCR semi-quantitative technique. The condition of hepatocytic apoptosis was examined by TUNEL mediated by deoxyribonucleotide transferring enzyma, HE staining and transmission electron microscopy. Meanwhile, to observe the medicine's function. Result Obvious expression of caspase-3 mRNA in the model mice in three hours 141% and the treatment group 83%, in six hours, the expressions of the two groups were very close. The positive apoptotic cells expressed most strongly in the model mice in six hours, while comparatively speaking, the treatment group was obviously reduced (P<0.01). Conclusion The expression of caspase-3 mRNA in hepatic impairment induced by LPS was related to the condition of hepatocytic apoptosis. The formula functioning in cooling blood to remove blood stasis can delay the expression of caspase-3 mRNA in hepatocytic apoptosis induced by LPS and inhibit hepatocytic apoptosis.