刺络泻血疗法对四氯化碳诱导的HF大鼠PCⅢ、CⅣ的影响

摘要

目的:本实验采用用四氯化碳(CCL4)诱导的SD大鼠HF模型,在大鼠足三里、太冲、阳陵泉这3个穴位上进行刺络泻血治疗,观察该疗法对HF模型大鼠肝脏组织形态、肝功能AST、ALT及肝纤维化指标PCⅢ、CⅣ的影响,从而探索刺血疗法对于HF的疗效及其作用机制.方法:Sprague-Dawley(SD)大鼠40只,雄性,随机分为4组,空白对照组(10只)、模型对照组(10只)、秋水仙碱组(10只)以及刺血干预组(10只).除空白对照组外,其余3组用40％CCL4(花生油稀释)诱导HF模型,为期6周.第6周至第12周,对空白对照组的大鼠0.9％生理盐水皮下注射,2次/周,至12周;其余3组的大鼠持续注射40％CCL4(花生油稀释),模型对照组的大鼠只抓取,不做其他处理,秋水仙碱组同时220μg/(kg·d)标准给予秋水仙碱,5次/周;刺络泻血组同时进行刺络泻血疗法干预,2次/周,至12周.12周后,麻醉大鼠并行开腹术,取大鼠肝左叶小块组织行石蜡包埋、切片,以及Masson染色,观察大鼠肝左叶的组织形态学改变;采用ELisa法对肝纤维化指标PCⅢ、CⅣ,采用化学法对肝功能指标AST、ALT进行定量检测.结果:1)肝脏组织形态改变:对切片分析后可见生理盐水对照组肝小叶结构完整,汇管区周围存在少量染色;模型组肝小叶消失,纤维架桥出现,形成肝纤维化;秋水仙碱组和刺血干预组肝纤维化程度明显比造模组减轻,但比空白对照组严重,说明秋水仙碱组和刺血干预组干预均有效.2)肝功能情况:比较于模型对照组,刺血干预组AST、ALT含量显著降低(P<0.05),秋水仙碱组AST含量降低(P>0.05),而ALT含量显著降低(P<0.05);提示刺血疗法可显著降低血清AST、ALT的含量.3)对肝纤维化指标PCⅢ、CⅣ的影响:比较于模型对照组,秋水仙碱组和刺血干预组PCⅢ、CⅣ的含量下降,差异有统计学意义(P<0.05)提示刺血疗法可以显著降低PCⅢ、CⅣ的含量.%Objective:To observed effect of the pathological change of the liver , liver function and apoptosis factor PCIII and PCIV in HF model rats induced by CCL 4, and with the treatment on or near Taichong , Zusanl and Yanglingquan of the rats by pricking blood therapy with a three-edged needle , so as to explore the mechanism of bloodletting in prevention and treatment of HF rats . Methods:Forty male SD rats were randomly divided into normal control group , model group , colchicines-controlled group and bloodletting group with 10 cases in each group .Except the normal control group , the other three groups were induced into liver fi-brosis model with 40％CCL4 diluted by peanut oil .After 6 weeks, the colchicines-controlled group was given colchicine 200μg/( kg· d) with 5 times a week; the blood-letting group had 0.15-0.2 mL hemorrhage on or near Taichong , Zusanl and Yan-glingquan with 2 times a week.After 12 weeks, the general conditions of rats were observed;Small piece of liver tissue selected from left lobe of liver was embedded in paraffin , sliced and stained with Masson and its pathological changes were observed ;The liver function AST , ALT were detected by ELISA method;PCⅢand PCIV were detected by the expression by fluorescence semi quantitative PCR .Results:The morphological change of liver tissue:Masson staining showed structural integrity of hepatic lobule . There were only a little collagen fibers in portal area .Compared with normal group , masson staining result indicated significant liv-er fibrillation .The fibrillation of colchicines-controlled group and bloodletting group was lighter than that in model group , indica-ting the treatment was effective .Liver function:compared with the model group , AST, ALT levels of the pricking blood group de-creased significantly (P<0.05), and AST content of colchicine control group decreased (P>0.05).And the content of ALT sig-nificantly reduced ( P<0.05);Pricking blood therapy could significantly reduce the serum AST and ALT levels;PCIII, CIV ex-pression:compared with the model group , PC III, C IV expression of colchicine group and thorn blood group decreased , and the difference had statistical significance ( P<0.05 ) .The pricking blood therapy can significantly down regulated the expression of PCIII and CIV.Conclusion:Firstly, bloodletting therapy can reduce the content of serum ALT and AST to a certain extent , and can protect liver function .Secondly, bloodletting therapy can significantly reduce the expression of PCIII and CIV .Thirdly, blood-letting therapy is effective in the treatment of liver fibrosis in rats , which can reduce the expression of apoptosis inhibiting gene PCIII and CIV, as well as change the ratio of PCIII/CIV.