世界中医药

作者	单位
张博1，赵慧玲1，谭丽2，魏珂3，魏丹蕾1，曾蕊1	1 北京中医药大学针灸推拿学院,北京,100029 2 广州中医药大学,广州,510405 3 北京丰台中西医结合医院,北京,100071

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中文摘要:

目的:本实验采用用四氯化碳(CCL4)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% CCL4(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% CCL4(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220 μg/（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。

English Summary:

To observed effect of the pathological change of the liver, liver function and apoptosis factor PCIII and PCIV in HF model rats induced by CCL4, 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 fibrosis model with 40% CCL4 diluted by peanut oil. After 6 weeks, the colchicines-controlled group was given colchicine 200 μg/（kg·d） with 5 times a week; the blood-letting group had 0.15-0.2 mL hemorrhage on or near Taichong, Zusanl and Yanglingquan 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; PCⅢ and 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 liver fibrillation. The fibrillation of colchicines-controlled group and bloodletting group was lighter than that in model group, indicating the treatment was effective. Liver function:compared with the model group, AST, ALT levels of the pricking blood group decreased significantly (P<0.05), and AST content of colchicine control group decreased (P>0.05). And the content of ALT significantly reduced (P<0.05); Pricking blood therapy could significantly reduce the serum AST and ALT levels; PCIII, CIV expression: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, bloodletting 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.

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