护肝片降低CCL_4肝损伤模型大鼠丙氨酸氨基转移酶作用及其机制

Mechanism of Huganpian for Inhibiting the Activity of Alanine Aminotransferase in CCL-induced Hepatic Injury in Rats

目的:探讨护肝片降低CCL4肝损伤大鼠血清丙氨酸氨基转移酶(ALT)的作用特点及内在机制。方法:量效关系试验:将昆明种小鼠随机分为空白组、模型对照组、CMC对照组及不同药物剂量组,采用四氯化碳造模,比色法测定血清ALT、AST活性。时效关系试验:将昆明种小鼠随机分为空白组、模型组及不同给药时间组。根据量效关系研究结果,各给药组给药剂量为最大有效量,采用四氯化碳造模,比色法测定血清中AST、ALT的活性。将Wistar雄性大鼠随机分为空白对照组、模型对照组及护肝片治疗组,根据量效及时效关系试验结果给药,采用四氯化碳造模,比色法测定血清及肝组织ALT活性,荧光分光光度法测定肝细胞线粒体膜电位及肝细胞内钙离子浓度。结果:量效试验结果表明,护肝片150g/kg为最小有效量;2400g/kg为最大有效量。时效试验结果表明,护肝片给药3天为起效时间;给药9天达到最大药效。机制试验结果显示,护肝片组能显著降低CCL4肝损伤大鼠血清ALT水平,升高肝组织ALT水平,升高肝细胞线粒体膜电位,降低肝细胞内游离钙离子浓度。结论:在该实验条件下,护肝片具有降低CCL4诱导肝损伤模型大鼠血清ALT活性的作用,并确定了量效及时效关系。护肝片是通过保护线粒体,降低细胞内钙离子浓度,从而保护肝细胞,抑制肝细胞ALT的释放,进而降低模型大鼠血清ALT水平。

Objective： To observe the property and inner mechanism of Huganpian for inhibiting the activity of ALT in CCL-induced hepatic injury in rats. Methods： Trial of dose-effective relationship： Rats were randomized into the blank control,model control,CMC control and medication groups with different doses. Hepatic injury was induced by CCL4,and activity of serum ALT,AST were tested by colorimetry. Trial of time-effective relationship： Rats were randomized into the blank control,model control,and medication groups with different administration time. The dose given to rat was based on the result of dose-effective relationship; hepatic injury was induced by CCL4,and activities of serum ALT,AST were tested by colorimetry. Wistar rats were randomized into the blank control,model control,positive control and medication groups. Administration dose and time were based on the results of dose-effective relationship and time-effective relationship; and hepatic injury was induced by CCL4,and activities of serum and liver ALT were tested by colorimetry. Mitochondrial transmerabrane potential and the concentration of Ca2 ＋in hepatocyte were both tested by spectrophotometry.Results： Trial of dose-effective relationship indicated that the minimum effective dose was 150g /kg,and the maximum effective dose was 2400g /kg. Trial of time-effective relationship indicated that Huganpian had the effect when administrated continuously for three days,and the effect maximized when administrated continuously for nine days. The result of mechanism study demonstrated that Huganpian was able to significantly reduce serum ALT level,and raise liver ALT level,and meanwhile raise mitochondrial transmerabrane potential and the concentration of Ca2 ＋in hepatocyte. Conclusion： Huganpian is attested to be able to inhibit the activity of ALT in CCL-induced hepatic injury in rats in this experiment,of which the dose-effect plus time effect relationships are confirmed. By protecting mitochondrion and reducing intracellular concentration of Ca2 ＋,Huganpian successfully protects hepatocyte,and induces serum ALT level in rats with decreased releasing.