护肝片降低 CCL4肝损伤模型大鼠丙氨酸氨基转移酶作用及其机制

摘要

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150 g/kg为最小有效量；2400 g/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 random-ized into the blank control , model control , CMC control and medication groups with different doses .Hepatic injury was induced by CCL 4 , and activity of serum ALT , AST were tested by colorimetry .Trial of time-effec-tive relationship:Rats were randomized into the blank control , model control , and medication groups with dif-ferent administration time .The dose given to rat was based on the result of dose -effective relationship;hepat-ic injury was induced by CCL 4 , 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 CCL 4 , 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 continu-ously for nine days .The result of mechanism study demonstrated that Huganpian was able to significantly re-duce serum ALT level , and raise liver ALT level , and meanwhile raise mitochondrial transmerabrane potential and the concentration of Ca 2+in hepatocyte .Conclusion:Huganpian is attested to be able to inhibit the activi-ty 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 Ca 2+, Huganpian successfully protects hepatocyte , and induces serum ALT level in rats with decreased releasing .