实验研究-解毒护肝汤对药物性肝损伤大鼠MMP-2蛋白表达的影响

摘  要:目的  通过检测大鼠血清肝功能指标、血清生物标志物、MMP-2蛋白表达，观察解毒护肝汤对药物性肝损伤大鼠的干预作用及作用机制。方法  将大鼠按体重随机分为正常组、模型组、水飞蓟宾阳性对照组、解毒护肝汤高、中、低剂量组。其中模型组和各给药组给予对乙酰氨基酚药液灌胃复制肝损伤模型，正常组和模型组给予生理盐水灌胃，其余各组给予相应药液灌胃，连续给药30天。实验结束时，检测谷草转氨酶（AST）、谷丙转氨酶(ALT)、总胆红素(T-BIL)、直接胆红素(D-BIL)、苹果酸脱氢酶（MDH）、对氧磷酶（PON1）、嘌呤核苷磷酸化酶（PNP）含量变化以及肝组织基质金属蛋白酶2（MMP-2）的蛋白表达情况。结果：对乙酰氨基酚大鼠血清AST、ALT、DBIL、PON1、MDH、PNP含量均显著升高（p<0.05,p<0.01），解毒护肝汤中、低剂量组能明显降低转氨酶活性及胆红素含量（p<0.05, p<0.01），高剂量组能显著降低肝损伤大鼠血清PON1、MDH含量（p<0.05），中剂量组明显降低MDH的含量（p<0.05）。肝损伤大鼠MMP-2蛋白阳性表达增多；水飞蓟宾对照组以及解毒护肝汤各剂量组阳性表达明显减少。结论：解毒护肝汤对APAP所致的药物性肝损伤具有防治作用。

Abstract: Objective To observe the intervention effect and mechanism of jiedu hugan decoction on drug-induced liver injury in rats by detecting serum liver function indexes, serum biomarkers and MMP-2 protein expression. Methods The rats were randomly divided into normal group, model group, Silybin control group, high dose, medium dose and low dose of jiedu hugan decoction. The model group and each group were given acetaminophen solution by gavage to replicate the liver injury model, the normal group and the model group were given normal saline solution by gavage, and the other groups were given corresponding solution by gavage for 30 consecutive days. At the end of the experiment, we detected the changes of aspartate aminotransferase (AST), alanine aminotransferase (ALT), total bilirubin (T-BIL), direct bilirubin (D-BIL), malate dehydrogenase (MDH), p-oxyphosphatase (PON1), purine nucleoside phosphorylase (PNP) in serum and the protein expressions of matrix metalloproteinase 2( MMP-2) in liver tissue. Results:The serum AST, ALT, DBIL, PON1, MDH, PNP content in rats with given acetaminophen were significantly increased (p < 0.05, p < 0.01), the low dose group of jiedu hugan decoction can obviously decrease transaminase activity and content of bilirubin (p < 0.05, p < 0.01), high dose group can significantly reduce serum PON1, MDH levels (p < 0.05), medium dose group can significantly lower MDH levels (p < 0.05). Positive expression of MMP-2 protein was increased in rats with liver injury. The positive expression of silybin and jiedu hugan decoction in each dose group decreased obviously. Conclusion: Jiedu hugan decoction can prevent and cure the drug-induced liver injury caused by APAP.

对乙酰氨基酚（APAP）及其相关制剂的不合理应用是国外引发急性肝功能衰竭的最主要原因之一[1-2]。对乙酰氨基酚所致的肝损伤可诱导机体内产生一系列的炎性因子变化，进而发生炎症反应[3]。而肝内长期的炎症反应又可促使肝组织不断的修复，在修复过程中细胞外基质（ECM）合成与降解失衡，最终导致肝内结缔组织过量聚集，诱导肝纤维化的发生[4]。由肝细胞膜分泌的基质金属蛋白酶（MMP-2）是ECM代谢过程中的关键酶，可作为早期预测肝纤维化的敏感指标。由于药物性肝损伤的临床表现缺乏特异性，一般需要结合用药史与症状来推测。因此，寻找具有早期诊断功能的血清标志物则显得尤为重要。本文主要探讨具有保肝作用的解毒护肝汤对早期肝损伤指标的调节作用，为解毒护肝中药防治APAP所致药物性肝损伤提供数据支撑。

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解毒护肝汤对药物性肝损伤大鼠MMP-2蛋白表达的影响.pdf