葡萄籽原花青素对CCl_4诱导小鼠氧化性肝损伤的保护作用及其机制研究

Study on Protective Effect of Grape Seed Proanthocyanidins against Acute Liver Injury Induced by CCl_4 in Mice and Its Mechanism

目的:研究葡萄籽原花青素(GSPE)对小鼠氧化性肝损伤的保护作用及其机制。方法:将24只小鼠随机分为正常组(生理盐水)、模型组(生理盐水)和GSPE低、高剂量组(100、200 mg/kg),连续ig给药4 d,每天1次。末次给药后,除正常组外其余各组小鼠ip含0.2%四氯化碳(CCl4)的植物油溶液复制小鼠肝损伤模型,造模完成后各组小鼠继续ig给药3 d。观察GSPE对急性肝损伤小鼠肝脏大体、病理和肝脏系数的影响;检测血清中丙氨酸转氨酶(ALT)和天冬氨酸转氨酶(AST)活性;检测肝组织中凋亡相关蛋白Bcl-2、Bax和自噬相关蛋白p62的表达;检测肝组织中增殖细胞核抗原(PCNA)蛋白的表达。结果:与正常组比较,模型组小鼠肝脏系数及血清中ALT、AST活性升高;小鼠肝组织全部出现病变(多集中于Ⅲ级);肝组织中Bcl-2、p62蛋白表达减弱,Bax蛋白表达增强,Bcl-2/Bax比值降低(P<0.05或P<0.01),PCNA蛋白表达减弱。与模型组比较,GSPE低、高剂量组小鼠肝脏体积明显缩小,肝细胞坏死和炎症细胞浸润明显减少,病变程度减轻(多集中于Ⅰ、Ⅱ级),肝脏系数明显降低;血清中ALT、AST活性降低;肝组织中Bcl-2、p62蛋白表达增强,Bax蛋白表达减弱,Bcl-2/Bax比值明显升高(P<0.05或P<0.01),PCNA蛋白表达增强。结论:GSPE对CCl4诱导的小鼠氧化性肝损伤具有一定保护作用;其机制可能与GSPE促进小鼠肝细胞增殖、抑制肝组织细胞凋亡和自噬作用有关。

OBJECTIVE：To study protective effect of grape seed proanthocyanidins（GSPE）against acute liver injury in mice and its mechanism. METHODS：24 mice were randomly divided into normal group（normal saline）,model group（normal saline）,GSPE low-dose and high-dose groups（100,200 mg/kg）. They were given relevant medicine intragastrically for consecutive4 days,once a day. After the last administration,those groups were given vegetable oil solution containing 0.2% CCl_4 intraperitoneally to induce liver injury model except normal group;after modeling,those groups continued intragastric administration of medicine for 3 d. The effects of GSPE on liver appearance,pathological and liver coefficient in mice with acute liver injury were observed;the activity of ALT and AST were detected in serum;the expression of apoptosis-related protein Bcl-2 and Bax,that of autophagy-related protein p62 were detected in liver tissue;the expression of PCNA protein in liver tissue was determined. RESULTS：Compared with normal group,liver coefficient,the activity of ALT and AST in serum increased in model group;mice liver tissue pathological changes mostly concentrated in the grade Ⅲ;the expression of Bcl-2 and p62 protein were significantly lowered in liver tissue,while Bax expression was increased,the Bcl-2/Bax was decreased（P〈0.05 or P〈0.01）,the expression of PCNA protein was decreased in liver tissue. Compared with model group,liver volume,necrosis of liver cells and inflammatory cell infiltration decreased significantly,and lesion degree（concentrated in the grade Ⅰ and Ⅱ）and liver coefficient decreased;the activity of ALT and AST in serum decreased;the expression of Bcl-2,p62 and Bcl-2/Bax increased,while protein expression of Bax decreased（P〈0.05 or P〈0.01）,the expression of PCNA protein increased in liver tissue. CONCLUSIONS：GSPE exerts a protective effect on CCl_4 induced oxidative liver injury in mice,and its mechanism may be related to promoting the proliferation of liver cell,inhibiting the apoptosis and autophagy of liver cells.