马氏珠母贝肉提取物对小鼠急性酒精性肝损伤的影响

Improving Effect of Pinctada martensii Meat Extract on Acute Alcoholic Liver Injury in Mice

【目的】探讨马氏珠母贝(Pinctada martensii)肉提取物对小鼠急性酒精性肝损伤(alcoholic liver disease,ALD)的解酒护肝作用。【方法】采用碱溶酸沉法提取贝肉蛋白质,酶解超滤后制备截留分子质量为<3 ku(F-Ⅰ)、3~8 ku(F-Ⅱ)、>8 ku(F-Ⅲ)的超滤组分,同时采用水提法制备马氏珠母贝肉全脏器水提物。以急性酒精性肝损伤小鼠为模型,以小鼠血清生化指标(谷丙转氨酶ALT、谷草转氨酶AST)、肝脏生化指标(超氧化物歧化酶SOD、谷胱甘肽GSH、甘油三酯TG、丙二醛MDA、乙醇脱氢酶ADH和乙醛脱氢酶ALDH)和肝脏病理切片为指标评价各提取物对急性酒精性肝损伤的改善作用,并对抗肝损伤活性较高组分进行氨基酸含量测定及质谱结构鉴定。【结果】与模型组相比,贝肉蛋白酶解超滤组分和水提物组均能极显著降低小鼠血清中ALT和AST活性(P<0.01),降低肝脏中MDA、TG含量(P<0.05),显著提高小鼠肝脏中SOD、GSH含量(P<0.05)、ADH、ALDH活性(P<0.01),改善肝脏组织的病理状况,缓解细胞坏死、炎症等病理状态。氨基酸检测结果显示,F-Ⅰ组分及水提物中氨基酸组成全面,水提物中牛磺酸含量是F-Ⅰ组分的2倍左右。质谱结果表明F-Ⅰ组分、水提物分别包含15、20种主要肽片段。【结论】马氏珠母贝肉蛋白酶解超滤组分和水提物对急性ALD具备保护作用,其中F-Ⅰ组分、水提物效果更佳,其改善机制与增强机体抗氧化能力、促进乙醇代谢能力有关。

【Objective】To futher explore the active anti-alcoholic components in Pinctada martensii meat.【Methods】The ultrafiltration components with a molecular weight of<3 ku(F-I),3~8 ku(F-Ⅱ)and>8 ku(F-Ⅲ)were prepared by enzymatic hydrolysis and ultrafiltration fractionation methods,by using the protein of P.martensii meat extracted by alkali solution and acid precipitation method.Meanwhile,the whole viscera water extract of the shellfish was prepared by the water extraction method.Taking mice with acute alcoholic liver injury as the model,the improvement effect of each extract on the acute alcoholic liver injury was evaluated by serum biochemical indexes(alanine aminotransferase ALT,aspartate aminotransferase AST),liver biochemical indexes(superoxide dismutase SOD,glutathione GSH,triglyceride TG,malondialdehyde MDA,alcohol dehydrogenase ADH and aldehyde dehydrogenase ALDH)and liver pathological sections,and the amino acid content and mass spectrometry structure of components with high activity against liver injury were determined.【Results】Compared with the model group,the ultrafiltration groups and the water extract group could significantly reduce the ALT and AST activities in the serum of mice(P<0.01),reduce the contents of MDA and TG in the liver(P<0.05),and significantly increase the activities of SOD,GSH(P<0.05),ADH and ALDH in the liver of mice(P<0.01),improve the pathological condition of liver tissue,alleviate cell necrosis,inflammation and other pathological conditions.Amino acid detection results showed that the amino acid compositions of F-I fraction and water extract were comprehensive,and the taurine content in the water extract is about twice that of the F-I fraction;mass spectrometry results showed that F-I fraction and water extract contained 15 and 20 main peptide fragments,respectively.【Conclusion】The ultrafiltration fractions and water extract of P.martensii have protective effect on acute ALD,and the F-I fraction and water extract have better effect.The improvement mechanism is related to the enhancement of the antioxidant capacity and promotion of ethanol metabolism.