鲟鱼鱼油的酶法提取及对非酒精性脂肪肝病的干预作用

Enzymatic extraction of sturgeon fish oil and its intervention on non-alcoholic fatty liver disease

目的优化酶法提取鲟鱼鱼油工艺,探讨其对非酒精性脂肪肝病(non-alcoholic fatty liver disease,NAFLD)的干预作用。方法以鲟鱼下脚料为原料,用酶法提取鱼油,在单因素实验的基础上,采用正交实验得到提取鲟鱼鱼油的最佳工艺。利用气相色谱-质谱法检测鱼油的脂肪酸组成及含量。采用高脂饲喂方法建立NAFLD小鼠模型,探究鲟鱼鱼油对小鼠NAFLD的改善作用。结果鲟鱼鱼油最佳提取条件为:1500 U/g木瓜蛋白酶、料液比1:1(mL/g)、pH 7、45℃酶解时间2 h。此条件鱼油的提取率为82.94%,所得鱼油为淡黄色、澄清透明,酸价为0.1 mg KOH/g、过氧化值为0.49 mmol/kg,符合一级鱼油国家标准。提取的鲟鱼鱼油饱和脂肪酸为23.56%、不饱和脂肪酸为74.71%。动物实验结果显示,正常饲料中添加鲟鱼鱼油时,小鼠各项指标均无显著性变化;高脂组小鼠血清中血清总胆固醇、甘油三酯、低密度脂蛋白、丙氨酸氨基转移酶、天冬氨酸氨基转移酶显著升高,高密度脂蛋白显著降低,而高脂组中添加深海鱼油和鲟鱼鱼油可显著改善血脂异常;添加鲟鱼鱼油可明显改善由高脂饲喂引起的小鼠肝组织脂肪变性及炎性浸润;添加鲟鱼油可显著改善小鼠肝组织炎性细胞因子基因IL-1β、TNF-α、IL-6、MCP1,脂代谢有关基因CD36、SREBF1及胆固醇代谢相关基因HMGCR、CYP7A1的表达。结论本研究优化了鲟鱼鱼油提取工艺,获得的鲟鱼鱼油中ω-3不饱和脂肪酸含量较高,证明了鲟鱼鱼油对NAFLD的改善作用,可作为深海鱼油的替代产品。

Objective To optimize the enzymatic extraction process of sturgeon fish oil,and explore its interventional effect on non-alcoholic fatty liver disease(NAFLD).Methods Using sturgeon scraps as raw materials,the fish oil was extracted by enzyme method,the optimal process for extracting sturgeon fish oil was obtained by orthogonal experiment on the basis of single factor experiment.The composition and content of fatty acids were detected by gas chromatography-mass spectrometry.The mouse model of NAFLD was established by feeding a high-fat diet to study effect of sturgeon fish oil on improving NAFLD in model mice.Results The optimal extraction conditions of sturgeon fish oil were 1500 U/g papain,the ratio of solid to liquid 1:1(mL/g),pH 7,and the enzymatic hydrolysis time at 45℃for 2 h.The extraction rate of fish oil under this condition was 82.94%,the fish oil extracted under this condition was light yellow,clear and transparent,the acid value was 0.1 mg KOH/g,and the peroxide value was 0.49 mmol/kg,which met the national standard for first-class fish oil.The saturated fatty acids and unsaturated fatty acids of extracted sturgeon oil were 23.56%and 74.71%respectively.The results of animal experiments showed that when sturgeon fish oil was added to the normal feed,there were no significant changes in various indicators in mice;serum total cholesterol,triglycerides,low-density lipoprotein,alanine transferase and aspartate transferase significantly increased,and high-density lipoprotein significantly decreased in high-fat group,adding deep-sea fish oil and sturgeon oil could significantly improve dyslipidemia;the addition of sturgeon oil significantly improved the fatty degeneration and inflammatory infiltration of liver tissue caused by high-fat feeding,the addition of sturgeon oil significantly improved the expression of inflammatory cytokines TNF-α,IL-1β,IL-6,MCP1,lipid metabolism related genes CD36,SREBF1 and cholesterol metabolism related genes HMGCR,CYP7A1.Conclusion In this study,the extraction process of sturgeon fish oil has been optimized,and the obtained sturgeon fish oil has a higher content ofω-3 unsaturated fatty acids,and the improvement effect of sturgeon fish oil on NAFLD has been proved,and sturgeon fish oil can be used as an alternative product of deep-sea fish oil.