摘要
目的探讨联合应用奥贝胆酸(OCA)和白藜芦醇(RSV)对非酒精性脂肪性肝病(NAFLD)小鼠的治疗效果。方法将37只SPF级C57BL/6Cnc小鼠随机分为空白对照组、NAFLD模型组(MOD组)、OCA组、RSV组、联合用药低剂量组和联合用药高剂量组,除NAFLD模型组7只小鼠外,其余每组6只。应用CCl4和高脂饲料诱导4周建立小鼠NAFLD模型,造模成功后空白对照组和NAFLD模型组灌服等量0.5%CMC-Na溶液,其余各组小鼠灌服0.5%CMC-Na混悬液,剂量分别为OCA组0.04mg/d、RSV组1.33mg/d、联合用药低剂量组0.06mg/d和联合用药高剂量组3.78mg/d。实验全程监测小鼠的体质量变化,每周称重1次,对小鼠用药瘙痒现象进行观察统计并拍照,连续同时给脂给药4周,末次给药8h后,称取小鼠肝脏完整组织湿重测算肝指数。采用贝克曼库尔特AU5800全自动生化分析仪检测小鼠血清生化指标丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)、碱性磷酸酶(ALP)、甘油三酯(TG)、总胆固醇(TC)、低密度脂蛋白胆固醇(LDL-C)和高密度脂蛋白胆固醇(HDL-C)。通过HE染色检测小鼠肝脏组织病理形态学改变。通过油红O染色检测小鼠肝脏组织内脂质分布。采用酶标仪检测10%的小鼠肝组织匀浆液中TG、TC和游离脂肪酸(FFA)的水平。结果与MOD组相比,联合用药低剂量组肝指数减小(P均<0.05);联合用药低剂量组、高剂量组小鼠均出现血清ALT、AST、ALP、TG、TC和LDL-C水平降低(P均<0.05),HDL-C水平均升高(P均<0.05);联合用药低剂量组、高剂量组小鼠肝脏脂肪变性和炎细胞浸润改善,联合用药低剂量组、高剂量组肝脏组织内脂质百分比减少(P均<0.05),联合用药高剂量组百分比减少更显著;联合用药低剂量组、高剂量组小鼠肝脏组织匀浆液中TG和TC含量减少(P均<0.01);联合用药低剂量组、高剂量组小鼠肝脏组织病理学脂肪样变和炎性损伤明显改善。与单独用药组相比,联合用药低剂量组小鼠的肝指数减小(P<0.01),联合用药低剂量组、高剂量组小鼠血清ALT、TG、LDL-C含量水平和LDL-C/HDL-C百分数降低(P均<0.05);联合用药高剂量组小鼠肝脏组织匀浆液中TG和TC含量减少更显著(P均<0.05),联合用药低剂量组、高剂量组小鼠肝脏组织病理学脂肪样变和炎细胞浸润改善更明显,肝脏组织肝内脂滴占比减少更显著(P<0.05)。结论OCA和RSV联用能改善NAFLD小鼠肝功能、降低其血脂水平并改善肝组织病理学变化,有效干预CCl4诱导的高脂饮食小鼠NAFLD发展。
Objective To investigate the therapeutic effect of combined use of obeticholic acid and resveratrol on nonalcoholic fatty liver disease(NAFLD)in mice.Methods Thirty-seven SPF-class C57BL/6Cnc mice were randomly divided into the blank control group(CON group),NAFLD model group(MOD group),obeticholic acid group(OCA group),resveratrol group(RSV group),combined low-dose group(OCA+RSV-L group)and combined high-dose group(OCA+RSV-H group);except the MOD group had 7 mice,there were 6 mice in the other groups.The mouse NAFLD model was established by CCl 4 and high-fat diet for 4 weeks.After the successful modeling,mice in the CON group and MOD group were given the same amount of 0.5%CMC-Na solution,while mice in the other groups were given 0.04 mg/d in the OCA group,1.33 mg/d in the RSV group,0.06 mg/d in the OCA+RSV-L group and 3.78 mg/d in the OCA+RSV-H group for 4 weeks,respectively.The body weight of the mice was monitored throughout the experiment.The rats were weighed once a week.The pruritus of the mice was observed and counted,and the mice were administered for 4 weeks.After the last administration for 8 hours,the intact liver tissues of mice were weighed to measure the liver index.Using Beckman Coulter AU5800 automatic biochemical analyzer to detect the serum biochemical indexes alanine aminotransferase(ALT),aspartate aminotransferase(AST),alkaline phosphatase(ALP),triglycerides(TG),total cholesterol(TC),low-density lipoprotein cholesterol(LDL-C)and high-density lipoprotein cholesterol(HDL-C)in mice.The pathomorphological changes of mouse liver was detected by HE staining.The lipid distribution in the liver tissues of mice was detected by oil red O staining.The levels of TG,TC and free fatty acids(FFA)in 10%mouse liver tissue homogenate were detected by microplate reader.Results Compared with the MOD group,the liver index of the OCA+RSV-L group decreased(P<0.05);the serum levels of ALT,AST,ALP,TG,TC,and LDL-C decreased in both the OCA+RSV-L group and OCA+RSV-H group(all P<0.05),while HDL-C levels increased(P<0.05);liver steatosis and inflammatory cell infiltration were significantly improved in the OCA+RSV-L group and OCA+RSV-H group,and the percentage of lipid in the liver tissues decreased significantly in the OCA+RSV-L group(P<0.05);the percentage in reduction of the OCA+RSV-H group was more significant(P<0.05);the TG and TC levels in the liver tissue homogenate of the OCA+RSV-L group and OCA+RSV-H group decreased(both P<0.01);the histopathological steatosis and inflammation damage of liver in the OCA+RSV-L group and OCA+RSV-H group were significantly improved.Compared with the single-drug group,the liver index of the mice in the OCA+RSV-L group decreased significantly(P<0.01);the serum levels of ALT,TG and LDL-C and percentage of LDL-C/HDL-C in the OCA+RSV-L group and OCA+RSV-H group decreased(all P<0.05);and the decrease of TG and TC in the liver homogenate of the OCA+RSV-H group was more significant(all P<0.05).The histopathological steatosis and inflammation damage in the liver of the OCA+RSV-L group and OCA+RSV-H group were significantly improved,and the percentage of lipid droplets in the liver tissues decreased significantly(both P<0.05).Conclusion The combination of obeticholic acid and resveratrol can improve liver function,lower blood lipid levels and improve histopathological changes in liver tissues,and effectively interfere with the development of NAFLD in mice induced by CCl4.
作者
姜良勇
谷斌
齐亚军
殷俪宁
马伟铭
周奎臣
JIANG Liangyong;GU Bin;QI Yajun;YIN Lining;MA Weiming;ZHOU Kuichen(Jiamusi University,Jiamusi 154007,China)
出处
《山东医药》
CAS
2019年第28期1-6,共6页
Shandong Medical Journal
基金
国家自然科学基金资助项目(21302223)
作者简介
第一作者:姜良勇(1992-),男,硕士,主要研究方向为脂类代谢检测与临床。E-mail:jiangliangyong2019@qq.com;通信作者:周奎臣(1971-),男,硕士,副主任技师,主要研究方向为脂类代谢检测与临床。E-mail:106576993@qq.com.
