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Effect on JNK1 gene silencing on the expression of high-molecular-weight adiponectin in mice with nonalcoholic fatty liver disease
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摘要:
目的探讨JNK1基因沉默对非酒精性脂肪性肝病（NAFLD）小鼠脂肪组织高分子量脂联素（HMW-APN）及相关通路分子的影响。方法 20只C57BL/6小鼠被随机分为正常组、模型对照组、shRNA-JNK1慢病毒处理NAFLD组（JNK1+NAF组）、无关序列shRNA慢病毒处理NAFLD组（无关序列+NAF组）,每组5只。正常饮食与高脂饮食分别喂养3个月,成功复制NAFLD小鼠模型,利用最佳干扰效果的shRNA-JNK1慢病毒和无关序列shRNA慢病毒通过尾静脉注射NAFLD小鼠。根据各组饮食及慢病毒注射情况喂养5 d后,HE染色观察各组小鼠肝组织的病理学改变。ELISA方法检测各组小鼠血清中HMW-APN的水平。取附睾脂肪垫行Western Blot检测分析AMPK、P-AMPK、HMW-APN、DsbA-L的表达水平。计量资料多组间比较采用单因素方差分析,进一步两两比较采用LSD-t检验。结果经HE染色,JNK1+NAF组较模型对照组脂滴空泡减少,水肿、炎症减轻。JNK1+NAF组的肝组织脂肪变评分（2. 267±0. 704）较模型对照组（3. 800±0. 414）和无关序列+...
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关键词:
- 非酒精性脂肪性肝病 /
- 基因沉默 /
- 脂联素 /
- 小鼠,近交C57BL
Abstract:Objective To investigate the effect of JNK1 gene silencing on the expression of high-molecular-weight adiponectin and related pathway molecules in adipose tissue of mice with nonalcoholic fatty liver disease(NAFLD). Methods A total of 20 C57 BL/6 mice were randomly divided into normal group,model control group,NAFLD group treated with shRNA-JNK1 lentivirus(JNK1 + NAF group),and NAFLD group treated with unrelated-sequence shRNA lentivirus(unrelated sequence + NAF group),with 5 mice in each group. Normal diet and high-fat diet were given for 3 months,and a mouse model of NAFLD was successfully established. The mice with NAFLD were given tail vein injection of shRNA-JNK1 lentivirus with the optimal interfering effect and unrelated-sequence shRNA lentivirus. After 5 days of feeding based on diet and lentivirus injection,HE staining was used to observe the pathological changes of liver tissue. ELISA was used to measure the serum level of high-molecular-weight adiponectin. Western blot was used to measure the expression of AMP-activated protein kinase(AMPK),phosphorylated AMPK(p-AMPK),high-molecular-weight adiponectin,and disulfide-bond-A oxidoreductase-like protein(DsbA-L) in epididymal fat pad. A one-way analysis of variance was used for comparison of continuous data between groups,and the least significant difference t-test was used for further comparison between two groups. Results HE staining showed that compared with the model control group,the JNK1 + NAF group had significant reductions in lipid droplet vacuoles,edema,and inflammation. The JNK1 + NAF group had a significantly lower liver steatosis score than the model control group and the unrelated sequence + NAF group(2. 267 ± 0. 704 vs 3. 800 ± 0. 414/3. 667 ± 0. 617,both P < 0. 05). ELISA showed that the JNK1 + NAF group had a significantly higher serum level of high-molecular-weight adiponectin than the model control group(294. 71 ± 102. 30 ng/ml vs 124. 06 ± 70. 58 ng/ml,P<0. 001). Western Blot showed that the JNK1 + NAF group had significantly higher expression levels of AMPK,p-AMPK,DsbA-L,andhigh-molecular-weight adiponectin than the model control group and the unrelated sequence + NAF group(all P < 0. 05). Conclusion JNK1 gene silencing can promote the expression of Dsb A-L and high-molecular-weight adiponectin in NAFLD mice,activate the AMPK pathway to regulate adiponectin multimerization,and thus improve fat deposition in NAFLD.
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Key words:
- nonalcoholic fatty liver disease /
- gene silencing /
- adiponectin /
- mice,inbred C57BL
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