自噬相关基因在肥胖患者及肥胖小鼠脂肪组织中的研究

Activated autophagy and inflammation of adipose tissue is involved in obesity

目的:观察肥胖患者及肥胖小鼠脂肪组织中自噬相关基因的变化情况,初步探讨自噬在肥胖中的作用。方法:将C57雄性小鼠随机分为正常饮食组(n=6)及高脂饮食组(n=6),分别给予正常或高脂饮食喂养8周后处死小鼠,收集其附睾及腹股沟脂肪组织。另收集年龄及性别匹配的非肥胖与肥胖患者大网膜脂肪组织。RT-PCR法检测人脂肪组织中自噬相关基因Beclin、ATG1的表达。Western blot方法检测小鼠与人脂肪组织中自噬相关基因LC3、P62的表达水平。结果:1高脂组小鼠体质量、腹股沟脂肪质量/体质量、附睾脂肪质量/体质量均较对照组显著增加,提示肥胖小鼠模型构建成功;2与正常饮食组相比,高脂饮食小鼠脂肪组织中LC3蛋白表达明显增高(t=2.638,P=0.020),P62蛋白水平降低(t=2.264,P=0.047);3肥胖患者脂肪组织中Beclin(t=1.933,P=0.029)、ATG1(t=0.864,P=0.039)的m RNA表达水平及LC3(t=2.847,P=0.015)蛋白表达水平均较正常者明显增高,而P62(t=2.571,P=0.026)蛋白水平显著降低。结论:在肥胖小鼠与肥胖患者脂肪组织中,自噬基因表达明显改变,提示自噬可能参与了肥胖相关的脂肪代谢紊乱。

Objective：To investigate the autophagy and inflammatory cytokine expression of adipose tissue in high-fat diet induced obese mice and obesity human. Methods：Sixteen male C57 mice were randomly divided into chow diet（CD,n=6）group and high-fat diet（HFD,n=6）group. Mice epididymal and inguinal adipose tissues were collected. Omental adipose tissue was obtained from lean（n=9）and obese（n=9）subjects undergoing laparoscopic cholecystectomy;the two groups of participants were age-and sex-matched,with available clinical parameters. The m RNA levels of Beclin and ATG1 were determined by RT-PCR. Western blot was used to detect the protein expression levels of LC3 and P62 in adipose tissue of each sample. Results：After 8 weeks,mice in HFD group displayed higher weight than control group. Compared with control group,HFD-induced obese mice displayed significant higher protein levels of LC3 and significant lower level of P62 in adipose tissue. The m RNA levels of Beclin（P=0.029）and ATG1（P=0.039）were significant higher in the obesity human adipose tissue than in the lean group. The protein expression levels of LC3 were also significantly higher in obesity group than in control group（P=0.015）. Conclusion：Autophagy could be involved in obesity-related adipose tissue dysfunction.