中医寒气对高脂饮食诱导肥胖模型小鼠脂肪组织TRPM8通路的影响

Effects of Cold Qi on TRPM8 Pathway in Adipose Tissue of Obese Mice Induced by High Fat Diet

目的通过观察中医寒气对高脂饮食诱导肥胖型小鼠脂肪组织TRPM8的影响,探讨寒气减肥效应的作用机制。方法30只6~8周龄的雄性C57BL/6J野生型小鼠和TRPM8^(-/-)小鼠随机分为3组,分别为C57BL/6J 30℃组、C57BL/6J 4℃组、TRPM8^(-/-)4℃组。采用高脂饲养的方法复制肥胖型小鼠模型。所有小鼠首先饲养在温度为30℃、相对湿度为(55±15)%、室内风速0.1~0.2 m/s、光照周期12 h的笼子中4周,然后将C57BL/6J 4℃组、TRPM8^(-/-)4℃组小鼠转移至18℃的温度下适应性饲养1周,接着转移至4℃的低温饲养箱中饲养4周。作为对照组,C57BL/6J 30℃组小鼠始终在30℃环境下饲喂。9周后,取各组小鼠肩胛、附睾、腹膜脂肪组织并用HE染色法、免疫组织化学染色及Western blot法检测其TRPM8蛋白的表达。结果与C57BL/6J 4℃比较,TRPM8^(-/-)4℃组和C57BL/6J 30℃组肩胛和腹膜脂肪细胞数量显著减少(P<0.001),TRPM8蛋白的表达明显减少。结论中医寒气可诱导肥胖小鼠脂肪细胞TRPM8蛋白的高表达,影响脂肪代谢,有望成为减肥药物研发的新策略,TRPM8有望成为减肥药物研发的新靶点。

Objective To observe the effect of traditional Chinese medicine cold Qi on adipose tissue transient receptor potential M8(TRPM8)in obese mice induced by high fat diet,and to explore the mechanism of the effect of cold Qi on weight loss.Methods Thirty male C57BL/6J wild-type mice and TRPM8^(-/-)mice aged 6-8 weeks were randomly divided into 3 groups:C57BL/6J 30℃group,C57BL/6J 4℃group and TRPM8^(-/-)4℃group.High fat feeding method was used to replicate the obese mouse model.All mice were first reared in a cage with a temperature of 30℃,relative humidity of(55±15)%,indoor wind speed of 0.1-0.2 m/s and light cycle of 12 h for 4 weeks.Then the mice in C57BL/6J group 4℃and TRPM8^(-/-)4℃groups were transferred to a temperature of 18℃for 1 week.They were then transferred to a low temperature feeding box at 4℃for 4 weeks.As the control group,the mice in C57BL/6J 30℃group were fed at 30℃all the time.After 9 weeks,the adipose tissue of shoulder,epididymis and peritoneum of mice in each group was collected and the expression of TRPM8 protein was detected by HE staining,immunohistochemical staining and Western blot.Results Compared with those of the C57BL/6J 4℃group,the shoulder and peritoneal fat cells of the TRPM8^(-/-)4℃and C57BL/6J 30℃groups significantly reduced(P<0.001)and TRPM8 protein expression decreased significantly.Conclusion Traditional Chinese medicine cold Qi can induce the high expression of TRPM8 protein in fat cells of obese mice and affect the fat metabolism,which is expected to be a new strategy for the development of slimming drugs,and TRPM8 is expected to be a new target for the development of slimming drugs.