限时迸食在小鼠和细胞模型中通过溶酶体 生物发生缓解非酒精性脂肪性肝炎

AIM: To investigate the therapeutic effects and molecular mechanisms of time-restricted feeding (TRF)on non-alcoholic steatohepatitis(NASH)mouse liver and oleic acid (0A)-induced damage of human hepatoblastoma HepG2 cells through AMP-activated protein kinase (AMPK)/transcription factor EB (TFEB) -mediated lysosomal biogenesis. METHODS: (1) A mouse NASH model was constructed with a high-fat and high-cholesterol diet. Eighteen C57BL/6J mice were divided into normal control(NC) group, model group, and TRF group, with 6 mice in each group. After 10 weeks of feeding, mice were anesthetized, the eyeballs were removed for blood collection, and the serum was separated to detect the levels of total cholesterol(TC), triglycerides(TG), alanine aminotransferase (ALT), and aspartate aminotransferase （AST） in the serum of the mice. The livers of the mice were collected, and the hepatic coefficients were calculated to detect the hepatic levels of TC and TG. The morphological changes of the livers were observed by hematoxy-lin-eosin（HE）and Masson staining. The protein expression levels of hepatic lysosomal-associated membrane protein 1 （LAMP1）, AMPK, p-AMPK, nuclear TFEB, tumor necrosis factor-cx （TNF-ot）, and interleukin-lp（IL-Ip） were detected by Western blot method. （2）HepG2 cells were intervened with OA to establish a liver injury model and serum deprivation was used to simulate fasting conditions. HepG2 cells were divided into control, serum deprivation（FBS-）, OA, and OA+FBS-groups. siRNA knockdown of TEEB was used to investigate the relationship between TFEB-mediated lysosomal biogenesis and hepatocyte lipid accumulation and liver injury. AMPK inhibitor compound C （CC） was used to inhibit AMPK activity to study the relationship between AMPK and TFEB-mediated lysosomal biogenesis. Hepatocyte lipid accumulation was detected by oil red 0 staining. Hepatocyte TC, TG, ALT, and AST levels were detected by kits. Hepatocyte LAMP1, AMPK, p-AMPK, and nuclear TFEB protein expression levels were detected by Western blot. RESULTS; （ 1） TRF intervention significantly reduced serum TC, TG, ALT, and AST levels and the expression of IL-lp, TNF-ot, TG, and TC in the liver of NASH mice, and alleviated hepatic steatosis and inflammatory infiltration（PV0. 05）. （2）Serum deprivation intervention reduced the number of lipid droplets, as well as TG, ALT, and AST levels of OA-induced HepG2 cells （PVO. 01）. （3）Western blot results showed that TFEB nuclear translocation level, LAMP1 protein levels, and AMPK phosphorylation levels were significantly increased in the liver of NASH mice after TRF intervention（PV0. 01）. While TFEB nuclear translocation level, LAMP1 protein level, and AMPK phosphorylation level were significantly increased in OA-induced HepG2 cells after serum deprivation intervention （f<0. 01）. （4）After the siRNA intervention, TFEB and LAM Pl protein levels were decreased, and the ameliorative effects of serum deprivation on lipid accumulation and liver injury in the OA group were significantly attenuated（P<O. 05）. （5）After CC intervention, the AMPK phosphorylation level, LAMP1 protein level, and TFEB nuclear translocation level were significantly reduced （P<0. 01）. CONCLUSION: （1） TRF can alleviate lipid accumulation and inflammation in the liver of NASH mice. （2）The beneficial effects of TRF on the liver of NASH mice may be related to its contribution to AMPK/TFEB-mediated lysosomal biogenesis. [ABSTRACT FROM AUTHOR]