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ObjectiveThis study was undertaken to detect if free fatty acids (FFA) induce hepatocyte senescence in L-02 cells and if huperzine A has an anti-aging effect in fatty liver cells.MethodsL-02 cells were treated with a FFA mixture (oleate/palmitate, at 3:0, 2:1, 1:1, 1:2 and 0:3 ratios) at different concentrations. Cell viability and fat accumulation rate were assessed by a Cell Counting Kit 8 and Nile Red staining, respectively. The mixture with the highest cell viability and fat accumulation rate was selected to continue with the following experiment. The L-02 cells were divided into five groups, including the control group, FFA group, FFA + 0.1 μmol/L huperzine A (LH) group, FFA + 1.0 μmol/L huperzine A (MH) group and FFA + 10 μmol/L huperzine A (HH) group, and were cultured for 24 h. The expression of senescence-associated β-galactosidase (SA-β-gal) was detected by an SA-β-gal staining kit. The expression levels of aging genes were measured by qRT-PCR. The expression levels of apoptosis proteins were detected by a Western blot. ELISA kits were used to detect inflammatory factors and oxidative stress products. The expression of nuclear factor (NF-κB) and IκBα were detected by immunofluorescence.ResultsThe FFA mixture (oleate/palmitate, at a 2:1 ratio) of 0.5 mmol/L had the highest cell viability and fat accumulation rate, which was preferable for establishing an in vitro fatty liver model. The expression of inflammatory factors (TNF-α and IL-6) and oxidants Malonaldehyde (MDA), 4-hydroxynonenal (HNE) and reactive oxygen species (ROS) also increased in the L-02 fatty liver cells. The expression levels of aging markers and aging genes, such as SA-β-gal, p16, p21, p53 and pRb, increased more in the L-02 fatty liver cells than in the L-02 cells. The total levels of the apoptosis-associated proteins Bcl2, Bax, Bax/Bcl-2, CyCt and cleaved caspase 9 were also upregulated in the L-02 fatty liver cells. All of the above genes and proteins were downregulated in the huperzine A and FFA co-treatment group. In the L-02 fatty liver cells, the expression of IκBα decreased, while the expression of NF-κB increased. After the huperzine A and FFA co-treatment, the expression of IκBα increased, while the expression of NF-κB decreased.ConclusionFatty liver cells showed an obvious senescence and apoptosis phenomenon. Huperzine A suppressed hepatocyte senescence, and it might exert its anti-aging effect via the NF-κB pathway.

[Background and aims]NAFLD is a common chronic liver disease. The purpose of this study is to carry out a current large-scale epidemiological survey and analyse factors related to the incidence and the severity of NAFLD.[Methods]Three communities of Shanghai were selected by stratified random sampling in 2019.The residents older than 18 years old underwent medical history taking, physical examination, laboratory examination and liver ultrasonography examination. The prevalence of different degrees of NAFLD, the prevalence of metabolic factors, and factors associated with the severity of NAFLD was analysed. [Results]A total of 19250 subjects were enrolled. The total prevalence of NAFLD in Shanghai was 45.55%, 62.00% in males and 27.54% in females. Age, FLI, BMI, WC, ALT, AST, AKP, GGT, CR, FPG, UA, TC, TG and LDL in NAFLD patients were higher than those in non-NAFLD people. The distribution of NAFLD prevalence was different in different age groups. The prevalence of diabetes, obesity and hypertriglyceridemia in the NAFLD group were higher than that in the non-NAFLD group. In males, FLI, weight, WC, ALT, UA, TG, GGT and metabolic syndrome were positively correlated with the severity of NAFLD; while HDL was negatively correlated with it. In females, age, FLI, weight, WC, ALT, FPG, UA, TG, LDL, GGT, AKP and metabolic syndrome were positively correlated with the severity of NAFLD; while HDL was negatively correlated with it.[Conclusions]The prevalence of NAFLD in Shanghai is high. The distribution of NAFLD prevalence was different in different age groups. Many factors are related to the incidence of NAFLD and the severity of NAFLD.