Asunto(s)
Autofagia/efectos de los fármacos , Berberina/uso terapéutico , Estrés del Retículo Endoplásmico/efectos de los fármacos , Hígado Graso/tratamiento farmacológico , Trasplante de Hígado , Animales , Autofagosomas/ultraestructura , Retículo Endoplásmico/efectos de los fármacos , Retículo Endoplásmico/ultraestructura , Hígado/efectos de los fármacos , Hígado/patología , Masculino , Microscopía Electrónica de Transmisión , Microscopía Fluorescente , Estrés Oxidativo/efectos de los fármacos , Ratas , Ratas Wistar , Tapsigargina/farmacologíaRESUMEN
Berberine (BBR) has been demonstrated to protect against renal ischemia/reperfusion injury; however, the underlying molecular mechanism is largely unknown. In the present study, we examined the role of silent information regulator 1 (Sirt1)/p53 in the protective effect of BBR on hypoxia/reoxygenation (H/R)-mediated mitochondrial dysfunction in rat renal tubular epithelial cells (NRK-52E cells). NRK-52E cells were preconditioned with small interfering RNA targeting Sirt1 (Sirt1-siRNA) and BBR before subjected to H/R. Cell damage was assessed by CCK8 assay and detection of oxidative parameters. The apoptotic rate was determined by flow cytometry and Hoechst 33258 staining. The expression of apoptotic markers, Sirt1, p53 and the translocation of p53 were examined by Western blotting assay. Nuclear p53 deacetylation by Sirt1 was detected using immunoprecipitation. Compared with the H/R group, BBR pretreatment increased cell viability and inhibited mitochondrial oxidative stress and apoptosis. Protein expression of Sirt1 was also enhanced along with a reduction of p53. Furthermore, both nuclear translocation of p53 and its acetylation were inhibited in NRK-52E cells pretreated with BBR. However, the knockdown of Sirt1 counteracted the renoprotection of BBR. BBR preconditioning protects rat renal tubular epithelial cells against H/R-induced mitochondrial dysfunction via regulating the Sirt1/p53 pathway.
Asunto(s)
Berberina/uso terapéutico , Hipoxia de la Célula/efectos de los fármacos , Sustancias Protectoras/uso terapéutico , Daño por Reperfusión/metabolismo , Sirtuina 1/metabolismo , Proteína p53 Supresora de Tumor/metabolismo , Apoptosis , Berberina/farmacología , Humanos , Sustancias Protectoras/farmacología , TransfecciónRESUMEN
The inducible coactivator PGC-1 α plays master regulator in mitochondrial biogenesis and thermogenesis in brown adipose tissues (BATs). BAT is a natural antiobesity organ which dissipates chemical energy in the form of heat through specialized mitochondrial protein UCP-1. Eletroacupuncture (EA) has been widely used as an alternative treatment for obesity and its related disorders such as type 2 diabetes. The molecular mechanism of electroacupuncture on treatment of obesity is still unclear. We hypothesized that electroacupuncture induced PGC-1 α expression to increase the energy expenditure in BAT. Rats were randomly divided into control group and electroacupuncture treatment group. We investigated the effects of electroacupuncture at Zusanli (ST36) acupoint on the expressions of PGC-1 α and its associated genes in the BAT of rats using real-time PCR and western blotting. We found that electroacupuncture effectively induces the expression of PGC-1 α and UCP-1 by 4-fold and 5-fold in the BAT of rats, respectively. Our results indicated that the molecular mechanism of electroacupuncture for the treatment of obesity may be, or at least partially, through induction of both PGC-1 α and UCP-1 expressions to increase energy expenditure in BAT.