RESUMEN
Oxidative stress can cause the excessive generation of reactive oxygen species (ROS) and has various adverse effects on muscular mitochondria. Qiangji Jianli decoction (QJJLD) is an effective traditional Chinese medicine (TCM) that is widely applied to improve muscle weakness, and it has active constituents that prevent mitochondrial dysfunction. To investigate the protective mechanism of QJJLD against hydrogen peroxide- (H2O2-) mediated mitochondrial dysfunction in L6 myoblasts. Cell viability was determined with MTT assay. Mitochondrial ultrastructure was detected by transmission electron microscope (TEM). ROS and mitochondrial membrane potential (MMP) were analyzed by fluorescence microscope and flow cytometry. The superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) activity, and malondialdehyde (MDA) level were determined by WST-1, TBA, and DTNB methods, respectively. The mRNA and protein levels were measured by quantitative real-time PCR (qRT-PCR) and Western blot. The cell viability was decreased, and the cellular ROS level was increased when L6 myoblasts were exposed to H2O2. After treatment with QJJLD-containing serum, the SOD and GSH-Px activities were increased. MDA level was decreased concurrently. ROS level was decreased while respiratory chain complex activity and ATP content were increased in L6 myoblasts. MMP loss was attenuated. Mitochondrial ultrastructure was also improved. Simultaneously, the protein expressions of p-AMPK, PGC-1α, NRF1, and TFAM were upregulated. The mRNA and protein expressions of Mfn1/2 and Opa1 were also upregulated while Drp1 and Fis1 were downregulated. These results suggest that QJJLD may alleviate mitochondrial dysfunction through the regulation of mitochondrial dynamics and biogenesis, the inhibition of ROS generation, and the promotion of mitochondrial energy metabolism.
Asunto(s)
Antígenos de Superficie/metabolismo , ADN Mitocondrial/efectos adversos , Medicamentos Herbarios Chinos/uso terapéutico , Peróxido de Hidrógeno/efectos adversos , Proteínas de Neoplasias/metabolismo , Animales , Medicamentos Herbarios Chinos/farmacología , Humanos , Dinámicas Mitocondriales/efectos de los fármacos , Mioblastos/metabolismo , Biogénesis de Organelos , RatasRESUMEN
The Qiangji Jianli Decoction (QJJLD) is an effective Chinese medicine formula for treating Myasthenia gravis (MG) in the clinic. QJJLD has been proven to regulate mitochondrial fusion and fission of skeletal muscle in myasthenia gravis. In this study, we investigated whether QJJLD plays a therapeutic role in regulating mitochondrial biogenesis in MG and explored the underlying mechanism. Rats were experimentally induced to establish autoimmune myasthenia gravis (EAMG) by subcutaneous immunization with R97-116 peptides. The treatment groups were administered three different dosages of QJJLD respectively. After the intervention of QJJLD, the pathological changes of gastrocnemius muscle in MG rats were significantly improved; SOD, GSH-Px, Na+-K+ ATPase and Ca2+-Mg2+ ATPase activities were increased; and MDA content was decreased in the gastrocnemius muscle. Moreover, AMPK, p38MAPK, PGC-1α, NRF-1, Tfam and COX IV mRNA and protein expression levels were also reversed by QJJLD. These results implied that QJJLD may provide a potential therapeutic strategy through promoting mitochondrial biogenesis to alleviate MG via activating the AMPK/PGC-1α signaling pathway.
Asunto(s)
Proteínas Quinasas Activadas por AMP/metabolismo , Medicamentos Herbarios Chinos/farmacología , Mitocondrias Musculares/efectos de los fármacos , Músculo Esquelético/efectos de los fármacos , Miastenia Gravis Autoinmune Experimental/tratamiento farmacológico , Biogénesis de Organelos , Coactivador 1-alfa del Receptor Activado por Proliferadores de Peroxisomas gamma/metabolismo , Proteínas Quinasas Activadas por AMP/genética , Animales , Femenino , Regulación de la Expresión Génica , Mitocondrias Musculares/enzimología , Mitocondrias Musculares/genética , Mitocondrias Musculares/ultraestructura , Músculo Esquelético/enzimología , Músculo Esquelético/ultraestructura , Miastenia Gravis Autoinmune Experimental/enzimología , Miastenia Gravis Autoinmune Experimental/inmunología , Miastenia Gravis Autoinmune Experimental/patología , Fragmentos de Péptidos , Coactivador 1-alfa del Receptor Activado por Proliferadores de Peroxisomas gamma/genética , Ratas Endogámicas Lew , Receptores Colinérgicos , Transducción de SeñalRESUMEN
Blueberry pomace is abundant in anthocyanins. This work characterized the anthocyanins in blueberry pomace, discussed the stability of anthocyanins under ultrasound treatment, and compared the extraction conditions for different anthocyanin compositions. Thirteen anthocyanins were identified, and malvidin-3-galactoside (18.56%), which represented the most abundant anthocyanin, was selected as the individual analyte. The general linear model univariate analysis revealed that ultrasound-assisted extraction (UAE) resulted in higher recoveries of both total anthocyanins (TA) and individual anthocyanins (IA) when compared with conventional solvent extraction. The optimized extraction conditions for TA and IA were UAE in pure methanol (12.49 mg/g dry weight) at 25 °C for 30 min and UAE in 70% ethanol (3.57 mg/g dry weight) at 40 °C for 40 min, respectively. Moreover, IA was more vulnerable to degradation compared with TA. Therefore, a specific extraction process of IA is significant for monomer preparation, and harsh conditions should be avoided in UAE.