XinJiaCongRongTuSiZiWan protects triptolide-induced rats from oxidative stress injury via mitophagy mediated PINK1/Parkin signaling pathway.

PURPOSE: XinJiaCongRongTuSiZiWan (XJCRTSZW) is a traditional Chinese medicine compound for invigorating the kidney, nourishing blood, and promoting blood circulation. This study aimed to explore the effect of XJCRTSZW on triptolide (TP)-induced oxidative stress injury. METHODS: Adult female Sprague-Dawley rats and human ovarian granulosa cell lines were treated with TP and XJCRTSZW. Hematoxylin and eosin staining, enzyme-linked immunosorbent assay, flow cytometry, CCK-8, JC-1 staining, transmission electron microscopy, reverse transcription-quantitative polymerase chain reaction, and Western blotting were performed in this study. RESULTS: XJCRTSZW treatment observably ameliorated the TP-induced pathological symptoms. Furthermore, XJCRTSZW treatment observably enhanced the TP-induced reduction of estradiol, anti-Mullerian hormone, progesterone, superoxide dismutase, ATP content, mitochondrial membrane potential, p62, and Hsp60 mRNA, and protein levels in vivo and in vitro (p < 0.05). However, TP-induced elevation of follicle stimulating hormone and luteinizing hormone concentrations, malondialdehyde levels, reactive oxygen species levels, apoptosis rate, mitophagy, and the mRNA and protein expressions of LC3-II/LC3-I, PTEN-induced kinase 1 (PINK1), and Parkin were decreased (p < 0.05). In addition, XJCRTSZW treatment markedly increased cell viability in vitro (p < 0.05). CONCLUSIONS: XJCRTSZW protects TP-induced rats from oxidative stress injury via the mitophagy-mediated PINK1/Parkin pathway.

Regulation of Ca2+ uptake in skeletal muscle by 1,25-dihydroxyvitamin D3: role of phosphorylation and calmodulin.

Experiments were carried out to obtain information about the mechanism underlying the fast action of 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) in skeletal muscle. N-2'-o-dibutyryladenosine-3',5'-cyclic monophosphate (dbcAMP), similarly as 1,25(OH)2D3 (5 x 10(-10) M), rapidly increased 45Ca uptake by soleus muscle from vitamin D-deficient chicks (+25% and +98% at 3 min and 10 min, respectively) in a dose-dependent manner. The effects of the cAMP analog (10 microM) and 1,25(OH)2D3 could be abolished by the Ca(2+)-channel blocker nifedipine and the calmodulin antagonist flufenazine. Calmodulin binding by two muscle microsomal proteins of 28 kDa and 30 kDa was stimulated within 1 min of exposure of the tissue to 1,25(OH)2D3. Direct effects of the sterol on membrane calmodulin binding were shown with isolated microsomes. The 1,25(OH)2D3-mediated rise of [125I]calmodulin binding to microsomal membranes was dependent on the presence of medium ATP. Forskolin (10 microM) and cAMP (10 microM) also increased [125I]calmodulin binding (+75% and +64%, respectively, with respect to controls). Pretreatment of microsomal membranes with cAMP-dependent protein kinase inhibitor (1 microgram/ml) or addition of alkaline phosphates (1 U/ml) after hormonal treatment caused complete inhibition of 1,25(OH)2D3-induced [125I]calmodulin binding to microsomal membrane proteins. These results imply modifications of membrane protein phosphorylation through the cAMP signal pathway and in turn of calmodulin binding in the mechanism by which 1,25(OH)2D3 rapidly stimulates skeletal muscle Ca2+ uptake.

Insulin and IGF-I stimulated RNA synthesis in primary cultures of neuronal cells: involvement of cyclic AMP and protein kinase-C.

Insulin and insulin-like growth factor I promote the growth of rat neuronal cells in primary culture. In order to investigate the mechanism of hormone signalling in this biological system, we studied the effect of cyclic AMP agonists and a protein kinase C stimulator on basal and hormone-induced RNA synthesis. Agents elevating endogenous cyclic AMP levels (forskolin, dibutyryl cyclic AMP, cholera toxin) blocked the stimulatory effects of both insulin and the growth factor; dibutyryl cyclic AMP, however, altered the binding of neither hormone. Although, unlike the aforementioned agents, phorbol ester significantly elevated basal RNA synthesis; it nevertheless inhibited the stimulation by insulin; this latter effect probably being mediated by an increase in intracellular cyclic AMP levels, as has been found in other cell types. Staurosporine, an inhibitor of protein kinase C, also blocked the effects of insulin on RNA synthesis.

Insulin and IGF-I stimulated RNA synthesis in primary cultures of neuronal cells: involvement of cyclic AMP and protein kinase-c

La insulina y el IGF-I promueven el crecimiento de las células neuronales de rata en cultivo primario. Con el objeto de investigar el mecanismo de transducción de señales hormonales en este sistema biológico, estudiamos el efecto de agonistas de AMP cíclico y un estimulador de la proteína kinasa-C sobre la síntesis de ARN basal e inducida por hormonas. Los agentes que aumentan los níveles de AMP cíclico endógenos (foraskolina, dibutiril-AMP cíclico, toxina colérica) bloquearon los efectos estimuladores de la insulina y el factor de crecimiento; el dibutiril AMP cíclico, sin embargo, no alteró la unión de las hormonas a sus receptores. Aunque a diferencia de los agentes antes mencionados, el ester de forbol elevó significativamente la síntesis de ARN basal; este, no obstante, inhibió la estimulación por la insulina. Este último efecto probablemente fue mediado por un incremento en los niveles de AMP cíclico, como se ha encontrado en otros tipos de células. La estaurosporina, un inhibidor de la proteína kinasa-C, también bloqueó los efectos de la insulina sobre la síntesis de RNA

Insulin and IGF-I stimulated RNA synthesis in primary cultures of neuronal cells: involvement of cyclic AMP and protein kinase-c

La insulina y el IGF-I promueven el crecimiento de las células neuronales de rata en cultivo primario. Con el objeto de investigar el mecanismo de transducción de señales hormonales en este sistema biológico, estudiamos el efecto de agonistas de AMP cíclico y un estimulador de la proteína kinasa-C sobre la síntesis de ARN basal e inducida por hormonas. Los agentes que aumentan los níveles de AMP cíclico endógenos (foraskolina, dibutiril-AMP cíclico, toxina colérica) bloquearon los efectos estimuladores de la insulina y el factor de crecimiento; el dibutiril AMP cíclico, sin embargo, no alteró la unión de las hormonas a sus receptores. Aunque a diferencia de los agentes antes mencionados, el ester de forbol elevó significativamente la síntesis de ARN basal; este, no obstante, inhibió la estimulación por la insulina. Este último efecto probablemente fue mediado por un incremento en los niveles de AMP cíclico, como se ha encontrado en otros tipos de células. La estaurosporina, un inhibidor de la proteína kinasa-C, también bloqueó los efectos de la insulina sobre la síntesis de RNA (AU)