Background Cerebral ischemia-reperfusion injury is the main reason for the loss of neurons in the ischemic cerebrovascular disease. Therefore, to deeply understand its pathogenesis and find a new target is the key iss...Background Cerebral ischemia-reperfusion injury is the main reason for the loss of neurons in the ischemic cerebrovascular disease. Therefore, to deeply understand its pathogenesis and find a new target is the key issue to be solved. This research aimed to investigate the neuroprotective effects of salvianolic acid B （SalB） against oxygen-glucose deprivation/reperfusion （OGD/RP） damage in primary rat cortical neurons.Methods The primary cultures of neonatal Wister rats were randomly divided into the control group, the OGD/RP group and the SalB-treatment group （10 mg/L）. The cell model was established by depriving of oxygen and glucose for 3 hours and reperfusion for 3 hours and 24 hours, respectively. The neuron viability was determined by MTT assay. The level of cellular reactive oxygen species （ROS） was detected by fluorescent labeling method and spin trapping technique respectively. The activities of neuronal Mn-superoxide dismutase （Mn-SOD）, catalase （CAT） and glutathione peroxidase （GSH-PX） were assayed by chromatometry. The mitochondria membrane potential （△ψm） was quantitatively analyzed by flow cytometry. The release rate of cytochrome c was detected by Western blotting. The neuronal ultrastructure was observed by transmission electron microscopy. Statistical significance was evaluated by analysis of variance （ANOVA）followed by Student-Newman-Keuls test.Results OGD/RP increased the level of cellular ROS, but decreased the cell viability and the activities of Mn-SOD, CAT and GSH-PX; SalB treatment significantly reduced the level of ROS （P ＜0.05）; and enhanced the cell viability （P ＜0.05）and the activities of these antioxidases （P ＜0.05）. Additionally, OGD/RP induced the fluorescence value of △ψm to diminish and the release rate of cytochrome c to rise notably; SalB markedly elevated the level of △ψm （P ＜0.01） and depressed the release rate of cytochrome c （P ＜0.05）; it also ameliorated the neuronal morphological injury.Conclusion The neuroprotection of SalB may be attributed to the elimination of ROS and the inhibition of apoptosis.展开更多
Background The elevated matrix metalloproteinase （MMP） activity is an important cause of chronic wound healing failure. Arsenolite, whose main component is arsenic trioxide （As203）, is a common traditional Chinese...Background The elevated matrix metalloproteinase （MMP） activity is an important cause of chronic wound healing failure. Arsenolite, whose main component is arsenic trioxide （As203）, is a common traditional Chinese medicine wildly used in treating chronic wounds; it can remove necrotic tissue and promote tissue regeneration. This research was designed to evaluate the effects of AS2O3 on production and activities of MMP-1, MMP-2 and MMP-9, and on regulation of its signal transduction pathway in human skin fibroblasts （HSFb） and human monocyte line （THP-1 cells） that were in an inflammatory state. Methods We established three cell models; HSFb activated by TNF-α, THP-1 cells activated by phorbol 12-myristate 13-acetate （PMA） and an HSFb-THP-1 co-culture system. Three cell models was cultured with AS203 for 24 hours. The levels of MMP-1, MMP-2, MMP-9, TNF-α and IL-1β in the cell culture supernatants were assayed by ELJSA. The mRNA expressions of MMP-1, MMP-2 and MMP-9 were determined by RT-PCR. The activities of MMP-2 and MMP-9 were tested by Gelatin zymography assays. The phosphorylation levels of ERK1/2 and p38MAPK were assayed by Western blotting. Results As2O3 inhibited the expression of MMP-1, MMP-2 and MMP-9 mRNA, the secretion and activity of MMP-1, MMP-2 and MMP-9 in HSFb and THP-1 cells in the inflammatory state （P 〈0.05 and P 〈0.01 respectively）. It also inhibited the secretion of TNF-α and IL-1β in THP-1 cells and in the co-culture system （P 〈0.05 and P 〈0.01, respectively）. It also decreased the phosphorylation of ERK1/2 and p38 MAPK in HSFb and THP-1 cells （P 〈0.05 and P 〈0.01, respectively）. Conclusions AS203, as a main component of arsenolite, can inhibit the production of MMPs by HSFb and THP-1 cells in an inflammatory state through inhibiting the release of inflammatory factors and the activation of the MAPK cascade pathway. This may be a possible mechanism for arsenolite healing chronic wounds.展开更多
基金This work was supported by a grant from the National Natural Science Foundation of China （No. 30472281）.
文摘Background Cerebral ischemia-reperfusion injury is the main reason for the loss of neurons in the ischemic cerebrovascular disease. Therefore, to deeply understand its pathogenesis and find a new target is the key issue to be solved. This research aimed to investigate the neuroprotective effects of salvianolic acid B （SalB） against oxygen-glucose deprivation/reperfusion （OGD/RP） damage in primary rat cortical neurons.Methods The primary cultures of neonatal Wister rats were randomly divided into the control group, the OGD/RP group and the SalB-treatment group （10 mg/L）. The cell model was established by depriving of oxygen and glucose for 3 hours and reperfusion for 3 hours and 24 hours, respectively. The neuron viability was determined by MTT assay. The level of cellular reactive oxygen species （ROS） was detected by fluorescent labeling method and spin trapping technique respectively. The activities of neuronal Mn-superoxide dismutase （Mn-SOD）, catalase （CAT） and glutathione peroxidase （GSH-PX） were assayed by chromatometry. The mitochondria membrane potential （△ψm） was quantitatively analyzed by flow cytometry. The release rate of cytochrome c was detected by Western blotting. The neuronal ultrastructure was observed by transmission electron microscopy. Statistical significance was evaluated by analysis of variance （ANOVA）followed by Student-Newman-Keuls test.Results OGD/RP increased the level of cellular ROS, but decreased the cell viability and the activities of Mn-SOD, CAT and GSH-PX; SalB treatment significantly reduced the level of ROS （P ＜0.05）; and enhanced the cell viability （P ＜0.05）and the activities of these antioxidases （P ＜0.05）. Additionally, OGD/RP induced the fluorescence value of △ψm to diminish and the release rate of cytochrome c to rise notably; SalB markedly elevated the level of △ψm （P ＜0.01） and depressed the release rate of cytochrome c （P ＜0.05）; it also ameliorated the neuronal morphological injury.Conclusion The neuroprotection of SalB may be attributed to the elimination of ROS and the inhibition of apoptosis.
基金This study was supported by a grant from the Beijing Natural Science Foundation （No. 7072014）.
文摘Background The elevated matrix metalloproteinase （MMP） activity is an important cause of chronic wound healing failure. Arsenolite, whose main component is arsenic trioxide （As203）, is a common traditional Chinese medicine wildly used in treating chronic wounds; it can remove necrotic tissue and promote tissue regeneration. This research was designed to evaluate the effects of AS2O3 on production and activities of MMP-1, MMP-2 and MMP-9, and on regulation of its signal transduction pathway in human skin fibroblasts （HSFb） and human monocyte line （THP-1 cells） that were in an inflammatory state. Methods We established three cell models; HSFb activated by TNF-α, THP-1 cells activated by phorbol 12-myristate 13-acetate （PMA） and an HSFb-THP-1 co-culture system. Three cell models was cultured with AS203 for 24 hours. The levels of MMP-1, MMP-2, MMP-9, TNF-α and IL-1β in the cell culture supernatants were assayed by ELJSA. The mRNA expressions of MMP-1, MMP-2 and MMP-9 were determined by RT-PCR. The activities of MMP-2 and MMP-9 were tested by Gelatin zymography assays. The phosphorylation levels of ERK1/2 and p38MAPK were assayed by Western blotting. Results As2O3 inhibited the expression of MMP-1, MMP-2 and MMP-9 mRNA, the secretion and activity of MMP-1, MMP-2 and MMP-9 in HSFb and THP-1 cells in the inflammatory state （P 〈0.05 and P 〈0.01 respectively）. It also inhibited the secretion of TNF-α and IL-1β in THP-1 cells and in the co-culture system （P 〈0.05 and P 〈0.01, respectively）. It also decreased the phosphorylation of ERK1/2 and p38 MAPK in HSFb and THP-1 cells （P 〈0.05 and P 〈0.01, respectively）. Conclusions AS203, as a main component of arsenolite, can inhibit the production of MMPs by HSFb and THP-1 cells in an inflammatory state through inhibiting the release of inflammatory factors and the activation of the MAPK cascade pathway. This may be a possible mechanism for arsenolite healing chronic wounds.
