Objective： To investigate the neuroprotective effect of Bu-Shen-Huo-Xue （BSHX） extract, a polyherbal formula, against High Glucose （HG）-induced neurotoxicity in PC12 cells. Methods： Cell viability assay, Lactate...Objective： To investigate the neuroprotective effect of Bu-Shen-Huo-Xue （BSHX） extract, a polyherbal formula, against High Glucose （HG）-induced neurotoxicity in PC12 cells. Methods： Cell viability assay, Lactate Dehydrogenase （LDH） assay, Reactive Oxygen Species （ROS） detection, Hoechst 33258, Acridine Orange （AO）/Ethidium Bromide （EB） double stain and Mitochondrial Membrane Potential （MMP） assay were performed. In addition, Bax, Bcl-2, caspase-3, cleaved caspase-3, PARP, cleaved PARP, cytochrome c and Mitogen-Activated Protein Kinases （MAPKs） were detected by western blot. Results： BSHX extract increased cell viability and decreased LDH leakage in a concentration-dependent manner in HG-induced PC12 cells. Moreover, BSHX extract decreased the level of intracellular ROS, increased mitochondrial membrane potential, regulated the expressions of Bax and Bcl-2, and inhibited the release of cytochrome c from mitochondria. Furthermore, BSHX extract attenuated the activation of caspase-3 and PARP, and inhibited the phosphorylations of c-Jun N-terminal kinase （JNK） and p38 MAPKs. Conclusion： BSHX extract exhibited significant neuroprotective effect on HG-induced apoptosis in PC12 cells. This effect may be associated with the suppression of ROS generation as well as mitochondria-mediated caspase and JNK/p38 MAPK signaling pathways.展开更多
基金This work was supported by the National Natural Science Foundation of China （Grant Nos. 81530099, 81573763 and 81773932）, the Beijing Municipal Natural Science Foundation （Grant No.7172221）, and the National Key Technology R＆D Program ＂New Drug Innovation＂ of China （Grant No. 2016YFE0116200）.
文摘Objective： To investigate the neuroprotective effect of Bu-Shen-Huo-Xue （BSHX） extract, a polyherbal formula, against High Glucose （HG）-induced neurotoxicity in PC12 cells. Methods： Cell viability assay, Lactate Dehydrogenase （LDH） assay, Reactive Oxygen Species （ROS） detection, Hoechst 33258, Acridine Orange （AO）/Ethidium Bromide （EB） double stain and Mitochondrial Membrane Potential （MMP） assay were performed. In addition, Bax, Bcl-2, caspase-3, cleaved caspase-3, PARP, cleaved PARP, cytochrome c and Mitogen-Activated Protein Kinases （MAPKs） were detected by western blot. Results： BSHX extract increased cell viability and decreased LDH leakage in a concentration-dependent manner in HG-induced PC12 cells. Moreover, BSHX extract decreased the level of intracellular ROS, increased mitochondrial membrane potential, regulated the expressions of Bax and Bcl-2, and inhibited the release of cytochrome c from mitochondria. Furthermore, BSHX extract attenuated the activation of caspase-3 and PARP, and inhibited the phosphorylations of c-Jun N-terminal kinase （JNK） and p38 MAPKs. Conclusion： BSHX extract exhibited significant neuroprotective effect on HG-induced apoptosis in PC12 cells. This effect may be associated with the suppression of ROS generation as well as mitochondria-mediated caspase and JNK/p38 MAPK signaling pathways.
