Protective Effects Of Ginsenoside Rg1 On Astrocytes And Rg1 Pretreated Cerebral Ischemic-reperfusion Mice

ObjectiveStudy the protective effect of Rg1 on astrocytes and Rg1 pretreated cerebral ischemia-reperfusion mice to explore the related mechanisms preliminarily.Methods1. Effects of Rg1 on astrocytesIn this study, fluorescence enzyme-labelled meter plus Sytox Green were used to measure the effects of Rg1 on apoptosis of astrocytes induced by CHX, CTX and H2O2. With xCELLigence RTCA System, we investigated the protective effects of Rg1 on astrocytes which were stimulated by Ionomycin and H2O2. We also detected the influence of Rg1 on Ca2+ entry to astrocytes in resting condition by Flow Cytometry. Using Confocal Laser Scanning Microscope (LSM), we detected the inhibition of Rgl on Ca2+ overload, and then examined the reductive effect of Rg1 on LPS-induced ROS release by Flow Cytometry. Using Flow Cytometry and LSM, we detected the influence of Rg1 on mitochondrial membrane potential and Ca2+ overload in mitochondrial of astrocytes induced by Ion.2. Effects of Rg1 on Rg1 pretreated cerebral ischemia-reperfusion miceBrain infarct areas of Rg1 pretreated cerebral ischemia-reperfusion mice were detected by TTC staining. With dry/wet specific gravity method, we tested brain water content (bwc) of Rg1 pretreated cerebral ischemia-reperfusion mice. Cerebral ischemia-reperfusion mice preinjected with Rg1 and unpreinjected with Rg1 were both bred under the same condition and the life-span was calculated.Results1. Protrctive effects of Rg1 on astrocytesRg1 didn't have any cell demage effects on cultured astrocytes in the concentration of 10μmol/L. Rg1 could inhibit the apoptosis of astrocytes induced by CHX, CTX and H2O2. Rg1 inhibited Ca2+ overload in cultured astrocytes after stimulation and enhanced cell survive ability at the same time, which was detected by Confocol Laser Scanning Microscrobe (LSM) and Flow Cytometry (FACS). Rg1 could trigger a low-level Ca2+ influx in resting condition detected by FACS. Rg1 could inhibit ROS release detected by FACS. Detected by FACS and LSM, we also detected that Rgl inhibited the reduction of mitochondrial membrane potential and Ca2+ overload in mitochondrial induced by Ion.2. Protective effects of Rgl on Rg1 pretreated cerebral ischemia-reperfusion miceThe infarct size and brain edema of Rg1 pretreated cerebral ischemia-reperfusion mice were obviously smaller than those unpretreated with Rg1. Rg1 also prolonged life time of Rg1 preinjected cerebral ischemia-reperfusion mice.