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Aim To study the inhibitory effect of ginsenoside Rgl on neuronal ferroptosis after ischemic stroke and its mechanism. Methods A model of oxygen glucose deprivation/reoxygenation (OGD/R) was established in HT22 cells, and the effect of Rgl on the viability of HT22 cells after OGD/R injury was detected by CCK-8. The effect of Rgl on ferroptosis in HT22 cells after OGD/R injury was detected by the test of ferroptosis markers GSH/GSSG, SOD, MDA, and Fe
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Aim To investigate the mechanism by which ginsenoside Rgl regulates autophagy anrl delays brain aging in mice through AMPK/mTOR signaling pathway.Methods C57BL/6J male mice were ran¬domly divided into four groups, namely brain aging model group ,control group, Rgl anti-aging group,auto¬phagy activator Rapamycin anti-aging group.After the modeling was completed, the test of each experimental index would be carried out on the next day.Morris wa¬ter maze experiment was used to detect the learning and memory ability of mice.Paraffin sections of the hippocampus were prepared, HE , Nissl and immunohis- tochemical staining were used to observe the morpholo¬gy of hippocampal neurons, the number of neurons and Nissl bodies was counted, and autophagy-related proteins p62 , ATG5 , ULK1 were detected.Brain tissue homogenates were prepared to detect the aetivity of brain tissue acetylcholinesterase ( AChE ).Western blot was userl to detect brain tissue autophagy-related proteins LC3II, P62, beclinl, P-AMPK/AMPK, P- mTOR/mTOR and apoptosis protein P53.Results Water maze test showed that Rgl and Hap significantly improved the learning and memory abilities of brain-ag¬ing mice.HE and Nissl staining showed that Rgl and Rap decreased necrotic cells and increased the number of Nissl bodies in the hippocampus of brain-aging mice.Immunohistochemistry staining showed that Rgl and Rap decreased the expression of neuronal autoph- agv protein P62 in hippocampus and increased the ex-pression of ATG5 and ULK1.Rgl and Rap decreased the activity of AhcE in brain-aging mice.Western blot showed that Rgl and Rap increased autophagy-related proteins LC3II, Beclinl , P-AMPK/AMPK, but de¬creased the expression of P-mTOR/mTOR, P62, P53.Conclusions Ginsenoside Rgl can effectively antago¬nize the aging effect of D-gal on mouse brain.The pos¬sible mechanism is related to the regulation of autoph- agv by Rgl through AMPK/mTOR signaling pathway.
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Aim To investigate the effect of ginsenoside Rgl on PC 12 cell hypoxia-reoxygenation injury and its possible mechanism. Methods PC 12 cells were randomly divided into six groups. Except for the blank control group, all the other groups were hypoxia and hypoglycemia for 6 hours, and then reoxygenated and glycosylated for 24 hours to make OGD/R models. Each drug group was given corresponding drugs 2 hours before modeling pretreatment. DCFH-DA method was used to detect the ROS production in cells, Annexin V- FITC/PI double staining method was performed to detect cell apoptosis rate, ELISA method was used to detect LDH activity and IL-1 (3 content in cell supernatant, and Western blot was applied to detect the ex- pression of proteins of N0X2, p22phox, p47phox, NLRPl, ASC, Caspase-1, PSD95, Tau, p-Tau and observe the intervention effect of ginsenoside Rgl. Re sults Tempol, Apocynin and Rgl (5, 10 jjLmol • L"1) groups could significantly inhibit ROS production and apoptosis, reduce LDH release and IL-1 (3 content in cell supernatant; Apocynin and Rgl (5, 10 |xmol • L"1) groups could significantly down-regulate the expression of N0X2, p22phox and p47phox in cells. The Tempol, Apocynin and Rgl (5, 10 jxmol • L"1 ) groups could significantly decrease the protein expression of NLRP1, Caspase-1, ASC, IL-1 (3 and p-Tau, and markedly down-regulate the expression of PSD95 protein. Conclusion Rgl is likely to reduce the is- chemia-reperfusion injury of PC 12 cells by inhibiting the NOX2-NLRP1 pathway.
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To establish a model of ocular apoptosis in zebrafish by using dibutyl phthalate(DBP), and accordingly to evaluate the activity of anti-ocular apoptosis drugs. Methods Zebrafish embryos (30 hpf) were treated with DBP(0. 1, 10, 50, 100 jxmol L " 1) for 18 h and 42 h, and the survival rate was calculated. Then zebrafish embryos (30 or 54 hpf) were incubated with DBP(0. 1, 1 , 5 , 10, 50 |j.mol L 1) for 18 h under direct or indirect light conditions, and visualized under microscope, then the ocular apoptosis was assessed by AO staining. Furthermore, modeled zebrafish was treated with N-acetyl-L-cysteine(NAC) or ginsenoside Rgl, and the effects on ocular apoptosis were observed. Results There was no mortality in zebrafish at concentrations below 10 |j,mol L"1 after modeling with DBP for 18 h and 42 h. The survival rate of zebrafish was significantly reduced with a concentration above 50 |xmol L"1 (P < 0 . 0 1) . Furthermore, obvious ocular apoptosis was found at 30 hpf after 18 h with DBP(10 (unol L"1) (P < 0. 0 1) . The zebrafish showed spontaneous apoptosis of the ocular cells at 72 h, which was not associated with light conditions. Effect of NAC or ginsenoside Rgl was observed on decreasing ocular apoptosis in zebrafish (P < 0. 05) . Conclusions DBP can be used to rapidly and efficiently establish a model of ocular apoptosis in zebrafish, which can be used for rapid evaluation of drug activity-.
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Objective To investigate the influence of ginsenoside Rgl on the proliferation of neural stem cells in adult rats with fo- cal cerebral ischemia,and to explore the possible mechanism of ginsenoside Rg1 in brain protection and anti-aging action.Methods The animal model of left middle cerebral artery occlusion(MCAO)was reproduced in adult male Wistar rats.The thymidine analog bromode- oxyuridine(BrdU)was administered intraperitoneally(50mg/kg,Sigma,USA)every four hours for four times before executing the ani- mals to label the proliferating cells.By the employment of immunohistochemical single staining and double-immunofluorescence technique, the number of BrdU immunoreacted nuclei in the subventricular zone(SVZ),the number of nestin positive cells in the subgranular zone (SGZ)as well as the nestin/BrdU double-labeled positive cells were counted.The effect of ginsenoside Rg1 on the immunoreactivity for BrdU,nestin and nestin/BrdU at 1d,3d,7d and 14d after focal cerebral ischemia were also observed to compare with that of controls,Re- sults The immunoreacted cells of BrdU,nestin and nestin/BrdU were seen in SVZ and SGZ in the hippocampus following focal cerebral ischemia.The number of BrdU,nestin-positive cells and the double-labeled positive cells all reached the peak on 7d and decreased on 14d after MCAO,and the expressions markedly increased after the rats were given ginsenoside Rg1,compared with the control(P
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Ginseng has long been used as a tonic and agent for prolonging life span in chinese traditional medicine． Using morden technology，ginsenoside Rgl and Rbl were proved to be main active principles of ginseng．Both conpounds showed the same effect in improving learning and memory， increasing Bmax of M-cholinergic receptors and accelerating cerebral protein and acetylcholine biosynthesis．However，Rgl but not Rbl had immunoregulatory action in aged rats and anti-osteoporosis effect in ovariectomized rats as well as enhanced basic synaptic transmission and magnitude of LTP induced by HFS． On the other hand，Rbl had anti-stress effects in antagonizing acute,chronic and repeated stress induced reduction of sexual behaviour and decrease of plasma andogen or estrogen．Rgl showed no such effect even aggravate stress induced damage．Rhl possessed anti-oxidant activity and prolong survival time of mice in cold(－10℃) condition．There was no any anti-cold effect with Rgl .These diference of biological activities between Rgl and Rbl may be arributed to their structures containing different number of glucoses．