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Ginsenoside Rd Attentuates Ischemic Stroke-induced Inflammation in Rats

Author: ZhangYunXia
Tutor: ZhaoGang;ShiMing
School: Fourth Military Medical University
Course: Neurology
Keywords: Ginsenoside Rd Inflammatory response Microglia Cytokines Liquid chip
CLC: R285.5
Type: Master's thesis
Year: 2011
Downloads: 43
Quote: 0
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Ischemic cerebrovascular disease, also known as stroke, has become the second largest cause of death in the world, because of its pathogenic mechanisms are complex, affecting a wide range of clinical few clear neuroprotective drugs. Ginseng and Panax is our application of a wide range of traditional medicines, medication history for thousands of years has proved its efficacy and safety. As one of the two drugs effective active ingredients, ginseng saponins (Ginsenosides, GS) has been shown to reduce brain infarct volume after ischemia-reperfusion, protection of hippocampal pyramidal neurons. Determine ginsenosides effective monomer component of its mechanism of action, which is the primary task to develop a new type of brain protection drugs. Ginsenoside Rd (Ginsenoside Rd, GSRd) as the main active ingredient of ginsenosides one has a variety of pharmacological effects of resistance convulsions, anti-aging, regulate immune. Our previous studies showed that GSRd can protect PC12 cells by inhibiting hydrogen peroxide toxicity injury; inhibition of oxidative stress injury caused by oxygen and glucose stripping model; inhibition of intracellular calcium overload in neurons against glutamate excitotoxicity . The clinical trial results also confirmed GSRd effective treatment for patients with acute ischemic stroke. These findings indicate that GSRd has significant neuroprotective effect, but the mechanism is not yet fully understood. Studies show that the inflammatory response to cerebral ischemia and reperfusion injury, involving a variety of immune cells and immune factors, new direction to reduce ischemic injury by inhibiting the inflammatory response and cytokine production of brain protection research . After cerebral ischemia microglia (microglia, MG) on the one hand can be used as antigen presenting cells, mediated by the inflammatory response of the damaged area; other hand, they can secrete a variety of substances to regulate the inflammatory response. GSRd will impact the status and function of the central nervous system of the MG to play a neuroprotective effect has not been determined. Experiment first observed the the inflammatory response GSRd of MG, then gradually in-depth study GSRd explore cerebral ischemia GSRd mechanisms play an anti-inflammatory effect on the role of a variety of cytokines secreted by MG. Experiment on cerebral ischemia-reperfusion injury after MG Objective: Research GSRd on cerebral ischemia-reperfusion injury a GSRd MG secrete a variety of cytokines. Methods: middle cerebral artery occlusion recanalization simulated cerebral ischemia-reperfusion injury, respectively reperfusion after 12 hours, 1 day, 3 days and 7 days of the four time points MG immunohistochemical staining was observed in morphology and changes in the number GSRd (10mg/Kg). Using semi-quantitative PCR after cerebral ischemia GSRd (10mg/Kg) the NF-κB mRNA level changes. Using liquid chip to detect cerebral ischemia, and each time point using GSRd (10mg/Kg) IL-1β, IL-6, IL-18, TNF-ɑ, IFN-γ, of CXCL1, TGF-β protein levels . Results: immunohistochemical staining results showed that cerebral ischemia and reperfusion injury, MG significant activation increase in the number, morphology rapid activation state by ramified resting state, and its degree of activation in a time-dependent manner, in the first 7 days to reach the peak; using the GSRd pretreatment MG each time point, significantly reduced the number, the activated cell size smaller. The Luminex test results show that the cerebral ischemia, IL-1β, IL-6, IL-18, TNF-ɑ, IFN-γ, CXCL1 protein content was gradually increased, and the third day peaked TGF-β levels increased; GSRd pretreatment, IL-1β, IL-6, IL-18, TNF-ɑ, IFN-γ, CXCL1 RNA and protein content significantly decreased, TGF-beta was significantly increased. The semi-quantitative PCR showed the same trend of NF-κB mRNA. Conclusion: GSRd can reduce the number of MG activation, inhibition of the the MG secretion of IL-18, TNF-ɑ, to increase the content of TGF-β, resist inflammation in cerebral ischemia reperfusion injury. The prompted GSRd inhibit cytokine production, reduce the inflammatory response may play a neuroprotective effect mechanism. The second experiment ginsenoside Rd on primary cultured MG Objective: observed cultured MG and secrete a variety of cytokines in primary GSRd activated by lipopolysaccharide (lipopolysaccharide, LPS). METHODS: Primary cultured neonatal rat MG. The immunofluorescence staining MG purity. To 12 days in vitro culture experiments. To use LPS activation MG, observe the use of 10μM GSRd pretreated MG morphology and number changes; Luminex detection LPS activation and 10μM GSRd pretreated cells supernatant IL-1β, IL-6, IL-18 , TNF-ɑ, IFN-γ, CXCL1 protein levels. Results: no significant difference between normal cells and 10μM GSRd of the control group; administration of LPS activation MG MG morphology from rod into amoeba-like, while the number of activated cells significantly increased; while using 10μM GSRd pretreatment after amoeba like MG reduce the number of the cell size smaller. The the Luminex test results also show that no significant difference between the control group of normal cells and 10μM GSRd of LPS activation of IL-1β, TNF-ɑ, IL-6, IL-18, IFN-γ, of CXCL1 content increased significantly, the use of The content of these cytokines was significantly reduced the 10μM GSRd after pretreatment, but higher than that in the control group, the difference was statistically significant. Conclusion: GSRd can be reduced MG number and volume of the LPS-activated, suppressed MG secrete IL-1β, IL-6, IL-18, TNF-ɑ, IFN-gamma, CXCL1, Step GSRd by inhibiting the secretion of cytokines affect MG morphology and function, to reduce inflammatory response, and have neuroprotective effects.

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