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Objective:On the basis of traditional chinese medicine, with the modern biotechnology to investigate the protective effect and its mechanisms of genistein sodium sulfonate (GSS), a derivatives of the effective component in pueraria, on brain ischemia/reperfusion injury in rats from different angles of oxidative stress, energy metabolism, excitatory toxic injury, inflammatory response and so on. It can provide the theroretical and practical foundation for expanding pharmacological effects of pueranae.Methods:A rat ischemia/reperfusion injury animal model was establish through embolization of the middle cerebral artery, and a cell injured models were induced by glutamate. Through different research methods like nerve function scores, TTC staining, enzymatic assay and test technology, radioooimmunoassay, enzyme-linked immuno sorbent assay, MTT method, flow cytometry, Western blot and so on, to study the protective effect and mechanisms of GSS on brain ischemia/reperfusion injury in vitro and in vivo.Result:(1) MCAO model rats showed significant injury in neurological function and infarction volume, increases in LDH and CK activities in serum while decreases in brain tissue. After treated with GSS, the neurological score, infarction volume and LDH and CK activities in serum were reducted, and the LDH activitiy in brain tissue were increased.(2) Compared with control rats, the rats suffered ischemia showed decreases in the activities of SOD, GSH-Px, CAT, tNOS and cNOS in the injuried brain tissue and similarly in T-AOC. But increases in activity of iNOS. Moreover, its content of NO is lowered while increased of MDA. After treated with GSS, these changes were transformed.(3) GSS can enhance the activities of Na+-K+-ATPase and Ca2+-Mg2+-ATPase on the affected brain.(4) GSS treatment significantly increased the viability of PC12cells injuried by glutamate.(5) The control primary culture cortical neurons have good refraction, body full and more dendrite meanwhile. The morphology and ratio of LDH release do not change in neurons by simple treated with GSS. The neurons injuried by glutamate showed worse refraction, body full, less dendrite and more LDH release, compared with the normal neurons. However, the resulted is opposite by GSS treatment. These dataes show that GSS can protect the cortical neurons against glutamate induced neurotoxicity in vitro.(6) GSS can derease the expression of GluN2B, a NMDAR subunit, in MCAO rats’ hippocampus.(7) Treated with GSS, the rats suffered cerebral ischemia/reperfusion injury showed an increase in IL-10content in serum and expression of IL-10and IL-10R in half injury area of cortex and hippocampus, while decreases in IL-6and TNF-α levels, and the activity of MPO in serum.(8) The protein of Bax, Caspase-3and Caspase-9are increased in the ischemic penumbra area of cortex, while Bcl-2is decreased. After the treatment by GSS, all that is reversed.(9) It’s showed that the protein expression of Bax and Caspase-3is increased in the injuried neurons but Bcl-2is down. These changes were transformed through treatment by GSS.(10) The protein expression of downstream regulatory element antagonist modulator(DREAM) is decreased in the ischemic penumbra area of cortex and hippocampus. After treated with GSS, the expression of DREAM in the same tissues was increased.Conclusion:(1) GSS had protective effects on brain ischemia/reperfusion injury in rats.(2) GSS can increase antioxidant capacity and then alleviat oxidative stress injury, improve the activity of ATP and then modulate the disorder of energy metabolism, antagonize the excitatory by regulating the expression and activation of NMDA receptor, inhibit inflammatory response and cell apoptosis to provide protective effects on brain ischemia/reperfusion injury in rats.(3) DREMAM may play an important role in protective effects on brain ischemia/reperfusion injury in rats.
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