The Effect Of Egb761 And NO On Apoptosis,Free Radicals And Bcl-2 Of Hypoxia Neurons

Neurons apoptosis after cerebral ischemia is a kind of PCD(programmed cell death), It mainly occurs in the IP( ischemic penumbra), which is the decisive factor in prognosis of cerebral ischemia disease. Some experiments indacated that NO(nitric oxide) had neurotoxicity and was connected with hypoxia-induced neurons apoptosis, however, its mechanism is still not clear. In our research, we studied the role of NO in hypoxia-induced neurons apoptosis of rats and its mechanism concerned with bcl-2 and oxidative free radicals. Then studied the effect in this mechanism of Egb761 on hypoxia-induced cortical neurons apoptosis, using serum and cerebrospinal fluid pharmacological test. We hope our research can provide theoretic and experimental evidence for its exertion.METHODS: We cultured the cortical neurons of fetal rats, treated with hypoxia to establish experimental model. Effects of Egb761 and NO on hypoxia-induced cortical neurons apoptosis was assessed using MTT test and flow cytometric DNA analysis; We assayed the level of MDA and NO in medium to study the effects of Egb761 on the oxidative free radicals and NO produced by these apoptosis neurons. The level of bcl-2 expression was analysized by flow cytometry.RESULTS: Cultured cortical neurons treated with hypoxia undergo apoptosis, demonstrated by typical DNA ladder and apoptic bodys under electron microscope; Apoptosis rate and MDA in high NO group were significantly higher than the control (P<0.01), however, bcl-2 in this groupwas down-regulated, compared with the control P<0.01; NO level and MDA and apoptosis rate in Egb761 group were significantly lower than the control (P<0.01), bcl-2 expression in this group was up-regulated, compared with the control P<0.01.CONCLUSION: NO can enhance hypoxia-induced cortical neurons apoptosis via down-regulating bcl-2 expression and transforming into oxidative free radicals. Egb761 has protective effects on hypoxia-induced neurons apoptosis, its mechanism may be concerned with restraining the NO and oxidative free radicals , and up-regulating bcl-2 expression.