| Ischemic cerebrovascular disease seriously influences to human survival quality,and The treatment principle is the timely restoration of blood perfusion in cerebral ischemia. However, ischemia and reperfusion can further increase the dysfunction and structural damage caused by ischemia, which the phenomenon is the cerebral ischemia reperfusion in jury. Reducing ischemia reperfusion damage and promoting their recovery has become an important research direction of the treatment of stroke disease.Recent studies show that many herbs have protective effect on cerebral ischemia reperfusion injury.The mechanism is mainly scavenging free radicals and reducing cellular calcium overload and toxicity of excitatory amino acids, inhibiting the inflammatory response and so on. This research discussed TCM protection function of ischemia reperfusion from cellular,molecular,gene level.It provided experimental basis for the mechanism of the Chinese medicine.ObjectiveTo observe the effect of Xuesaitong injection to OGD/R injured SH-SY5Y cells and try to clarify its intervention mechanism which can provide a scientific basis for its clinical application.Methods(1)The Non-toxic range of the Xuesaitong Injection on human neuroblastoma SH-SY5Y cells:We used MTT method to determinate the Non-toxic range of the Xuesaitong Injection on SH-SY5Y cells. In non-toxic range, we chose drug concentration according to certain proportion.(2)The protection of Xuesaitong Injection on OGD/R SH-SY5Y cells:we observed Xuesaitong injection anti-injury function on OGD/R SH-SY5Y cells.(3)The influence of Xuesaitong injection on OGD/R SH-SY5Y cells NogoA mRNA express ion:NogoA mRNA expression was detected by the real-time fluorescence quantitative.we clarified the mechanisms of Xuesaitong from the gene level.(4) The influence of Xuesaitong inject ion on OGD/R SH-SY5Y cells NogoA protein and its receptor expression:NogoA protein and its receptors expression were detected by immunocytochemistry. we clarified the mechanisms of Xuesaitong from the cell and the protein levels.In NogoA protein and its receptor expression experiments,There were eight groups:①ormal Control group,②oxygen and glucose deprivation reperfusion group (OGD/R),③OGD/R+NogoA antibody group,④OGD/R+NogoA antibody+Xuesa i tong group (divided into 320mg/L and 640mg/L group),⑤OGD/R+NogoR antibody group,⑥OGD/R+NogoR antibody+Xuesaitong group (divided into 320mg/L and 640mg/L group),⑦OGD/R+P75 antibody group,⑧OGD/R+P75 antibody+Xuesaitong group(divided into 320mg/L and 640mg/L group).We analysed the data by using computer image analysis system. Results(1)Xuesaitong injection intervened SH-SY5Y cells for 24h,48h,72h.We found that 320mg/L,640mg/L concentration has a good effect on the proliferation.We chose these two concentrations in this experiment.(2)SH-SY5Y cells'activity was significantly decreased (P<0.01) in the oxygen and glucose deprivation 3h and reperfusion 24h.Xuesaitong injection two concentrations had protective function on OGD/R injured SH-SY5Y(P< 0.01),and the high concentration had better effect than the low concentration(3)NogoA mRNA was significantly increased after OGD/R by using Real-time fluorescence quantitative method (P<0.01). NogoA mRNA expression decreased after we using NogoA antibody and Xuesaitong injection (P<0.01).(4)After SH-SY5Y OGD/R,NogoA protein and its receptor NogoR, P75 expression increased significantly(P<0.01).When we gave the corres-ponding antibodies and the Xuesaitong injectioin, their expression decreased (P<0.01).Conclusionthis experiment used SH-Y5Y lines and the three gas incubator to make oxygen and glucose deprivation model. The experimental results were stable and reliable. Studies had shown that injection of Xuesaitong had protective effect on OGD/R injured SH-SY5Y cells. It's mechanism may be as follows:It inhibited NogoA protein and its receptor NogoR,P75 expression; while reducing NogoA mRNA expression. |
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