| Obejective(1) To study the neuroprotective effect of salidroside in MCAO rats; (2) To study the effects and partial mechanisms of inhibite complement signaling pathway on salidroside neuroprotection.Methods(1) 63 Male sprague dawley rats are administrated with salidroside for 6 days after lh of the ischemia reperfusion. Zea-Longa neurological deficit score, Nissl’s staining, Gene chip technology, RT-PCR, Western blot and fluorescent technique are used for detection.(2) 36 Male sprague dawley rats are administrated with salidroside for 6 days after 48h of the ischemia reperfusion. Zea-Longa neurological deficit score, Nissl’s staining, Gene chip technology, RT-PCR, Western blot and fluorescent technique are used for detection.(3) To establish a hypoxic damage model of PC 12 cells induced by CoCl2 and detect the Caspase-3 activity change and the protein expression of Egr4, Egrl under the different concentration of salidroside.(4) To study C3a/C3aR signaling pathway on neuroprotective effect of salidroside,72 Male sprague dawley rats which undergo the MCAO are administrated with salidroside for about 1d,2d and 6d respectively for the detection of Neun, IgM, Annexin IV, C3 and MBL-2.(5) 72 Male sprague dawley rats are seperately injected into the left lateral ventricle C3ar inhibitor and artificial cerebrospinal fluid with the help of ventricle stereotaxic apparatus. 30 minutes later, the Models of MCAO are finished with 1 hour reperfusion, drug administration and intracerebroventricular injection for 2 days. At last, the expressions of Neun, IL-6, Egrs were detected.Result(1) The rats undergo 1 hour ischemia reperfusion with 6 days salidroside treatments show that salidroside can improve the neurological deficit score, increase the number of positive cell of Nissl,decrease the nuber of apoptotic cells, lower the proteinic level of cleaved Caspase-3 but no influence on total level, inhibit the expressions of Bax, NogoA and NgR, enhance the BDNF,NGF and Bcl-xL protein. The gene chip show salidroside can reverse the genomic change of ischemia reperfusion.(2) Simultaneously, the rats after 48-hour-reperfusion with the treatment of salidroside have a smaller infarct volume, better deficit score and promote the expression of gene and protein such as Egrl,Egr2,Egr4 and Arc.(3) Salidroside can inhibit the activity of Caspase-3 in model of PC 12, promote the proteinic expression of Egr4, Egrl. with the interference of Egr4 by siRNA,not only a more active Caspase-3 and an increasing expression of Bax has been found.but also the Bcl-xL was degraded. However, All of that can be reversed by the treatment of Salidroside.(4) The MCAO rats are administrated with salidroside for ld,2d and 6d respectively.it not only reverse the expressions of IgM, C3, Annexin IV and MBL-2, promoting the NeuN, but also reduce the C3 in neurocyte. It has no obvious effects on Clqa and Egr4, Arc in one-day treatment.but it can reduce the C1qa and enhance the expression of Egr4, Arc after 2 days.(5) The rats are administrated with C3aR inhibitor into cerebral ventricle continueously, it show the same result in accord with the treatment of salidroside. What is more, the treatment of salidroside and C3aR inhibitor have never shown the remarkable additive effects.Conclusion(1) Salidroside which has a neuroprotective effect with an advantage of wild therapeutic window can alleviate the infarct volume, improve the neurological deficit, inhibit the neural apoptosis and reverse the hypoxia-reperfusion genetic change.(2) Salidroside, a neuroprotective drug, can promote Egr4 expression,inhabite apoptosis,and have neuroprotection.(3) Salidroside can alleviate the death of neurocyte by inhibiting the complement system with a time and cell type dependence, promoting the NeuN by inhibiting Brain endothelial lectin pathway in acute phase. Though Egrs doesn’t work in acute stage, it is promoted by salidroside in lectin inhibition pathway and classic pathway of complement system. |
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