The Effect Of Danshen On HSP70,Bcl-2,Bax During Spinal Cord Ischemic Reperfusion Injury

Objective:Observing the Danshen and Tanshinone on ischemia reperfusion injury in spinal cord by neuronal apoptosis relating protein HSP70, Bcl-2, Bax expression, and further explore the protection mechanism of the Danshen on spinal cord ischemia reperfusion injury.Methods:120SD rats weighting250±10g of clean grade,were random divided into Sham operation group, Ischemia reperfusion group, Danshen group and Tanshinone group,30rats in each.The ministry of right renal artery without occlusion of the abdominal aorta after the operation is terminated in the Sham group. Using the model improved and estanblished by Li Zhang in the Ischemia reperfusion group,giving no drug intervention before surgery.Danshen injection intraperitoneally half an hour before the spinal cord ischemia reperfusion injury model estanblished in the Danshen group.Tanshinone group inject tanshinone Ⅱa sodium injection intraperitoneally half an hour before operation. Each group derived after modeling0.5h,1h,4h,8h,12h,a total of five time points,6rats at each time point.The HSP70, Bcl-2, Bax levels of Spinal cord tissue was detected by ELISA, and the Bcl-2, Bax positive cells of Spinal cord tissue was detected by immunohistochemical staining. The pathological changes of spinal cord tissue are observed under light microscope through Nissl staining.Results:1The expression of HSP70in the Sham group was slightly at each time point, the expression of HSP70increased at0.5h in the Ischemia reperfusion group,and then increasing the expression gradually over time in12h.The Ischemia reperfusion group compare with the Sham group was significant difference at the same time point(P<0.01);The expression of HSP70in the Danshen group and the Tanshinone group were higher than the Ischemia reperfusion group at each time point, the three were compared with the very significant difference (P<0.01).2The expression of Bcl-2is low in Sham group at each time point,the expression of Bcl-2in the Ischemia reperfusion group increased from1h and then increase expression gradually.The expression of Bcl-2in the Ischemia reperfusion group compare with the Sham group was significant difference at the same time point(P<0.01);The expression of Bcl-2in the Danshen group and the Tanshinone group compare with the Ischemia reperfusion group were higher with very significant difference at the same time point(P<0.01).3The expression of Bax in Sham group is very small at each time point,but the expression of Bax in the Ischemia reperfusion group increased from0.5h,and then increased gradually in12h. The expression of Bax in Ischemia reperfusion group compared with the Sham group were significant difference at each time point (P<0.01);The expression of Bax in the Danshen group and the Tanshinone group are lower than the Ischemia reperfusion group with very significant difference at each time point(P<0.01). 4The Sham group do not have significant pathological changes in spinal cord tissue at very time points. You can see part of the reducing size or mild swelling of neurons after injurying0.5h,1h. Nissl bodies reduce slightly with no significant changes in the nucleus;The pathological changes increased with the time goes.The cell morphology and the Nissl bodies in the Danshen group and the Tanshinone group were always better than the Ischemia reperfusion group at each time point.Conclusions:1In this experiment,the rat model of spinal cord ischemia reperfusion injury is highly feasible and reproducible,it can meet the needs of experimental model of disease, should be widely applied.2In the spinal cord ischemia reperfusion injury, the Danshen and Tanshinone can reduce neuronal apoptosis to protect the spinal cord by promoting the expression of HSP70,and then promoting the expression of Bc1-2to inhibit the expression of Bax.3It can be confirmed that the Danshen and Tanshinone can protect spinal cord neurons after spinal cord ischemia reperfusion injury by pathological changes.