The Effect Of Tanshinone To GLU,GSH,ATPases During Spinal Cord Ischemic Reperfusion Injury

Objective:Exploring the protective effect and mechanism of tanshinone during rat spinal cord ischemia-reperfusion injury by observing the rat spinal cord neurons glutamate (GLU) levels, serum glutathione (GSH) content, spinal cord or nerve cell membrane Na+-K+-ATP enzyme activity.Methods:250±10g body weight of SD rats were randomly divided into Tanshinone group, ischemia-reperfusion group and sham-operated group of 40. Sham group, anesthesia and opened the abdominal cavity, but not occlusion of the abdominal aorta to the termination of surgery, the temporary closure of abdominal cavity; Ischemia-reperfusion group:not injecting any medication, close the abdominal cavity after modeling; Tan Group: intraperitoneal injection of sodium tanshinoneⅡa injection a half-hour before spinal cord ischemia-reperfusion model produced and then close the abdominal cavity. In each group were taken at the anesthesia abdominal aortic blood, after modeling 0.5h, 1h,4h,8h,12h time points of serum glutathione, spinal cord glutamate, Na+-K+-ATP activity and observe the morphological changes of nerve cells by light microscopy.Results:1 After the spinal cord reperfusion,the glutamate of Sham group and ischemia-reperfusi-on group, Tan group began to increase,from 429.62,643.63,518.00umol/gprot in 0.5h, reached the highest point:440.00,1308.38,581.92 umol/gprot in 4h,and then gradually decreased,12h achieve 445.25,498.25,451.75 umol/gprot, did not reach the normal level,compared three groups at the same time a very significant difference (P<0.01); ischemia-reperfusion group compared with the Tan group at the same time has very significant differences (P<0.01).2 After the spinal cord reperfusion,the serum levels of glutathione content of Sham group and ischemia-reperfusion group, Tan group started to decline,from 21.08,10.05,19.57 mgGSH /L in 0.5h, reached the lowest point:18.09,1.03,16.34 mgGSH/L in 4h,and then gradually increased,12h achieve 19.41,6.10,13.72 mgGSH/L, did not reach the normal level,compared three groups at the same time a very significant difference (P<0.01); ischemia-reperfusion group compared with the Tan group at the same time has very significant differences (P<0.01).3 After the spinal cord reperfusion,the Na+-K+-ATP activity of Sham group and ischemia- reperfusion group, Tan group started to decline,from 5.87,5.30,5.67 umolPi/mgprot/hour in 0.5h, reached the lowest point:6.00,2.00,4.04 umolPi/mgprot/hour in 4h,and then gradually increased,12h achieve 7.82,4.05,6.06 umolPi/mgprot/hour, did not reach the normal level,compared three groups at the same time a very significant difference (P<0.01); ischemia-reperfusion group compared with the Tan group at the same time has very significant differences (P<0.01).4 Tanshinone group and ischemia-reperfusion group of nerve cells began to appear addicted in 0.5h increased silver-colored, and other changes to enhance the deep changes, only a small amount of degeneration of nerve fibers,4h degeneration of nerve cells around the peak, we can see clusters of axon degeneration,protruding into a black bead-like arrangement of samples, or disconnected, neuron cell body swelling, cytoplasm containing granular deposition of silver particles, a number of cell death, the formation of vacuoles, particularly in ischemia-reperfusion group was significantly more reperfusion 8h,12h no obvious nerve cells to further degeneration and necrosis, degeneration and necrosis of cells, axons and so no significant expansion of the scope.Conclusions:1 The spinal cord ischemia model used in this study is stable, feasible, reproducible, and can meet the requirements of the experiment.2 In the spinal cord ischemia-reperfusion injury during the spinal cord, can delay content of GLU increasing,serum GSH content decreasing and Na+-K+-ATP activity decreasing so as to influence EAAs-Xc-system. pathway and thus to reduce glutamate-induced neuronal toxicity.3 Tanshinone can delay the spinal cord tanshinone content of GLU increasing, serum GSH content decreasing and the Na+-K+-ATP activity decreasing during the spinal cord ischemia-reperfusion injury,reduce glutamate-induced neural cell toxicity by affecting the EAAs-Xc-system pathway.