| It is a worldwide subject for therapy of spinal cord injury (SCI). Although there have been taken amazing outcome on gene therapy of SCI in experiment study,for the complex effect factor and injury mechanism,there is still a lot of key point unknown for people to study incontinuous.TNF-αis a cytokine of inflammation, which can be released from neuron and others.In this research, we will detect the protection role of monoclone TNF-αAntibody in spinal cord injury.In this research, the BBB achievement of the SCI group and TNF-αtreated group were lower than that of the sham group at after 12 h, 1 d after SCI.the BBB achievement of the SCI group and TNF-αtreated group were lower than that of the sham group at 3 d, 7 d, and 14 d after SCI,While at 7 d after SCI, compared with sham group, the BBB of TNF-αtreated group was higher significantly than that of SCI group( P < 0.05).While at 14 d after SCI, compared with SCI group, the BBB of TNF-αtreated group was higher significantly than that of SCI group( P < 0.05).Histology detection found that morphorlogy of spinal cord nerve cell of the sham group was normal. spinal cord nerve cell edema can be seen, but no apoptosis can be seen at 12h after SCI,inflamatory cell infiltration can be seen in spinal cord tissue at 3d after SCI,repair change can be seen in spinal cord tissue at 7d after SCI,scar formation can be seen in injured spinal cord tissue at 14d after SCI,while at 7d after anti- TNF-αtreatment,nerve cell morphology begin to repair, and function of spinal cord. at 14d after anti- TNF-αtreatment, spinal cord tissue structure arrange regular, inflammatory cell disappear.The immunochemistry results of Image analysis of bcl-2 and bax showed that the index of bcl-2/bax of SCI group decresed significantly at 12 h after SCI, compared with the shamI group, P<0.05. which increase gradually.While at 12h,1d,3d,7d after anti-TNF-αtreatment, the index of bcl-2/bax of TNF-αtreated group increased significantly, compared with SCI group, P<0.05.The RT-PCR results show that bcl-2 mRNA expression level of SCI group increase at 12h after SCI, reach the peak walue at 24h after SCI, compared with sham group, P<0.05. then which decrease gradually, but which is higher compared with sham group, P<0.05. After treated with anti-TNF-αantibody, the bcl-2 mRNA expression of TNF-αtreated group was higher significantly than that of SCI group(P<0.05)at12 h,1d,3d,7d after SCI.the bax mRNA expression was increasing and reach the peak value at 12h after SCI, compared with sham group, P<0.05.and then decreased.the bax mRNA expression of SCI group was higher significantly than that of sham group(P<0.05)at 1d,3d,7d after SCI.while anti-TNF-αtreated was applied, bax mRNA expression of TNF-αtreated group was no significantly difference from that of SCI group at each time point. (P﹥0.05)Western blot results show that,the bcl-2 protein expression was increasing at 12 h after SCI and reach the peak value at 24h after SCI, and then decreased. compared with sham group, P<0.05.While anti-TNF-αtreated was applied, bcl-2 protein expression of TNF-αtreated group was significantly higher than that of SCI group, compared with SCI group at each time point., P<0.05.bax protein expression of SCI group was at the peak value at 12h after SCI, and decreased furtherly at 1d,3d and 7d after SCI, compared with sham group, P<0.05.After treated with anti-TNF-αantibody, the bax protein expression of TNF-αtreated group was no significantly difference from that of SCI group at each time point. (P﹥0.05)ELISA analysis found thatat 12 h after SCI, amount of IL-6 and IL-8 expression was at the peak value, compared with the sham group, P<0.05.with times go on, the amount of IL-6 and IL-8 expression were lower grandually than that of SCI group, and at 14 d after SCI, the amount of IL-6 and IL-8 expression were access to the normal level.While after anti-TNF-αtreatment was applied, IL-6 and IL-8 expression of TNF-αtreated group decreased significantly, compared with SCI group, P<0.05. TUNEL detection was applied to evaluate the apoptosis in the SCI rats. It was found that the apotosis cell can be seen clearly 3 days after SCI, which can be prevented after treatment of anti-TNF-α.From our research, we can get these conclusions:1. apoptosis is an important event of SCI; 2. Bax/bcl-2 family take part in the development of SCI through control the processof apoptosis; 3. TNF-αplay an important role in SCI, especially in regulating signal transduction pathway of apoptosis; 4. Anti-TNF-αtreatment by monoclone antibody can regulate the expression of IL-6,IL-8 and bcl-2,inhibit the process of apotosis after SCI in rats. |
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