Effects Of Pilose Antler Polypeptide Combined With GDNF Gene Modified Schwann Cells On Radiation Induced Spinal Neurons Apoptosis

Purpose: To investigate the effect of Pilose antler polypeptide combined with GDNF gene modified Schwann cells on the expressions of Bax protein and Bcl-2 protein as well as apoptosis rate after radiation induced spinal neurons apoptosis.Method: The spinal neuron suspension of fetal rats was prepared.The spinal neurons in logarithmic growth phase were taken and the apoptosis was induced by radiation.The cultured cells were divided into the six groups.A group: normal spinal neurons group;B group: radiation induced apoptotic spinal neurons culture group;C group: radiation induced spinal neurons apoptosis + SCs group;D group: radiation induced spinal neurons apoptosis +pEGFP-N1 transfected SCs group;E group: radiation induced apoptotic spinal neurons +pEGFP-N1/GDNF transfected SCs group;F group: Pilose antler polypeptide +pEGFP-N1/GDNF transfected SCs + radiation induced apoptotic spinal neurons group.The apoptosis rates of all groups were detected using the flow cytometry.The expressions of Bcl-2protein and Bax protein in spinal neurons were observed by immunohistochemistry.Result: The spinal neuron suspension of fetal rats was prepared.The spinal neurons in logarithmic growth phase were taken and the apoptosis was induced by radiation.The cultured cells were divided into the six groups.A group: normal spinal neurons group;B group: radiation induced apoptotic spinal neurons culture group;C group: radiation induced spinal neurons apoptosis + SCs group;D group: radiation induced spinal neurons apoptosis +pEGFP-N1 transfected SCs group;E group: radiation induced apoptotic spinal neurons +pEGFP-N1/GDNF transfected SCs group;F group: Pilose antler polypeptide +pEGFP-N1/GDNF transfected SCs + radiation induced apoptotic spinal neurons group.The apoptosis rates of all groups were detected using the flow cytometry.The expressions of Bcl-2protein and Bax protein in spinal neurons were observed by immunohistochemistry.The apoptosis rate testing results were as follows: Compared with the normal spinal neurons group,the apoptosis rate in the radiation induced spinal neurons culture group was significantly increased(p<0.05);Compared with the radiation induced spinal neurons culture group,the apoptosis rates of radiation induced spinal neurons culture + SCs group,radiation induced spinal neurons apoptosis + pEGFP-N1 transfected SCs group and Pilose antler polypeptide +pEGFP-N1/GDNF transfected SCs + radiation induced apoptotic spinal neurons group were decreased(p<0.05);Compared with the apoptosis rate of radiation induced spinal neurons culture + SCs group,the apoptosis rates in the radiation induced apoptotic spinal neurons +pEGFP-N1 transfected SCs group and Pilose antler polypeptide + pEGFP-N1/GDNF transfected SCs + radiation induced apoptotic spinal neurons group were decreased significantly(p<0.05).Immunohistochemistry result showed that Bcl-2 positive cells were brown.The results showed that Bcl-2 was expressed in A,B,C,D,E and F groups.The Bcl-2positive cells were minimum in A group and the staining was slight.The number of Bcl-2positive cells was maximum in F group and the staining was deepest,and the staining showed significant difference between F group and C,D and F group.The number of positive cells and staining showed no significant difference among C,D and E groups.Thus,the expression of Bcl-2 was increased in Pilose antler polypeptide + pEGFP-N1/GDNF transfected SCs +radiation induced apoptotic spinal neurons group,thereby inhibiting the apoptosis of spinal neurons.The Bax was expressed in A,B,C,D,E,F groups.The number of Bax positive cells was lowest in A group and the staining was slight.The number of Bax positive cells was maximum in B group and the staining was darker.Compared with C,D and E groups,the number of Bax positive cells in F group was much less and the staining showed significant difference.The number of Bax positive cells showed no significant difference among in C,D and E groups and the staining had little difference,but compared with the B group,which was decreased significantly.It can be seen that the expression of Bax was decreased in the Pilose antler polypeptide + pEGFP-N1/GDNF transfected SCs + radiation induced spinal neurons apoptosis group,thereby inhibiting the apoptosis of spinal neurons.Conclusion: Flow cytometry results showed that the apoptosis rates of radiation induced apoptotic spinal neurons + Pilose antler polypeptide group and radiation induced apoptotic spinal neurons + pEGFP-N1 transfected SCs group were lower than those of radiation induced spinal neurons culture group and Pilose antler polypeptide group + radiation induced apoptotic spinal neurons + pEGFP-N1/GDNF transfected SCs group were significantly lower than that of the radiation induced apoptotic spinal neurons + pEGFP-N1 transfected SCs group,but higher than that of normal spinal neurons group.So it was inferred that compared with the Schwann cells in the Pilose antler polypeptide group or GDNF modified Schwanncells group,Pilose antler polypeptide + GDNF modified Schwann cells may have a better advantage.