| Objectives1. To explore the establishment of myelin proteolipid protein induced-EAE animal model and its relationship with the animal gender.2. (l)To explore the proliferation of microglia and astrocyte and its relationship with the clinical course of EAE.(2) To explore the expression of chemokines of IP-10, MCP-1, MIP-1 a and MIG, etc in CNS of mouse , and their relationship with the clinical course of EAE. (3)To observe the expression of FasL in the CNS of mouse and to explore the relationship with the clinical course of EAE.3. To explore the therapeutic potential of Qingkailing for EAE and its mechanism.Methods1. Each SJL mouse was injected s. c. over the abdominal wall with PLP139-151,Mycobacteria H37RA and IFA, and Bordetella pertussis bacilli.2. The expression of GFAP, RCA-1, FasL and the chemokines of IP-10, MCP-1, MIP-1 a , MIG in the CNS of SJL mouse were studied with using immunohistochemistry technique.3. EAE was induced by PLP139-151 in SJL mouse, all mice were weighted and examined daily for neurologic signs, the secretion of interferon- Y and the proliferation response of lymphoid cells induced by PLP139-151 were detected.4. The effect of Qingkailing on the expression of CD3,CD4,CD8 and CCR5 of the lymph node(LN) cells and splenocyte in EAE mice was studied by using flow cytometry technique.Results:1. Incidence of EAE in immunized female animal reached 80%, most mice developed clinical EAE within 15?.1 days of immunization. Myelin PLP irnmunostaining shows a large unstained demyelinated area in spinal cord of immunized mice. Moreover, The female animal model of EAE induced by PLP had features of stability, relapse-remitting and high incidence. Incidence of EAE in immunized male animal also reached more than 80%, However, most male mice developed clinical EAE within 22 +4. 3d days of immunization, which displayed a monophasic episode without relapse-remitting feature.2. (1) There existed the activation and proliferation of astrocyte and microglia in the whole EAE course, whichthe activation of astrocytes was the most obviously displayed in the remitting course of EAE. However, the activation of microglia in acute episode of EAE was most obviously.( 2 ) The expression of the chemokines of IP-10, MCP-1, MIP-1 a and MIG in the CNS of mouse were increased siginificantly in the acute and relapse episode of EAE,but were decreased in the remitting course. Furthermore, the morphologic features of IP-10, MCP-1 positive cells seemed to be similar with that of astrocyte, the morphologic features of MIP-1 a and MIG positive cells seemed to be similar with that of microglia.(3)The expression of FasL in CNS of mouse was siginificantly increased in the whole course of EAE mouse, and the increasement of the number of FasL-positive neuron reached at the top level.3.Qingkailing can significantly inhibit the secretion of interferon- Y and proliferation response of lymphoid cells induced by PLP139-151, Mice received Qingkailing treatment had a significantly improved clinical outcome compared with control group(p<0.01).4.The percentage of CDS and CDS positive cells of LN cells and splenocyte in mice received Qingkailing treatment was significantly increased, however the percentage ofCD4/CD3 and CCR5 and the ratio of CD4/CD8 was significantly decreased compared to the control group. Mice received Qingkailing treatment had a significantly improved clinical outcome compared with control group (p<0. 01).Conclusion:1. The female animal model of EAE induced by PLP139-151 had features of stability, relapse-remitting and high incidence, which was considered a better model for the study of multiple sclerosis.2. (1)The activation of astrocytes and microglia may affect the pathologic process of EAE , in part by regulating the expression of chemokines of IP-10, MCP-1, MIP-1 a and MIG, which may be involved in the pathogenesis of EAE and MS. (3)The neuron apoptosis may be an important feature in the pathophysiologic process of EAE.3. Qingkailing...
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