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The interaction of Copolymer-1 induced T cells and microglia in RGCs survival protection and probable related immune mechanisms |
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| The interaction of Copolymer-1 induced T cells and microglia in RGCs survival protection and probable related immune mechanisms |
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作者：唐雅婷 文章来源：Department of Ophthalmology, Eye Ear Nose and Throat Hospital of Fudan University,Shanghai 200030, China 点击数：210 更新时间：2012/9/13 
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Aims Co-culture of Copolymer-1(Cop-1)-activated T-cells and microglia was made to observe its effect on retinal ganglion cells (RGCs) apoptosis in vitro and probable immune mechanisms was explored.
Methods We made co-culture of Cop-1-activated T-cells (Tcop-1) by repeated Cop-1 stimulation and microglia from retina of newborn rats in vitro and then the supernatant was collected. RGCs collected from retina of adult rats were purified and cultured for 10days. Then we added the supernatant of co-cultured Tcop-1 and microglia to RGCs (Tcop-1+MG group) to observe the RGCs apoptosis rate by Tunel analysis after 48h and compared it with other controlled groups (Tcop-1 group and MG group). RT-PCR was used to test mRNA change of related apoptosis protein (Caspase-3 and Caspase-8). The levels of probable cytokines including IGF-1, BDNF, TNF-a and IL-10 inthe supernatant of co-cultured Tcop-1 and microglia were also detected and compared with other matched groups in order to explore the possible molecular mechanisms that Tcop-1 and microglia interacted and worked on RGCs survival. One-way ANOVA and Boffironni method were used in the statistical analysis.
Results The RGCs apoptosis rate in Tcop-1+MG group was 25.36% after 48h, less than that of Tcop-1 group (31.03%) and MG group (61.54%) in Tunel test. RT-PCR results showed that the expression of mRNA of apoptosis protein Caspase-3 and Caspase-8 inTcop-1+MG group were 6.22×10-3 and 1.26×10-5 respectively, significantly lower than the mRNA expression of Tcop-1 group and MG alone group (both P<0.05). After co-culture of Tcop-1 and MG (Tcop-1+ MG group) for 12h, 24h and 48h, the secretion of IL-10 and BDNF in the supernatant were higher than that of T-cells and MG alone (P<0.05). The secretion of IGF-1 inTcop-1+ MG group also increased significantly and were more than the controlled groups after 24h and 48h(P<0.05). The secretion of TNF-a in Tcop-1+ MG group showed an increase after 12h compared with controlled groups but did not show statistical significance after 24 and 48h.
Conculsion The intereaction of Tcop-1 with microglia could delay the RGCs apoptosis process, showing the protection effect to optic neuron survival. The related immune mechanisms were complicated, but some clues including overexpression of BDNF, mediation and balance of some inflammatory cytokines (such as TNF-a) and anti-inflammantory cytokines (such as IL-10) are involved in the neuroprotection action. |
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