Investigate The Effect Of Inflammatory Factors On Cultured Trigeminal Ganglion

Objective The integral trigeminal ganglia(TG) of rat can be cultured to set up a biological activity system similar to internal environment which could be easy controlled during experiment. Investigate the effect of inflammatory factors on CGRP levels of cultured trigeminal ganglion.Methods 18 adult male SD rats were acutely isolated to get the 36 TG. Among them, 12 TG were fixed by 4% paraformaldehyde, the other 24 TG were cultured. Those 24 TG were averaged divided into control group and experimental group after culturing for 6 days. In control group, the culture solution was changed by normal fresh culture solution on the 6th day. In experimental group, the culture solution was changed by normal fresh culture solution with TNF-α on the 6thday. Then, the Cellular morphology was detected by HE staining, and the calcitonin gene-related peptide(CGRP) positive stained cells were detected by immunohistochemistry staining and Western Blotting to examine the effects of TNF-α, a kind of proinflammatory cytokines, on CGRP expression.Results 1. The HE staining results show that there has no significant difference of both fresh TG and explanted culture TG in tissue edema, degeneration, necrosis and apoptosis. 2.Compared with the control group, the expression of CGRP receptors in TG neurons of treatment groups is significantly increased(P < 0.01) with Western Blotting detection. This indicates that inflammatory cytokines and CGRP might be very important in the research of trigeminal neuralgia(TN). 3. TG neurons of rat detected by immunohistochemistry show CGRP positive staining in control group and experimental group. Compared with the control group, CGRP positive stained cells in TG neurons of treatment group are significantly increased, and cells appear brown, cell bodies dye two-tone, some cells near the center appear shading light and the surrounding cytoplasm appear deeper. 4. Compared with the control group, expect the average area, mean density, integral optical density,average gray of TG slices are significantly increased with immunohistochemical detection in treatment groups(P < 0.01).Conclusion Inflammatory cytokines, TNF-α, can significantly increase CGRP levels of integral TG of rat in culture condition. Culturing integral TG of rat is technically available from morphological analysis, the internal environment of cultured TG is easy to control,and morphological and molecular biological detection is easy to carry out. Therefore,cultured integral TG is an ideal model in the functional mechanism research of TG and its relative drug studies.