Expression Of Brevican And Neurocan In Activated Astrocytes By The Effect Of Olomoucin

Objective To study the expression of brevican and neurocan in activated astrocytes bythe effect of olomoucin, and elucidate the role of them in activated astrocytes.Methods Astrocytes were separated and purified by shaking culture method combinedwith differential adhesion method, then the third generation of astrocytes were used tosubsequent experiment and divided into three groups, respectively for the control group,activation group and inhibition group. Astrocytes in the control group were culturedwith10%FBS DMEM culture medium, in the activation group were co-cultured with20ng/ml ciliary neurotrophic factor (CNTF), and in the inhibition group were incubated100μmol/L olomoucine. Cell morphology was examined using immunocytochemistry. Threegroups of cells by different conditions deal with12,24and48hours later,the content ofbrevican and neurocan were examined by sites sandwich enzyme linked immunosorbentassay(ELISA). RT-PCR was used to detect the expression of glial fibrillary acidic protein(GFAP),brevican and neurocan.Results (1) Compared with the control group, the number of activated astrocytes wereincreased significantly, the cell body become larger, and the increased and extendedprotrusions were woven into mesh in the activation group. The result of ELISA showedthat the content of brevican and neurocan in supernatant gradually increased with the timeprolonging, significant differences compared with the control group (p <0.05). The resultof RT-PCR revealed that the expression of GFAP mRNA, brevican mRNA, and neurocanmRNA also appearred significantly increasing with the extension of time, compared withthe control group with significant difference (p <0.05).(2) After applying Olomoucine intervention in astrocytes, the activated ofastrocytes were suppressed, the number of activated astrocytes were no longer increased,the cell bodies were changed smaller, and the protrusions were reduced and shortened. TheELISA demonstrated that the expression brevican and neurocan with the time prolonging gradually decreased, compared with the activation group with significant difference (p <0.05). The results of RT-PCR indicated that brevican mRNA and neurocan mRNA withthe time prolonging gradually decreased, compared with the activation group withsignificant difference (p <0.05).Conclusions1. The expression of brevican and neurocan were significant increasedafter astrocytes activation;2. Olomoucine could be down-regulated the expression of brevican andneurocan via inhibiting the astrocytes activation.