丹参酮ⅡA对大鼠脊髓损伤后星形胶质细胞增殖的MAPK通路的影响

Effects of tanshinone ⅡA on MAPK pathway of astrocyte proliferation after spinal cord injury in rats

目的探讨丹参酮ⅡA(tanshinoneⅡA,TⅡA)对大鼠脊髓损伤(spinal cord injury,SCI)后星形胶质细胞增殖的MAPK通路的影响。方法建立大鼠SCI模型,获得星形胶质细胞,培养14 d后,采用免疫荧光法检测星形胶质细胞中胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)的表达;MTT法选择TⅡA抑制星形胶质细胞生长的最适浓度和时间;Western blot法检测MAPK通路各蛋白的磷酸化情况。结果星形胶质细胞中有GFAP特异性表达;TⅡA抑制星形胶质细胞生长的最适浓度为15μg/mL,最适作用时间为36 h;15μg/m L TⅡA作用0和4 h,星形胶质细胞中p-JNK高于8和12 h,ERK1/2和p38无变化。结论 TⅡA主要通过JNK依赖性途径对星形胶质细胞的生长进行调控。

Objective To investigate the effect of tanshinoneⅡA(TⅡA)on the MAPK pathway of astrocyte proliferation after spinal cord injury(SCI)in rats. Methods Rat model of SCI was established. The obtained astrocytes were cultured for 14 d and determined for the expression of glial fibrillary acidic protein(GFAP)by immunofluorescence assay(IFA).The concentration and time of TⅡA for inhibiting the growth of astrocytes were optimized by MTT assay. The phosphorylation of various proteins in MAPK pathway was determined by Western blot. Results GFAP was expressed specifically in astrocytes. The optimal concentration and time of TⅡA for inhibiting the growth of astrocytes were 15 g/m L and 36 h respectively. The p-JNK levels in castrocytes after treatment with 15 μg/m L TⅡA for 0 and 4 h were higher than those after treatment for 8 and 12 h,while those of ERK1/2 and p38 showed no change. Conclusion TⅡA mainly regulated the growth of astrocytes by JNK dependent pathway.