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
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摘要
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.
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.
引文
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[2]RENAULT-MIHARA F,KATOH H,IKEGAMI T.Beneficial compaction of spinal cord lesion by migrating astrocytes through glycogen synthase kinase-3 inhibition[J].EMBO Mol Med,2011,3(11):682-696.
[3]ZHANG L,WU Y,LI Y.Tanshinone IIA improves miR-133expression through MAPK ERK1/2 pathway in hypoxic cardiac myocytes[J].Cell Physiol Biochem,2012,30(4):843-852.
[4]LIU JJ,LIN DJ,LIU P Q.Induction of apoptosis and inhibition of cell adhesive and invasive effects by tanshinone IIA in acute promyelocytic leukemia cells in vitro[J].J Biomed Sci,2006,13(6):813-823.
[5]CHEN G,PARK C K,XIE R G.Connexin-43 induces chemokine release from spinal cord astrocytes to maintain late-phase neuropathic pain in mice[J].Brain,2014,137(Pt 8):2193-2209.
[6]SHI Z,PAN B,FENG S.The emerging role of long non-coding RNA in spinal cord injury[J].J Cell Mol Med,2018,15(3):1231-1234.
[7]LU M,LUO Y,HU P.Tanshinone IIA inhibits AGEs-induced proliferation and migration of cultured vascular smooth muscle cells by suppressing ERK1/2 MAPK signaling[J].Iran J Basic Med Sci,2018,21(1):83-88.
[8]ZHANG W L,CAO Y A,XIA J,et al.Neuroprotective effect of tanshinone IIA weakens spastic cerebral palsy through inflammation,p38MAPK and VEGF in neonatal rats[J].Mol Med Rep,2018,17(1):2012-2018.
[9]HUANG S T,HUANG C C,HUANG W L,et al.Tanshinone IIA induces intrinsic apoptosis in osteosarcoma cells both in vivo and in vitro associated with mitochondrial dysfunction[J].Sci Rep,2017,7:40382.
[10]LU B L,LI J,ZHOU J,et al.Tanshinone IIA decreases the levels of inflammation induced by Abeta1-42 in brain tissues of Alzheimer′s disease model rats[J].Neuroreport,2016,27(12):883-893.