DHEA和EPEA对小神经胶质细胞和星形胶质细胞的体外作用研究

Effects of DHEA and EPEA on Microglia and Astrocytes In Vitro

目的探究二十二碳六烯酸单乙醇酰胺(DHEA)和二十碳五烯酸单乙醇酰胺(EPEA)在脊柱外伤后对小神经胶质细胞和星形胶质细胞的作用,旨在为中枢神经性疼痛治疗提供理论依据和实验基础。方法对DHEA和EPEA采用同时给药和延迟给药两种方式,同时治疗分为control组(无血清培养液)、100 ng/ml LPS组(100 ng/ml LPS),DHEA组(8μmol/L DHEA及LPS),EPEA组(8μmol/L EPEA及LPS),延迟治疗Delay control组加入无血清培养液,Delay LPS组、Delay DHEA组、Delay EPEA组分别加入100 ng/ml LPS进行激活,小神经胶质细胞激活16 h、星形胶质细胞激活24 h后加入无血清培养液到Delay LPS组,分别加入8μmol/L DHEA及8μmol/L EPEA到Delay DHEA组和Delay EPEA组,观察小神经胶质细胞形态变化、突起百分比、iNOS及GFAP表达。结果与control组相比,LPS处理小神经胶质细胞和星形胶质细胞后细胞胞体变大,突起缩短(P<0.001),iNOS表达显著增加[(3.34±1.06)%vs(67.79±4.45)%,P<0.001],GFAP表达显著升高[(21.36±5.17)%vs(40.12±6.39%),P<0.001];经EPEA和DHEA同时、延期给药处理后细胞突起恢复(P<0.001),iNOS表达显著降低[(35.57±5.34)%,P<0.001],GFAP表达也显著下降[(23.12±4.26)%,P<0.001]。结论DHEA和EPEA在同时、延期给药两种处理下均能改变激活态小神经胶质细胞和星形胶质细胞的形态和状态。

Objective To investigate the effects of docosahexaenoyl ethanolamide(DHEA)and eicosapentaenoyl ethanolamide(EPEA)on microglia and astrocytes after spinal trauma,aiming to provide theoretical and experimental basis for the treatment of central nervous pain.Methods DHEA and EPEA were treated with simultaneous administration and delayed administration,and were divided into control group(serum-free culture medium),100 ng/ml LPS group(100 ng/ml LPS),and DHEA group(8μmol/L DHEA and LPS).EPEA group(8μmol/L EPEA and LPS),Delay control group were added with serum-free culture medium,Delay LPS group,Delay DHEA group and Delay EPEA group were activated with 100 ng/ml LPS,respectively.After microglial cells were activated for 16 h and astroglial cells were activated for 24 h,serum-free medium was added to Delay LPS group,8μmol/L DHEA and 8μmol/L EPEA were added to Delay DHEA group and Delay EPEA group,respectively.The morphological changes of microglial cells,the percentage of protrusions,the expression of iNOS and GFAP were observed.Results Compared with the control group,the cell bodies of microglia and astrocytes treated with LPS became larger,the processes were shortened(P<0.001),the expression of iNOS was significantly increased[(3.34±1.06)%vs(67.79±4.45)%,P<0.001],and the expression of GFAP was significantly increased[(21.36±5.17)%vs(40.12±6.39)%,P<0.001].After treatment with EPEA and DHEA at the same time and delayed administration,the cell protrusions recovered(P<0.001),the expression of iNOS decreased significantly[(35.57±5.34)%,P<0.001],and the expression of GFAP also decreased significantly[(23.12±4.26)%,P<0.001].Conclusion Both DHEA and EPEA can change the morphology and state of activated microglia and astrocytes under simultaneous and delayed administration.