布洛芬对少突神经胶质细胞生长发育的影响

摘要： 目的 探究布洛芬对少突胶质细胞 （OLs） 生长发育的影响。方法 健康清洁级SD成年大鼠6只， 提取 OLs， 扩增后加入溶血磷脂酸 （LPA） 观察细胞形态变化。使用出生24~48 h的新生鼠6只， 提取少突胶质细胞前体细胞 （OPCs）， 将OPCs接种后随机分为对照组、 LPA组、 布洛芬组、 LPA+布洛芬组。贴壁3 d后观察细胞形态， 对照组加入生理盐水， 其他3组分别加入LPA （1.0 μmol/L）、 布洛芬盐水溶液 （100 μmol/L） 及两者的混合液。药物持续干预7 d 后观察细胞形态， 分别对OPCs特异性免疫标志物血小板源生长因子受体α （PDGFR-α） 和OLs特异性免疫标志物髓鞘碱性蛋白 （MBP） 行免疫荧光染色观察OPCs及OLs形态并细胞计数。使用Western blot法比较各组MBP相对表达量。结果 LPA作用于成熟OLs后细胞突起减少。OPCs贴壁3 d时各组细胞发育良好。药物干预7 d后， 布洛芬组和LPA+布洛芬组中细胞突起明显多于对照组和LPA组。布洛芬组、 LPA+布洛芬组的MBP阳性细胞计数多于对照组和LPA组 （P＜0.01）。Western blot结果显示， LPA组的MBP表达量明显少于其他3组 （P＜0.01）， 布洛芬组表达量明显多于LPA＋布洛芬组 （P＜0.01）。结论 LPA对OPCs的生长发育有毒性作用， 对成熟OLs的正常生长具有抑制作用， 一定浓度的布洛芬能够显著抑制LPA对OPCs及OLs的细胞毒性作用。

Effects of ibuprofen on the growth and development of oligodendrocytes

Abstract: Objective To study the effects of ibuprofen on the growth and development of oligodendrocytes. Methods A total of 6 clean and healthy adult female SD (Sprague Dawley) rats were used for extracting and culturing of oligodendrocytes (OLs). Lysophosphatidic acid (LPA) was then added, and the morphological changes of OLs pre-treatment and post-treatment were observed. Then 6 newborn rats (born 24-48 h) were used for mixed glial cell extraction from the cortex, then the OPCs were inoculated into the culture plates and randomly divided into control group, ibuprofen group, lysophosphatidic acid (LPA) group and LPA+ ibuprofen group. After the adhering of the cells in each group for three days, cell morphology was observed, and the drugs were added as interventions. The control group was treated with normal saline, and the other 3 groups were added with saline solution of ibuprofen (100 μmol/L), LPA (1.0 μmol/L) and the mixture of them. The cell morphological changes were observed after 7-day intervention. The morphology of OPCs and OLs were observed by immunofluorescence staining through OPCs’specific immune markers (platelet-derived growth factor receptor alpha, PDGFR-α) and OLs’specific immune markers (myelin basic protein, MBP) along with cell count of mature OLs. Western blot assay was used to detect the relative expression level of MBP in each group. Results After the treatment with LPA to the mature OLs, protrusions were shrinking and became very sparse. The morphology of cells developed well in each group after cell adhering for 3 days. After drug intervention for 7 days, more cell protrusions and branches were observed in ibuprofen group and LPA+ ibuprofen group than those of the control group and LPA group. The results of cell count showed that the number of MBP positive cells was significantly higher in the ibuprofen group and LPA+ibuprofen group than that in the control group and LPA group (P＜0.01). The results of Western blot assay showed that the MBP protein expression was significantly less in LPA group than the other three groups (P＜0.01), and the expression was significantly higher in the ibuprofen group than that of LPA + ibuprofen group (P＜0.01). Conclusion LPA has a toxic effect on the growth and development of OPCs, and it has an inhibitory effect on the normal growth of mature OLs. A certain concentration of ibuprofen can significantly inhibit the cytotoxicity of LPA on OPCs and OLs, and promote the formation and maintenance of mature OLs.