摘要
目的观察油酸(oleic acid,OA)及C5a受体(C5aR)拮抗剂(PMX53)干预后小胶质细胞的炎性改变,探讨C5a-C5aR在油酸诱导的小胶质细胞炎症中的作用。方法取新生1 d龄小鼠,分离脑组织,原代培养小胶质细胞并进行分离纯化及鉴定。纯化小胶质细胞随机分成5组,分别为正常对照组、OA 100μmol/L处理组、OA 100μmol/L+PMX53 100 nmol/L处理组、OA 200μmol/L处理组、OA 200μmol/L+PMX53 100 nmol/L处理组。应用Western印迹、ELISA法检测不同浓度OA及OA+PMX53干预后小胶质细胞Iba-1、C5aR蛋白及TNF-α表达变化。结果成功培养小胶质细胞,且纯度≥99%。不同浓度OA干预后,Iba-1、C5aR蛋白表达及TNF-α浓度较对照组明显升高(P<0.01);OA+PMX53干预组较OA组Iba-1及TNF-α浓度有所下降(P<0.01);OA+PMx53干预组较OA组C5aR蛋白表达有所下降(OA100 vs.OA100+PMX53,P<0.05,OA200 vs.OA200+PMX53,P<0.01)。200μmol/L OA干预组较100μmol/L OA干预组Iba-1、C5aR蛋白表达及TNF-α浓度高(P<0.01)。结论应用C5aR拮抗剂可以抑制油酸诱导的小胶质细胞的炎症反应,提示C5a-C5aR参与了油酸诱发的炎症反应,推测C5aR拮抗剂在神经系统炎性损伤中具有神经保护作用。
Objedive To observe the inflammatory changes of microglial cells after the intervention of oleic acid （OA） and complement 5a （C5a） receptor antagonist （PMX53）, explore the role of C5a-CSaR in inflammation of microglial cells induced by OA. Methods The brain tissue samples were isolated from neonatal one-day-old mice, primary microglial cells were isolated, purified, and identified. Purified microglial ceils were randomly divided into normal control group, OA 100 μmnol/L treatment group, OA 100 μmol/L＋PMX53 100 nmol/L treat- ment group, OA 200 μmol/L treatment group, OA 200 μmol/L＋PMX53 100 nmol/L treatment group. The expression changes of Iba-1, CSaR protein and tumor necrosis factor-or （TNF-α） in microglial cells after intervention of different concentrations of OA and OA＋PMX53 were de- tected by Western blot and ELISA. Results Primary microglial cells were successfully cultured, and the puritification was no less than 99%. After intervention of different concentrations of OA, the expressions of Iba-1 and C5aR protein and concentrations of TNF-α were statistically significantly higher than those in control group （P〈0. O1 ）, the expression of Iba-1 and concentrations of TNF-a in OA＋PMX53 intervention group were statistically significantly lower than those in OA group （P〈0. O1 ） . The expression of C5aR protein in OA＋PMx53 intervention group was statistically significantly lower than that in OA group （ OA 100 μmol/L treatment group vs. OA 100 txmol/L＋PMX53 100 nmol/L treatment group： P〈0. 05; OA 200 μmol/L treatment group vs. OA 200 txmol/L＋PMX53 100 nmol/L treatment group： P〈0. O1 ） . The ex- pressions of Iba-1 and C5aR protein and concentrations of TNF-α in OA 200 μmol/L treatment group were statistically significantly higher than those in OA 100 μmol/L treatment group （P〈O. 01 ） . Conclusion C5aR receptor antagonist can suppress inflammatory reaction of micrnglial cells induced by OA, which indicates that C5a-CSaR n-my participate in inflamm
作者
刘艳
龙文君
卢慧玲
刘桐林
LIU Yan LONG Wen-Jun LU Hui-Ling et al(Department of Pediatrics, Tongfi Hospital Affiliated to Tong]i Medical College of Huazhong University of Science and Tech- nology, Wuhan, Hubei 430030, Chin)
出处
《中国妇幼保健》
CAS
2017年第20期5113-5116,共4页
Maternal and Child Health Care of China
基金
国家自然科学基金资助项目(81401277)
关键词
油酸
C5A受体
小胶质细胞
炎症
Oleic acid
Complement 5a receptor
Microglial cell
Inflammation
