【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。更多还原

【Abstract】 Objective To observe the inflammatory changes of microglial cells after the intervention of oleic acid(OA) and complement5a(C5a) receptor antagonist(PMX53),explore the role of C5a-C5 aR 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 cells were randomly divided into normal control group,OA 100 μmol/L treatment group,OA 100 μmol/L+PMX53 100 nmol/L treatment group,OA 200 μmol/L treatment group,OA 200 μmol/L+PMX53 100 nmol/L treatment group.The expression changes of Iba-1,C5 aR protein and tumor necrosis factor-α(TNF-α) in microglial cells after intervention of different concentrations of OA and OA+PMX53 were detected 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 C5 aR protein and concentrations of TNF-α were statistically significantly higher than those in control group(P<0.01),the expression of Iba-1 and concentrations of TNF-α in OA+PMX53 intervention group were statistically significantly lower than those in OA group(P <0.01).The expression of C5 aR protein in OA + PMx53 intervention group was statistically significantly lower than that in OA group(OA 100 μmol/L treatment group vs.OA 100 μmol/L+PMX53 100 nmol/L treatment group:P<0.05;OA 200 μmol/L treatment group vs.OA 200 μmol/L+PMX53 100 nmol/L treatment group:P<0.01).The expressions of Iba-1 and C5 aR 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<0.01).Conclusion C5 aR receptor antagonist can suppress inflammatory reaction of microglial cells induced by OA,which indicates that C5a-C5 aR may participate in inflammatory reaction of microglial cells induced by OA.Therefore,C5 aR receptor antagonist may protect the brain tissues from inflammation-induced damage.更多还原