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目的研究替米沙坦改善脂多糖(lipopolysaccharide,LPS)诱导的小胶质细胞炎症反应是否与其激活过氧化物酶体增殖物激活受体γ(peroxisome proliferator-activated receptorγ,PPARγ)进而促进小胶质细胞激活表型转化有关。方法采用荧光素酶报告基因检测方法研究替米沙坦对PPARγ的激动活性;建立LPS诱导的小鼠BV-2小胶质细胞炎症模型,在该模型上用0.1~10 mol/L替米沙坦单独处理BV-2细胞,或者1μmol/L替米沙坦与10μmol/L PPARγ特异性阻断剂GW9662共同处理BV-2细胞,用ELISA法检测细胞上清中肿瘤坏死因子-α(tumor necrosis factor-alpha,TNF-α)的含量,用实时定量PCR法检测BV-2小胶质细胞M1型细胞标志物(CD16、CD11b、iNOS)和M2型细胞标记物IL-10的mRNA表达水平变化。结果与溶剂对照组相比,0.1~10 mol/L替米沙坦能浓度依赖地激活PPARγ,对荧光素酶的诱导倍数分别为1.29、1.36和1.45(均P<0.01);在LPS诱导的BV-2小胶质细胞炎症模型上,与溶剂对照组相比,LPS处理组细胞上清中TNF-α的含量显著升高(P<0.01),小胶质细胞M1型标记物CD16、CD11b和iNOS mRNA表达水平显著上调(P<0.05);与LPS处理组相比,0.1~10 mol/L替米沙坦能浓度依赖地降低小胶质细胞上清中TNF-α的含量(P<0.05),10 mol/L替米沙坦能显著下调小胶质细胞M1型标记物CD16、CD11b和iNOS的mRNA表达水平(P<0.01和P<0.05),且0.1 mol/L替米沙坦能显著上调小胶质细胞M2型标记物IL-10的mRNA表达水平(P<0.05);与1 mol/L替米沙坦处理组相比,PPARγ特异性阻断剂GW9662能显著上调CD16的mRNA表达水平(P<0.05)。结论替米沙坦显著抑制LPS诱导的小胶质细胞激活后TNF-α的释放,其作用机制可能与替米沙坦激活PPARγ并促进小胶质细胞M1/M2激活表型的转化密切相关。
Objective To study whether telmisartan improves the microglial inflammatory response induced by lipopolysaccharide (LPS) in combination with its activation of peroxisome proliferator-activated receptor γ (PPARγ) Cell activation phenotypic transformation. Methods Activation of PPARγ by telmisartan was detected by luciferase reporter assay. A mouse model of BV-2 microglial inflammation induced by LPS was established. The model was treated with 0.1-10 mol / L telmisartan BV-2 cells were treated with BV alone or 1μmol / L telmisartan and GW9662 (10μmol / L PPARγ specific inhibitor). ELISA was used to detect the expression of tumor necrosis factor-α (TNF-α, TNF-α, TNF-α and TNF-α). The mRNA expression levels of Ml-2 microglial cell marker (CD16, CD11b, iNOS) and M2 cell marker IL- . Results Compared with the solvent control group, 0.1-100 mol / L telmisartan could activate PPARγ in a concentration-dependent manner, and the induction times of luciferase were 1.29,1.36 and 1.45 (all P <0.01) BV-2 microglial inflammation model, compared with the solvent control group, the content of TNF-α in the supernatant of the LPS-treated group was significantly increased (P <0.01), and microglial M1 markers CD16, CD11b (P <0.05). Compared with LPS treatment group, 0.1-10 mol / L telmisartan could decrease the concentration of TNF-α in the microglia supernatant in a concentration-dependent manner (P < 0.05). Telmisartan at 10 mol / L significantly decreased the mRNA expression of M1, CD16, CD11b and iNOS in microglia (P <0.01 and P <0.05), and 0.1 mol / L telmisartan (P <0.05). Compared with 1 mol / L telmisartan treatment group, GW9662, a specific inhibitor of PPARγ, could up-regulate the expression of CD16 mRNA expression level (P <0.05). Conclusion Telmisartan can significantly inhibit the release of TNF-α after LPS-induced microglial activation, which may be related to the activation of PPARγ by telmisartan and the activation of M1 / M2-activated microglia.