Department of Ophthalmology, The First Affiliated Hospital of Zhengzhou University, East Jianshe Road, Zhengzhou, 450001, People's Republic of China.
J Mol Neurosci. 2019 May;68(1):111-119. doi: 10.1007/s12031-019-01289-w. Epub 2019 Mar 9.
Studies have shown that papaverine can inhibit lipopolysaccharide (LPS)-induced microglial activation. The retinal primary microglia of newborn SD rats were isolated and purified, and a LPS-induced microglia activation model was established. The protein phosphorylation level of the signaling pathway was detected by western blotting. The transcription and expression of TNF-α, IL-1β, and IL-10 were respectively detected by RT-PCR and ELISA to observe the abnormal activation of primary microglia. The cAMP inhibitor Rp-isomer, PKA inhibitor H89, and MEK inhibitor U0126 were separately added to further investigate the role of MEK/Erk in PAP inhibition of primary microglial activation and the relationship between cAMP/PKA and MEK/Erk. It was found that the level of MEK phosphorylation was upregulated after LPS stimulation, which was blocked by 10 μg/ml of papaverine.10μM U0126 significantly inhibited TNF-α and IL-1β and increased IL-10 transcription and expression in retinal microglia (P < 0.01). Both Rp-isomer and H89 upregulated the phosphorylation levels of MEK and Erk. Papaverine may inhibit inflammatory factors and promote the expression of anti-inflammatory factors through the cAMP/PKA and MEK/Erk pathway, thereby inhibiting LPS-induced activation of primary retinal microglia, and the MEK/Erk pathway may be partially regulated by cAMP/PKA, which can provide theoretical basis and experimental basis for its protection of the central nervous system.
研究表明罂粟碱可以抑制脂多糖(LPS)诱导的小胶质细胞激活。分离并纯化新生 SD 大鼠的视网膜原代小胶质细胞,并建立 LPS 诱导的小胶质细胞激活模型。通过 Western blot 检测信号通路的蛋白磷酸化水平。通过 RT-PCR 和 ELISA 分别检测 TNF-α、IL-1β 和 IL-10 的转录和表达,观察原代小胶质细胞的异常激活。分别添加 cAMP 抑制剂 Rp-异构体、PKA 抑制剂 H89 和 MEK 抑制剂 U0126,进一步研究 MEK/Erk 在 PAP 抑制原代小胶质细胞激活中的作用以及 cAMP/PKA 和 MEK/Erk 之间的关系。结果发现,LPS 刺激后 MEK 磷酸化水平上调,被 10μg/ml 罂粟碱阻断。10μM U0126 显著抑制 TNF-α 和 IL-1β,增加视网膜小胶质细胞中 IL-10 的转录和表达(P<0.01)。Rp-异构体和 H89 均上调 MEK 和 Erk 的磷酸化水平。罂粟碱可能通过 cAMP/PKA 和 MEK/Erk 通路抑制炎症因子,促进抗炎因子的表达,从而抑制 LPS 诱导的原代视网膜小胶质细胞激活,而 MEK/Erk 通路可能部分受 cAMP/PKA 调节,为其对中枢神经系统的保护作用提供了理论依据和实验依据。