玉女煎对脂多糖诱导的BV2细胞炎性损伤的保护作用及机制研究
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摘要
目的:研究玉女煎对脂多糖(LPS)诱导的BV2细胞的保护作用并探讨其可能的作用机制。方法:利用LPS诱导BV2细胞炎性损伤模型,玉女煎处理细胞24 h,MTT法检测细胞凋亡,Griess法检测NO分泌,ELISA法检测IL-1β、IL-6及TNF-α分泌水平,Real-time PCR和Western blot分别检测iNOS和COX-2 mRNA及蛋白表达的变化。结果:与空白对照组相比,LPS刺激能显著降低BV2细胞存活率,诱导NO、IL-1β、IL-6及TNF-α的分泌,增加iNOS、COX-2 mRNA及蛋白的表达,促进NF-κB p65由细胞质进入细胞核。与LPS组相比,1、2 mg/mL玉女煎预处理BV2后,细胞存活率显著升高,NO、IL-1β及TNF-α分泌量显著下降;0.1、1、2 mg/mL玉女煎预处理可显著降低IL-6分泌、抑制iNOS、COX-2 mRNA及蛋白表达;2 mg/mL玉女煎预处理能明显抑制NF-κB p65由细胞质进入细胞核。结论:玉女煎可能通过抑制NF-κB活性,进而抑制炎症因子分泌及相关酶表达从而发挥抗LPS诱导的BV2细胞炎症作用。
Objective: To study the protective effect and the mechanism of Yunvjian on lipopolysaccharide( LPS) induced inflammation in BV2 cells. Methods: BV2 cells Inflammation was induced by LPS,and cells were treated with different concentrations of Yunvjian for 24 h. The cell viability was determined by MTT assay. The NO production was detected by Griess reaction method,the secretions of IL-1β、IL-6 and TNF-α were determined by ELISA assay. The levels of mRNA and protein were determined by real-time PCR and western blot. Results:Compared with the control group,LPS significantly reduced the survival rate of RAW264. 7 cells,increased the secretions of NO,IL-1β and TNF-α. It also increased the expressions of iN OS and COX-2 mRNA and protein,and promoted NF.-κB p65 to enter the nucleus from the cytoplasm. Compared with LPS group,cell viabilities of 1 and 2 mg/mL Yunvjian pretreatment were significantly increased,and the secretions of NO,IL-1β and TNF-α were significantly decreased. 0. 1,1 and 2 mg/mL Yunvjian pretreatment significantly reduced the secretion of IL-6,inhibited the expressions of iN OS,COX-2 mRNA and protein. 2 mg/mL Yunvjian pretreatment inhibited NF-κB p65 from entering into nucleus. Conclusion: Yunvjian can inhibit LPS-induced inflammation in BV2 cells,possibly through inhibiting NF-κB activity,thereby inhibiting inflammatory factor secretion and related enzyme expression.
Objective: To study the protective effect and the mechanism of Yunvjian on lipopolysaccharide( LPS) induced inflammation in BV2 cells. Methods: BV2 cells Inflammation was induced by LPS,and cells were treated with different concentrations of Yunvjian for 24 h. The cell viability was determined by MTT assay. The NO production was detected by Griess reaction method,the secretions of IL-1β、IL-6 and TNF-α were determined by ELISA assay. The levels of mRNA and protein were determined by real-time PCR and western blot. Results:Compared with the control group,LPS significantly reduced the survival rate of RAW264. 7 cells,increased the secretions of NO,IL-1β and TNF-α. It also increased the expressions of iN OS and COX-2 mRNA and protein,and promoted NF.-κB p65 to enter the nucleus from the cytoplasm. Compared with LPS group,cell viabilities of 1 and 2 mg/mL Yunvjian pretreatment were significantly increased,and the secretions of NO,IL-1β and TNF-α were significantly decreased. 0. 1,1 and 2 mg/mL Yunvjian pretreatment significantly reduced the secretion of IL-6,inhibited the expressions of iN OS,COX-2 mRNA and protein. 2 mg/mL Yunvjian pretreatment inhibited NF-κB p65 from entering into nucleus. Conclusion: Yunvjian can inhibit LPS-induced inflammation in BV2 cells,possibly through inhibiting NF-κB activity,thereby inhibiting inflammatory factor secretion and related enzyme expression.
引文
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[4]Vizuete M L, Merino M, Venero J L, et al. Histamine infusion induces a selective dopaminergic neuronal death along with an inflammatory reaction in rat substantia nigra. J Neurochem, 2000, 75(2)∶540～552.
[5]Hu Y, Zeng Z, Wang B, et al. Trans-caryophyllene inhibits amyloid β (Aβ) oligomer-induced neuroinflammation in BV-2 microglial cells. Int Immunopharmacol, 2017, 51(1)∶91～98.
[6]Tsai C F, Kuo Y H, Yeh W L, et al. Regulatory effects of caffeic acid phenethyl ester on neuroinflammation in microglial cells. Int J Mol Sci, 2015, 16(3)∶5572～5589.
[7]雷莉妍,唐志书,刘妍如, 等. 玉女煎HPLC指纹图谱的研究. 中药材, 2016, 39(1)∶113～116.
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[10]王晓晨, 吉爱国. NF-κB信号通路与炎症反应. 生理科学进展, 2014; 45(1)∶68～71.
[11]Sang W, Zhong Z, Linghu K, et al. Siegesbeckia pubescens Makino inhibits Pam3CSK4-induced inflammation in RAW 264.7 macrophages through suppressing TLR1/TLR2-mediated NF-κB activation. Chin Med, 2018, 13(1)∶37.
[12]Jouda J B, Mfotie Njoya E, Mbazoa C D, et al. Lambertellin from Pycnoporus sanguineus MUCL 51321 and its anti-inflammatory effect via modulation of MAPK and NF-κB signaling pathways. Bioorg Chem, 2018, 80(1)∶216～222.
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