淫羊藿总黄酮对LPS诱导BV2小胶质细胞炎症反应的作用
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摘要
目的探讨淫羊藿总黄酮(HEP)对脂多糖(LPS)诱导的BV2小胶质细胞炎症反应的作用。方法常规培养BV2小胶质细胞,将其分为对照组、LPS组、20mg/L HEP+LPS组、40mg/L HEP+LPS组、60mg/L HEP+LPS组,后3组先用相应浓度HEP预保护1h,再给予1mg/L的LPS处理细胞6h。采用四甲基偶氮唑蓝(MTT)法检测各组细胞活力,实时反转录聚合酶链反应检测白细胞介素-1β(IL-1β)和肿瘤坏死因子-α(TNF-α)mRNA的表达。结果 1mg/L的LPS和20、40、60mg/L的HEP对BV2小胶质细胞活力没有影响(F=0.59,P>0.05)。LPS可明显上调BV2小胶质细胞炎症因子IL-1β和TNF-αmRNA的表达水平(F=28.94、53.28,q=5.908、20.930,P<0.01);与LPS组相比,20、40、60mg/L的HEP预处理均能明显抑制LPS诱导的IL-1β和TNF-αmRNA的表达(q=5.097～7.669,P<0.01)。结论 HEP可明显抑制LPS诱导的BV2小胶质细胞IL-1β和TNF-αmRNA的表达,提示其可能具有抗炎作用。
Objective To investigate the effect of total flavonoids of Herba Epimedii(HEP)against lipopolysaccharide(LPS)-induced inflammatory response in BV2 microglial cells. Methods BV2 microglial cells were cultured using conventional methods and then divided into control group,LPS group,20 mg/L HEP+LPS group,40 mg/L HEP+LPS group,and 60 mg/L HEP+LPS group.The cells in the latter three groups were pretreated with HEP for 1 hour and then treated with 1 mg/L LPS for6 hours.MTT assay was used to measure cell viability,and real-time RT-PCR was used to measure the mRNA expression of interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α). Results LPS at a concentration of 1 mg/L and HEP at concentrations of 20,40,and 60 mg/L had no influence on the viability of BV2 microglial cells(F=0.59,P>0.05).LPS significantly upregulated the mRNA expression of inflammatory factors IL-1βand TNF-αin BV2 microglial cells(F=28.94 and 53.28,q=5.908 and20.930,P<0.01).Compared with the LPS group,the 20,40,and 60 mg/L HEP+LPS groups had significant reductions in the mRNA expression of IL-1βand TNF-αinduced by LPS(q=5.097-7.669,P<0.01). Conclusion HEP can significantly inhibit the mRNA expression of IL-1βand TNF-αinduced by LPS in BV2 microglial cells,suggesting that HEP may have an anti-inflammatory effect.
Objective To investigate the effect of total flavonoids of Herba Epimedii(HEP)against lipopolysaccharide(LPS)-induced inflammatory response in BV2 microglial cells. Methods BV2 microglial cells were cultured using conventional methods and then divided into control group,LPS group,20 mg/L HEP+LPS group,40 mg/L HEP+LPS group,and 60 mg/L HEP+LPS group.The cells in the latter three groups were pretreated with HEP for 1 hour and then treated with 1 mg/L LPS for6 hours.MTT assay was used to measure cell viability,and real-time RT-PCR was used to measure the mRNA expression of interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α). Results LPS at a concentration of 1 mg/L and HEP at concentrations of 20,40,and 60 mg/L had no influence on the viability of BV2 microglial cells(F=0.59,P>0.05).LPS significantly upregulated the mRNA expression of inflammatory factors IL-1βand TNF-αin BV2 microglial cells(F=28.94 and 53.28,q=5.908 and20.930,P<0.01).Compared with the LPS group,the 20,40,and 60 mg/L HEP+LPS groups had significant reductions in the mRNA expression of IL-1βand TNF-αinduced by LPS(q=5.097-7.669,P<0.01). Conclusion HEP can significantly inhibit the mRNA expression of IL-1βand TNF-αinduced by LPS in BV2 microglial cells,suggesting that HEP may have an anti-inflammatory effect.
引文
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[2]DE WAAL G M,ENGELBRECHT L,DAVIS T,et al.Correlative light-electron microscopy detects lipopolysaccharide and its association with fibrin fibers in Parkinson’s disease,Alzheimer’s disease and type 2diabetes mellitus[J].Scientific Reports,2018,8(1):16798.
[3]GUO Y S,LIANG P Z,LU S Z,et al.ExtracellularαB-crystallin modulates the inflammatory responses[J].Biochemical and Biophysical Research Communications,2019,508(1):282-288.
[4]HENEKA M T,KUMMER M P,LATZ E.Innate immune activation in neurodegenerative disease[J].Nature Reviews Immunology,2014,14(7):463-477.
[5]LIDDELOW S A,GUTTENPLAN K A,LARKE L E,et al.Neurotoxic reactive astrocytes are induced by activated microglia[J].Nature,2017,541(7638):481-487.
[6]CHEN Meixia,WU Jinfeng,LUO Qingli,et al.The anticancer properties of herba epimedii and its main bioactive componentsicariin and icarisideⅡ[J].Nutrients,2016,8(9):563.
[7]SZE S C,TONG Y,NG T B,et al.Herba epimedii:anti-oxidative properties and its medical implications[J].Molecules,2010,15(11):7861-7870.
[8]ZHANG D W,ZHANG J C,FONG C,et al.Herba epimedii flavonoids suppress osteoclastic differentiation and bone resorption by inducing G2/M arrest and apoptosis[J].Biochimie,2012,94(12):2514-2522.
[9]邓炜,郑民强,张静,等.两种黔产淫羊藿总黄酮对痴呆大鼠学习记忆的影响[J].中国药理学通报,2011,27(6):868-871.
[10]XIAO H H,FUNG C Y,MOK S K,et al.Flavonoids from Herba epimedii selectively activate estrogen receptor alpha(ER alpha)and stimulate ER-dependent osteoblastic functions in UMR-106cells[J].Journal of Steroid Biochemistry and Mole-cular Biology,2014,143:141-151.
[11]WU Lin,DU Zhongrui,XU Aili,et al.Neuroprotective effects of total flavonoid fraction of the Epimedium koreanum Nakai extract on dopaminergic neurons:in vivo and in vitro[J].Biomedicine&Pharmacotherapy,2017,91:656-663.
[12]宋来新,张长城,王婷,等.淫羊藿总黄酮通过抑制MAPK/NF-κB信号通路减轻自然衰老大鼠脑组织炎症反应[J].中国药理学通报,2017,33(1):84-90.
[13]冯晓庆,袁良杰,惠雅,等.E2对MPP+诱导SH-SY5Y细胞损伤保护作用及G15的阻断效应[J].青岛大学学报(医学版),2018,54(1):6-9.
[14]张学杰,任晓璠,陈文芳.Rg1对胶质细胞iNOS基因表达的抑制作用及RU486对其影响[J].青岛大学医学院学报,2017,53(2):133-135,139.
[15]WANG Lili,LI Yu,GUO Yubo,et al.Herba epimedii:an ancient Chinese herbal medicine in the prevention and treatment of osteoporosis[J].Current Pharmaceutical Design,2016,22(3):328-349.
[16]CHEN W F,MOK S K,WANG X L,et al.Total flavonoid fraction of the Herba epimedii extract suppresses urinary calcium excretion and improves bone properties in ovariectomised mice[J].British Journal of Nutrition,2011,105(2):180-189.
[17]MOK S K,CHEN W F,LAI W P,et al.Icariin protects against bone loss induced by oestrogen deficiency and activates oestrogen receptor-dependent osteoblastic functions in UMR106cells[J].British Journal of Pharmacology,2010,159(4):939-949.
[18]ZHANG Jinfang,LI Guo,MENG Chunling,et al.Total flavonoids of Herba Epimedii improves osteogenesis and inhibits osteoclastogenesis of human mesenchymal stem cells[J].Phytomedicine,2009,16(6/7):521-529.
[19]LIU Yiheng,ZANG Hongmin,ZHANG Haiying,et al.Effect of herba epimedii flavone on expression of OPG and RANKL in rat osteoblasts[J].Journal of Chinese Medicinal Materiales,2005,28(12):1076-1078.
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[21]WANG Chengcheng,SU Jiayan,CAI Jiyan,et al.Response surface analysis for the optimization of extraction condition for polysaccharides from Epimedium polysaccharides and studies on its tumor immune activities[J].Acta Pharmaceutica Sinica,2016,51(9):1464-1471.
[22]韩贵芳,张长城,陈茜,等.淫羊藿总黄酮通过AMPK/SIRT1/NF-κB信号通路减轻自然衰老大鼠睾丸组织炎症反应[J].天然产物研究与开发,2018,30(9):1489-1493.
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