炎宁糖浆抗炎活性及其作用机制研究
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摘要
目的考察炎宁糖浆总提取物及其3种单味药的抗炎活性,并对其作用机制进行初步探索,为临床应用提供科学依据。方法采取二甲苯致小鼠耳肿胀法和新鲜鸡蛋清致大鼠足肿胀法,观察体内抗炎作用;采用MTT法检测其对THP-1细胞活力的影响;ELISA和Western blot检测其对LPS诱导THP-1细胞释放TNF-α等炎性因子及调控其产生的关键蛋白的影响,以探讨总提取物及其单味药可能的抗炎机制。结果体内实验中,炎宁糖浆高、中剂量组均表现出明显的抗炎作用;体外实验中,MTT实验表明鹿茸草和鸭跖草无细胞毒作用,而总提取物及白花蛇舌草分别在125、5 mg·L-1表现出细胞毒活性。ELISA实验表明,总提取物、鹿茸草和鸭跖草均具有抑制TNF-α等炎症因子产生的作用。此外,总提取物还表现出抑制I-κBα磷酸化、NF-κB p65核转位,以及i NOS和COX-2蛋白过表达的作用。结论炎宁糖浆具有较好的抗炎作用,其主要抗炎单味药为鹿茸草和鸭跖草。
Aim To investigate the anti-inflammatory activity of Yanning syrup and its three kinds of herbs, and to explore the mechanism of action, in order to provide scientific basis for its clinical application. Methods The anti-inflammatory effect in vivo was observed by the ear swelling of mice induced by xylene and different hind paw edema mice induced by fresh egg white. MTT assay was applied to determine THP-1 cell viability in cells in in vitro experiment. The enzyme-linked immunosorbent assay( ELISA) and Western blot were used to measure the content of inflammatory cytokines, such as TNF-α, induced by LPS in THP-1 cell, as well as the key proteins which verified the anti-inflammatory activity and mechanism of Yanning syrup and its herbs. Results Yanning syrup high, medium dose groups had significant anti-inflammatory effect in in vivo experiment. The results of MTT indicated that Commelina communis and Monochasma sheareri Maxim. in Yanning syrup had no cytotoxic effect, while the total extract of Yanning syrup and the extract of Hedyotis diffusa Willd. displayed cytotoxicity at 125 mg·L~(-1) and 5 mg·L~(-1). The results of ELISA showed that the total extract of Yanning syrup, Commelina communis and Monochasma sheareri Maxim. could inhibit the production of inflammatory cytokines, such as TNF-α and so on. In addition, the total extract of Yanning syrup also showed inhibitory effects of I-κBα phosphorylation, NF-κB p65 nuclear transfer and iNOS and COX-2 proteins high expression. Conclusions Yanning syrup has a good anti-inflammatory effect, while its main anti-inflammatory herbs are Commelina communis and Monochasma sheareri Maxim..
Aim To investigate the anti-inflammatory activity of Yanning syrup and its three kinds of herbs, and to explore the mechanism of action, in order to provide scientific basis for its clinical application. Methods The anti-inflammatory effect in vivo was observed by the ear swelling of mice induced by xylene and different hind paw edema mice induced by fresh egg white. MTT assay was applied to determine THP-1 cell viability in cells in in vitro experiment. The enzyme-linked immunosorbent assay( ELISA) and Western blot were used to measure the content of inflammatory cytokines, such as TNF-α, induced by LPS in THP-1 cell, as well as the key proteins which verified the anti-inflammatory activity and mechanism of Yanning syrup and its herbs. Results Yanning syrup high, medium dose groups had significant anti-inflammatory effect in in vivo experiment. The results of MTT indicated that Commelina communis and Monochasma sheareri Maxim. in Yanning syrup had no cytotoxic effect, while the total extract of Yanning syrup and the extract of Hedyotis diffusa Willd. displayed cytotoxicity at 125 mg·L~(-1) and 5 mg·L~(-1). The results of ELISA showed that the total extract of Yanning syrup, Commelina communis and Monochasma sheareri Maxim. could inhibit the production of inflammatory cytokines, such as TNF-α and so on. In addition, the total extract of Yanning syrup also showed inhibitory effects of I-κBα phosphorylation, NF-κB p65 nuclear transfer and iNOS and COX-2 proteins high expression. Conclusions Yanning syrup has a good anti-inflammatory effect, while its main anti-inflammatory herbs are Commelina communis and Monochasma sheareri Maxim..
引文
[1] 朱莹莹, 李 敏, 陈 重,等. 炎宁颗粒的化学成分研究[J]. 中成药, 2013, 35(5):988-94.[1] Zhu Y Y, Li M, Chen Z, et al. Chemical constituents from Yanning Granules[J]. Chin Tradit Patent Med, 2013, 35(5):988-94.
[2] 马 静, 许琼明, 吴文倩,等. 炎宁颗粒有效部位化学成分研究[J]. 中成药, 2012, 34(10):1946-8.[2] Ma J, Xu Q M, Wu W Q, et al. Chemical constituents of active fractions of Yanning Granules[J]. Chin Tradit Patent Med, 2012, 34(10):1946-8.
[3] 乡世健, 潘林燕, 王婉菱,等. 鹿茸草抗炎、止咳作用的初步研究[J]. 时珍国医国药, 2012, 23(11):2727-8.[3] Xiang S J, Pan L Y, Wang W L, et al. Preliminary study on anti-inflammatory and antitussive effects of Monochasma sheareri[J]. Lishizhen Med Mater Med Res, 2012, 23(11):2727-8.
[4] 郑 巍, 谭兴起, 郭良君,等. 鹿茸草的化学成分[J]. 中国天然药物, 2012, 10(2):102-4.[4] Zheng W, Tan X Q, Guo L J, et al. Chemical constituents from Monochasma savatieri[J]. Chin J Nat Med, 2012, 10(2):102-4.
[5] 余 昕, 朱 烨, 欧丽兰,等. 鸭跖草抗炎活性部位筛选及抗炎机制[J]. 中成药, 2015, 37(8):1824-7.[5] Yu X, Zhu Y, Ou L L, et al. Screening of anti-inflammatory active sites and anti-inflammatory mechanism of Commelina communis[J]. Chin Tradit Patent Med, 2015, 37(8):1824-7.
[6] 袁红娥, 周兴栋, 孟令杰,等. 鸭跖草的化学成分研究[J]. 中国中药杂志, 2013, 38(19):3304-8.[6] Yuan H E, Zhou X D, Meng L J,et al. Chemical constituents from Commelina communis[J]. Chin J Chin Mater Med, 2013, 38(19):3304-8.
[7] 纪宝玉, 范崇庆, 裴莉昕,等. 白花蛇舌草的化学成分及药理作用研究进展[J]. 中国实验方剂学, 2014, 20(19):235-40.[7] Ji B Y, Fan C Q, Pei L X, et al. Advance on the chemical and pharmacological effects studies of Hedyotis diffusa[J]. Chin J Exp Tradit Med Form, 2014, 20(19):235-40.
[8] 杨 娴, 刘汝青, 杜德荣,等. 白花蛇舌草总黄酮对黑色素瘤A375和B16细胞增殖和凋亡的影响[J]. 皮肤性病诊疗学杂志, 2011, 18(6):359-62.[8] Yang X, Liu R Q, Du D R, et al. Effects of FOD on the proliferation and apoptosis of melanoma A375 and B16 Cells[J]. J Diagn Ther Dermato-Venereol, 2011, 18(6):359-62.
[9] Wang J H, Shu L H, Yang L L, et al. 2-Hydroxy-3-methylanthraquinone from Hedyotis diffusa WILLD induces apoptosis via alteration of Fas/FasL and activation of caspase-8 in human leukemic THP-1 cells[J]. Arch Med Res, 2011, 42(7):577-83.
[10] 刘 征, 吴洪涛, 倪亚光,等. Wnt5a与炎症信号在炎症性疾病中的交叉互动[J]. 生理学报, 2015, 67(4):437-45.[10] Liu Z, Wu H T, Ni Y G, et al. Crosstalk between Wnt5a and inflammatory signaling in inflammation[J]. Acta Physiol Sin, 2015, 67(4):437-45.
[11] 潘 磊, 金 旺, 潘宏宇,等. 魔芋葡甘露聚糖(KGM)对LPS诱导的THP-1源巨噬细胞产生细胞因子的影响[J]. 中华中医药学刊, 2017, 35(6):1496-501.[11] Pan L, Jin W, Pan H Y, et al. Effect of Konjac Glucomannan ( KGM) on LPS-induced THP-1 derived macrophages produce cytokines[J]. Chin Arch Tradit Chin Med, 2017, 35(6):1496-501.
[12] 袁 琴, 袁 丁, 周志勇,等. 竹节参齐墩果烷皂苷对RAW264.7巨噬细胞SIRT1活性影响及抗炎作用研究[J]. 中国药理学通报, 2016, 32(3):349-54.[12] Yuan Q, Yuan D, Zhou Z Y, et al. Anti-inflammatory effect of Chikusetsu oleanane saponin on RAW264.7 cell through regulating SIRT1 activity[J]. Chin Pharmacol Bull, 2016, 32(3):349-54.
[13] 王晓晨, 吉爱国. NF-κB信号通路与炎症反应[J]. 生理科学进展, 2014, 45(1):68-71.[13] Wang X C, Ji A G. NF-κB signaling pathway and inflammatory response[J]. Prog Physiol Sci, 2014, 45(1):68-71.
[14] 袁 昊, 曾 晖, 肖德明,等. 白藜芦醇通过NF-κB信号通路抑制软骨细胞炎症因子的表达[J]. 中华骨与关节外科杂志, 2016, 9(1):75-9.[14] Yuan H, Zeng H, Xiao D M, et al. Resveratrol inhibits the expression of inflammatory cytokines in cartilage cells through NF-κB signaling pathway[J]. Chin J Bone Joint Surg, 2016, 9(1):75-9.
[2] 马 静, 许琼明, 吴文倩,等. 炎宁颗粒有效部位化学成分研究[J]. 中成药, 2012, 34(10):1946-8.[2] Ma J, Xu Q M, Wu W Q, et al. Chemical constituents of active fractions of Yanning Granules[J]. Chin Tradit Patent Med, 2012, 34(10):1946-8.
[3] 乡世健, 潘林燕, 王婉菱,等. 鹿茸草抗炎、止咳作用的初步研究[J]. 时珍国医国药, 2012, 23(11):2727-8.[3] Xiang S J, Pan L Y, Wang W L, et al. Preliminary study on anti-inflammatory and antitussive effects of Monochasma sheareri[J]. Lishizhen Med Mater Med Res, 2012, 23(11):2727-8.
[4] 郑 巍, 谭兴起, 郭良君,等. 鹿茸草的化学成分[J]. 中国天然药物, 2012, 10(2):102-4.[4] Zheng W, Tan X Q, Guo L J, et al. Chemical constituents from Monochasma savatieri[J]. Chin J Nat Med, 2012, 10(2):102-4.
[5] 余 昕, 朱 烨, 欧丽兰,等. 鸭跖草抗炎活性部位筛选及抗炎机制[J]. 中成药, 2015, 37(8):1824-7.[5] Yu X, Zhu Y, Ou L L, et al. Screening of anti-inflammatory active sites and anti-inflammatory mechanism of Commelina communis[J]. Chin Tradit Patent Med, 2015, 37(8):1824-7.
[6] 袁红娥, 周兴栋, 孟令杰,等. 鸭跖草的化学成分研究[J]. 中国中药杂志, 2013, 38(19):3304-8.[6] Yuan H E, Zhou X D, Meng L J,et al. Chemical constituents from Commelina communis[J]. Chin J Chin Mater Med, 2013, 38(19):3304-8.
[7] 纪宝玉, 范崇庆, 裴莉昕,等. 白花蛇舌草的化学成分及药理作用研究进展[J]. 中国实验方剂学, 2014, 20(19):235-40.[7] Ji B Y, Fan C Q, Pei L X, et al. Advance on the chemical and pharmacological effects studies of Hedyotis diffusa[J]. Chin J Exp Tradit Med Form, 2014, 20(19):235-40.
[8] 杨 娴, 刘汝青, 杜德荣,等. 白花蛇舌草总黄酮对黑色素瘤A375和B16细胞增殖和凋亡的影响[J]. 皮肤性病诊疗学杂志, 2011, 18(6):359-62.[8] Yang X, Liu R Q, Du D R, et al. Effects of FOD on the proliferation and apoptosis of melanoma A375 and B16 Cells[J]. J Diagn Ther Dermato-Venereol, 2011, 18(6):359-62.
[9] Wang J H, Shu L H, Yang L L, et al. 2-Hydroxy-3-methylanthraquinone from Hedyotis diffusa WILLD induces apoptosis via alteration of Fas/FasL and activation of caspase-8 in human leukemic THP-1 cells[J]. Arch Med Res, 2011, 42(7):577-83.
[10] 刘 征, 吴洪涛, 倪亚光,等. Wnt5a与炎症信号在炎症性疾病中的交叉互动[J]. 生理学报, 2015, 67(4):437-45.[10] Liu Z, Wu H T, Ni Y G, et al. Crosstalk between Wnt5a and inflammatory signaling in inflammation[J]. Acta Physiol Sin, 2015, 67(4):437-45.
[11] 潘 磊, 金 旺, 潘宏宇,等. 魔芋葡甘露聚糖(KGM)对LPS诱导的THP-1源巨噬细胞产生细胞因子的影响[J]. 中华中医药学刊, 2017, 35(6):1496-501.[11] Pan L, Jin W, Pan H Y, et al. Effect of Konjac Glucomannan ( KGM) on LPS-induced THP-1 derived macrophages produce cytokines[J]. Chin Arch Tradit Chin Med, 2017, 35(6):1496-501.
[12] 袁 琴, 袁 丁, 周志勇,等. 竹节参齐墩果烷皂苷对RAW264.7巨噬细胞SIRT1活性影响及抗炎作用研究[J]. 中国药理学通报, 2016, 32(3):349-54.[12] Yuan Q, Yuan D, Zhou Z Y, et al. Anti-inflammatory effect of Chikusetsu oleanane saponin on RAW264.7 cell through regulating SIRT1 activity[J]. Chin Pharmacol Bull, 2016, 32(3):349-54.
[13] 王晓晨, 吉爱国. NF-κB信号通路与炎症反应[J]. 生理科学进展, 2014, 45(1):68-71.[13] Wang X C, Ji A G. NF-κB signaling pathway and inflammatory response[J]. Prog Physiol Sci, 2014, 45(1):68-71.
[14] 袁 昊, 曾 晖, 肖德明,等. 白藜芦醇通过NF-κB信号通路抑制软骨细胞炎症因子的表达[J]. 中华骨与关节外科杂志, 2016, 9(1):75-9.[14] Yuan H, Zeng H, Xiao D M, et al. Resveratrol inhibits the expression of inflammatory cytokines in cartilage cells through NF-κB signaling pathway[J]. Chin J Bone Joint Surg, 2016, 9(1):75-9.