中药黄龙汤对CPL大鼠回肠组织中TLR4/NF-κB信号通路调控的实验研究
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摘要
目的:通过检测中药黄龙汤干预后CPL模型大鼠中血清中IL-1β、TNF-α含量,回肠组织TLR4、Myd88、NF-κB表达水平,探讨中药黄龙汤通过调控TLR4/NF-κB通路途径的抗炎作用机制。方法:SD大鼠70只,采用盲肠结扎穿孔术复制CPL大鼠模型,并经口给予中药黄龙汤干预,连续干预3 d。末次给药后1 h,采集全血,分离血清,同时取回肠组织冻存。采用ELISA法检测各组大鼠血清中IL-1β、TNF-α含量;Western-blot法检测回肠组织中TLR4、Myd88、NF-κB表达水平。结果:与假手术组比较,模型对照组大鼠血清IL-1β、TNF-α含量显著升高,回肠组织中TLR4、Myd88、NF-κB表达水平显著上调;与模型对照组比较,西药组和黄龙汤联合西药组大鼠血清IL-1β、TNF-α含量显著降低,回肠组织中TLR4、Myd88、NF-κB表达水平显著下调;与西药组比较,黄龙汤联合西药组大鼠血清IL-1β、TNF-α含量及回肠组织中TLR4、Myd88、NF-κB表达水平显著下调,差异有统计学意义。结论:黄龙汤联合西药美罗培南对CPL模型大鼠具有抗炎作用,能够显著抑制血清IL-1β、TNF-α的表达,该作用可能是通过调控回肠组织中TLR4/NF-κB信号通路活化实现的。
Objective: By detecting the IL-1 beta and TNF-alpha levels in serum and the expression levels of TLR4, Myd88 and NF-κB in the ileum tissues of CPL rats after the intervention of Huanglong Decoction, to explore anti-inflammatory mechanism of Huanglong Decoction through regulating TLR4/NF-κB pathway.Methods: Seventy SD rats were used to replicate CPL rat model with cecal ligation perforation, and the traditional Chinese medicine Huanglong Decoction was given orally. The intervention lasted for 3 days. One hour after the last administration, the whole blood was collected, the serum was isolated, and the ileum was collected and frozen. The serum levels of IL-1 and TNF-alpha in rats were detected by ELISA. The expression levels of TLR4, Myd88 and NF-κB were detected by Westernblot method. Results: Compared with the sham operation group, serumIL-1β and TNF-αof rats in the control group were significantly increased, and TLR4, Myd88 and NF-κB expression levels were significantly increased in ileal tissues. Compared with the control group, the contents of serum IL-1β and TNF-αwere significantly reduced in the western medicine group and the Huanglong Decoction, and TLR4, Myd88 and NF-κB were significantly reduced in the ileum. Compared with the western medicine group, the levels of serum IL-1β, TNF-α and the TLR4, Myd88 and NF-κB expression levels of the rats in the combined western medicine group were significantly reduced, which was statistically significant. Conclusion: Huanglong Decoction combined western medicine meropenem on CPL model rats have anti-inflammatory effect and can significantly inhibit the serum IL-1β and the expression of TNF-α. It could be a role by regulating the ileum tissue TLR4/NF-κB signaling pathway activation.
Objective: By detecting the IL-1 beta and TNF-alpha levels in serum and the expression levels of TLR4, Myd88 and NF-κB in the ileum tissues of CPL rats after the intervention of Huanglong Decoction, to explore anti-inflammatory mechanism of Huanglong Decoction through regulating TLR4/NF-κB pathway.Methods: Seventy SD rats were used to replicate CPL rat model with cecal ligation perforation, and the traditional Chinese medicine Huanglong Decoction was given orally. The intervention lasted for 3 days. One hour after the last administration, the whole blood was collected, the serum was isolated, and the ileum was collected and frozen. The serum levels of IL-1 and TNF-alpha in rats were detected by ELISA. The expression levels of TLR4, Myd88 and NF-κB were detected by Westernblot method. Results: Compared with the sham operation group, serumIL-1β and TNF-αof rats in the control group were significantly increased, and TLR4, Myd88 and NF-κB expression levels were significantly increased in ileal tissues. Compared with the control group, the contents of serum IL-1β and TNF-αwere significantly reduced in the western medicine group and the Huanglong Decoction, and TLR4, Myd88 and NF-κB were significantly reduced in the ileum. Compared with the western medicine group, the levels of serum IL-1β, TNF-α and the TLR4, Myd88 and NF-κB expression levels of the rats in the combined western medicine group were significantly reduced, which was statistically significant. Conclusion: Huanglong Decoction combined western medicine meropenem on CPL model rats have anti-inflammatory effect and can significantly inhibit the serum IL-1β and the expression of TNF-α. It could be a role by regulating the ileum tissue TLR4/NF-κB signaling pathway activation.
引文
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[11] 郑福奎,唐农,吴林,等.清瘟败毒饮对脓毒症相关性脑病大鼠脑组织与TLR4介导的炎性因子的影响[J].广西中医药,2017,40(2):70-73.
[12] Sun J,Shi S,Wang Q,et al.Continuous hemodiafiltration therapy reduces damage of multi-organs by ameliorating of HMGB1/TLR4/NFκB in a dog sepsis model[J].Int J Clin Exp Pathol,2015,8(2):1555-1564.
[13] 凌羽,张子琪,兰海楠,等.虾青素对脂多糖通过TLR4/MyD88/NF-κB信号通路诱导的IPEC-J2细胞炎症的影响[J].华南农业大学学报,2018(5):1-6.
[14] 周丽雅,柴晶美,杨阿龙,等.上中下通用痛风汤对大鼠急性痛风关节炎模型TLR/MYD88信号通路影响的研究[J].中国中医基础医学杂志,2018,24(5):604-606.
[15] 朱怡,李毅,张楠,等.电针预处理对心肌缺血再灌注损伤大鼠“内关”穴区Toll样受体4、髓样分化因子88及核转录因子-κB基因表达的影响[J].针刺研究,2018,43(5):302-306,329.
[16] 庞高举,孙丽妲,姚楠,等.TLR2、TLR4及MyD88高表达与Cm呼吸道感染肺组织炎症相关[J].中国免疫学杂志,2018,34(5):737-740.
[17] Liu M,Li C,Zhao GQ,et al.Boxb mediate BALB/c mice corneal inflammation through a TLR4/MyD88-dependent signaling pathway in Aspergillus fumigateskeratiti[J].International Journal of Ophthalmology,2018,11(4):548-552.
[18] YANG YL,CHENG X,LI WH,et al.KAE Alleviates LPS-induced Neuroinflammation in the Striatum of Mice via Down-regulating TLR4/MyD88 Signaling Pathway[J].神经药理学报,2018,8(2):30.
[19] 王创畅,吴伟,魏伟超,等.清热、活血中药调控TLR4/MyD88/NF-κB信号干预动脉粥样硬化大鼠模型的实验研究[J].中华中医药学刊,2018,36(3):576-580.
[20] 李丹丹,裴丽霞,耿昊,等.基于TLR2/MyD88/NF-κB信号通路探讨针灸治疗肠易激综合征的机制[J].辽宁中医杂志,2018,45(2):366-369.
[2] 高君伟. 创伤后脓毒症遗传易感性的流行病学和临床关联性研究[D].重庆:第三军医大学,2016.
[3] Rhodes A,Phillips G,Beale R,et al.The surviving sepsis campaign bundles and outcome:results from the International Multicentre Prevalence Study on Sepsis(the IMPreS S study)[J].Intensive Care Med,2015,41(9):1620-1628.
[4] Lakshmikanth CL,Jacob SP,Chaithra VH,et al.Sepsis:in search of cure[J].Inflamm Res,2016(3):310-312.
[5] 付瑜,黄煜,姜树民.黄龙汤对脓毒症大鼠肠道黏膜免疫屏障保护作用[J].辽宁中医药大学学报,2017,19(7):39-42.
[6] Bellani G, Laffey JG, Pham T, et al.Epidemiology, Patterns of Care, and Mortality for Patients With Acute Respiratory Distress Syndrome in Intensive Care Units in 50 Countries[J].JAMA, 2016, 315(8):788-800.
[7] Sun J, Shi S, Wang Q, et al.Continuous hemodiafiltration therapy reduces damage of multi-organs by ameliorating of HMGB1/TLR4/NFκB in a dog sepsis model[J].Int J Clin Exp Pathol, 2015, 8(2):1555-1564.
[8] 王华柱,陈建丽,徐艳霞,等.脓毒症及脓毒性休克大鼠脑细胞TLR4和TNF-α的表达及细胞凋亡研究[J].贵州医药,2018,42(1):3-5.
[9] 朱滨,吴叶顺.脓毒症导致ALI/ARDS患者外周血TLR4及血清TNF-α、SP-A的变化及相关性分析[J].中国医药指南,2017,15(31):1-2.
[10] 张建峰,刘涛,曹波,等.右美托咪定通过调控HMGB1/TLR4/NF-κB信号通路抗脓毒症大鼠肺损伤的实验研究[J].临床肺科杂志,2017,22(10):1760-1763.
[11] 郑福奎,唐农,吴林,等.清瘟败毒饮对脓毒症相关性脑病大鼠脑组织与TLR4介导的炎性因子的影响[J].广西中医药,2017,40(2):70-73.
[12] Sun J,Shi S,Wang Q,et al.Continuous hemodiafiltration therapy reduces damage of multi-organs by ameliorating of HMGB1/TLR4/NFκB in a dog sepsis model[J].Int J Clin Exp Pathol,2015,8(2):1555-1564.
[13] 凌羽,张子琪,兰海楠,等.虾青素对脂多糖通过TLR4/MyD88/NF-κB信号通路诱导的IPEC-J2细胞炎症的影响[J].华南农业大学学报,2018(5):1-6.
[14] 周丽雅,柴晶美,杨阿龙,等.上中下通用痛风汤对大鼠急性痛风关节炎模型TLR/MYD88信号通路影响的研究[J].中国中医基础医学杂志,2018,24(5):604-606.
[15] 朱怡,李毅,张楠,等.电针预处理对心肌缺血再灌注损伤大鼠“内关”穴区Toll样受体4、髓样分化因子88及核转录因子-κB基因表达的影响[J].针刺研究,2018,43(5):302-306,329.
[16] 庞高举,孙丽妲,姚楠,等.TLR2、TLR4及MyD88高表达与Cm呼吸道感染肺组织炎症相关[J].中国免疫学杂志,2018,34(5):737-740.
[17] Liu M,Li C,Zhao GQ,et al.Boxb mediate BALB/c mice corneal inflammation through a TLR4/MyD88-dependent signaling pathway in Aspergillus fumigateskeratiti[J].International Journal of Ophthalmology,2018,11(4):548-552.
[18] YANG YL,CHENG X,LI WH,et al.KAE Alleviates LPS-induced Neuroinflammation in the Striatum of Mice via Down-regulating TLR4/MyD88 Signaling Pathway[J].神经药理学报,2018,8(2):30.
[19] 王创畅,吴伟,魏伟超,等.清热、活血中药调控TLR4/MyD88/NF-κB信号干预动脉粥样硬化大鼠模型的实验研究[J].中华中医药学刊,2018,36(3):576-580.
[20] 李丹丹,裴丽霞,耿昊,等.基于TLR2/MyD88/NF-κB信号通路探讨针灸治疗肠易激综合征的机制[J].辽宁中医杂志,2018,45(2):366-369.