黄龙汤联合美罗培南对CLP模型大鼠PI3K/Akt/NF-κB信号通路调控作用
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摘要
目的:探讨中药黄龙汤对CLP大鼠调控PI3K/Akt/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,分别给予美罗培南、黄龙汤+美罗培南干预3 d。采用ELISA法检测各组大鼠血清中IL-1β、TNF-α、IL-6、IL-8含量;Western-blot法检测回肠组织中PI3K、Akt、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、IL-6、IL-8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PI3K、Akt、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、IL-6、IL-8含量及回肠组织中PI3K、Akt、NF-κB表达水平显著下调,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、IL-6、IL-8的表达,该作用可能是通过调控回肠组织中PI3K/Akt/NF-κB信号通路活化实现的。
Objective:To observe the levels of TNF-α, IL-6 and IL-8 in serum of CPL model rats and the expression levels of PI3 K, Akt and NF-κB in serum of rats after the intervention of Huanglong Decoction combined with meropenem, and investigate the anti inflammatory mechanism of Huanglong Decoction through regulating the PI3 K/Akt/NF-kappa B pathway.Methods:A total of 70 SD rats were treated with cecal ligation and perforation to replicate the CPL rat model. The rats were treated with Huanglong Decoction for 3 days.After 1 hour of the last administration, the whole blood was collected, the serum was separatedand the ileum tissue was cryopreserved.ELISA method was used to detect the contents of IL-1, TNF-α, IL-6 and IL-8 in the serum of each group. The expression levels of PI3 K, Akt and NF-κ B in ileum tissues were detected by Western-blot.Results:Compared with the sham operation group, the serum levels of IL-1β, TNF-α, IL-6 and IL-8 increased significantly in the model control group, and the expression levels of TLR4, Myd88 and NF-κB in the ileum tissue were significantly up-regulated.Compared with the model control group, the contents of IL-1β and TNF-α in the serum of Western medicine group and Huanglong Decoction group were significantly decreased, and the expression levels of PI3 K, Akt and NF-κB in the ileum tissue were significantly down-regulated.Compared with the western medicine group, the levels of serumIL-1β, TNF-α, IL-6, IL-8 and PI3 K, Akt and NF-κB in the intestinal tissue of the Huanglong Decoction combined with western medicine group were significantly down-regulatedand the statistical significance was statistically significant.Conclusion:Huanglong Decoction combined with meropenem has anti-inflammatory effect on CPL model rats and can significantly inhibit the expressions of serum IL-1β, TNF-α, IL-6 and IL-8, which may be realized by regulating the activation of PI3 K/Akt/NF-κB signaling pathway in the ileum.
Objective:To observe the levels of TNF-α, IL-6 and IL-8 in serum of CPL model rats and the expression levels of PI3 K, Akt and NF-κB in serum of rats after the intervention of Huanglong Decoction combined with meropenem, and investigate the anti inflammatory mechanism of Huanglong Decoction through regulating the PI3 K/Akt/NF-kappa B pathway.Methods:A total of 70 SD rats were treated with cecal ligation and perforation to replicate the CPL rat model. The rats were treated with Huanglong Decoction for 3 days.After 1 hour of the last administration, the whole blood was collected, the serum was separatedand the ileum tissue was cryopreserved.ELISA method was used to detect the contents of IL-1, TNF-α, IL-6 and IL-8 in the serum of each group. The expression levels of PI3 K, Akt and NF-κ B in ileum tissues were detected by Western-blot.Results:Compared with the sham operation group, the serum levels of IL-1β, TNF-α, IL-6 and IL-8 increased significantly in the model control group, and the expression levels of TLR4, Myd88 and NF-κB in the ileum tissue were significantly up-regulated.Compared with the model control group, the contents of IL-1β and TNF-α in the serum of Western medicine group and Huanglong Decoction group were significantly decreased, and the expression levels of PI3 K, Akt and NF-κB in the ileum tissue were significantly down-regulated.Compared with the western medicine group, the levels of serumIL-1β, TNF-α, IL-6, IL-8 and PI3 K, Akt and NF-κB in the intestinal tissue of the Huanglong Decoction combined with western medicine group were significantly down-regulatedand the statistical significance was statistically significant.Conclusion:Huanglong Decoction combined with meropenem has anti-inflammatory effect on CPL model rats and can significantly inhibit the expressions of serum IL-1β, TNF-α, IL-6 and IL-8, which may be realized by regulating the activation of PI3 K/Akt/NF-κB signaling pathway in the ileum.
引文
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[18] 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.
[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 Mar 19.[Epub ahead of print].
[5] 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.
[6] 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.
[7] 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.
[8] 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.
[9] 代旭亮.APACHEⅡ评分对ICU脓毒症患者预后的评估及原花青素B1通过PI3K/AKT-NF-KB通路调节THP-1细胞炎症反应的研究[D].大连:大连医科大学,2016.
[10] 向镜芬,杨祥,龚剑锋,等.PI3K/Akt信号通路在脓毒症肾脏损伤诱导的自噬中的调节作用[J].中国病理生理杂志,2014,30(6):1052-1058.
[11] 范婷婷.抑制PI3Kγ表达对脂多糖诱导脓毒症心肌功能障碍小鼠的影响[D].重庆:重庆医科大学,2014.
[12] 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.
[13] Tartey S,Takeuchi O.Pathogen recognition and Toll-like receptor targeted therapeutics in innate immune cells[J].Int Rev Immunol,2017,6:1-17.
[14] 张伟洁,郑宏.IL-6介导免疫炎性反应作用及其与疾病关系的研究进展[J].细胞与分子免疫学杂志,2017,33(5):699-703.
[15] Rostami K,Nejad MR,Asadzadeh H,et al.PTU-199 Proinflammatory Cytokine (Il-8) in Microscopic Enteritis[J].Gut,2013,62(Suppl 1):A130-A130.
[16] Matsushima M,Suzuki T,Masui A,et al.Cranberry extract suppresses interleukin-8 secretion from stomach cells stimulated by Helicobacter pylori,in every clinically separated strain but inhibits growth in part of the strains[J].Journal of Functional Foods,2013,5(2):729-735.
[17] Wehrle J,Seeger TS,Schwemmers S,et al.Transcription factor nuclear factor erythroid-2 mediates expression of the cytokine interleukin 8,a known predictor of inferior outcome in patients with myeloproliferative neoplasms[J].Haematologica,2013,98(7):1073-1080.
[18] 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.