清肠温中方促进DSS诱导溃疡性结肠炎大鼠结肠claudin-l、claudin-4表达修复肠黏膜屏障的机制研究
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摘要
目的:研究清肠温中方对葡聚糖硫酸钠(DSS)诱导溃疡性结肠炎(UC)大鼠肠黏膜屏障的影响。方法:采用4.5%DSS溶液制备大鼠UC模型,给予清肠温中方及美沙拉嗪灌胃,Western blot、Real time-PCR检测结肠claudin-1、claudin-4的表达;应用免疫组织化学法检测NF-κB p65蛋白的表达及分布。结果:与模型组相比,清肠温中方和美沙拉嗪组大鼠结肠claudin-1、claudin-4的基因和蛋白表达水平均显著升高(P <0.05),NF-κB p65平均光密度显著降低(P <0.05)。结论:清肠温中方可能通过下调NF-κB p65的表达,促进claudin-l、claudin-4的表达,修复肠粘膜损伤,最终达到治疗UC的目的。
Objective: The purpose of this paper is to study the effect of Qingchang Wenzhong Decoction on intestinal mucosal barrier in the rats with DSS-induced ulcerative colitis(UC) Methods: the UC model was made using 4.5%(w/v)DSS liquor. Meanwhile, the gavage of Qingchang Wenzhong decoction and mesalazine was given. Claudin-1 and claudin-4 expression were detected by Western blot and Real time-PCR. The expression and distribution of NF-κB p65 protein were detected by immunohistochemistry.Results: compared with the model group, the genes and protein levels of claudinl and claudin-4 of colitis rats with Qingchang Wenzhong decoction and mesalazine group significantly increased(P <0.05), and the average optical density of NF-κB p65 significantly decreased(P < 0.05)Conclusion: Qingchang Wenzhong decoction could promote the expression claudin-l and claudin-4 by reducing the expression of NF-κB p65, repairing intestinal mucosal damage and finally treating UC.
Objective: The purpose of this paper is to study the effect of Qingchang Wenzhong Decoction on intestinal mucosal barrier in the rats with DSS-induced ulcerative colitis(UC) Methods: the UC model was made using 4.5%(w/v)DSS liquor. Meanwhile, the gavage of Qingchang Wenzhong decoction and mesalazine was given. Claudin-1 and claudin-4 expression were detected by Western blot and Real time-PCR. The expression and distribution of NF-κB p65 protein were detected by immunohistochemistry.Results: compared with the model group, the genes and protein levels of claudinl and claudin-4 of colitis rats with Qingchang Wenzhong decoction and mesalazine group significantly increased(P <0.05), and the average optical density of NF-κB p65 significantly decreased(P < 0.05)Conclusion: Qingchang Wenzhong decoction could promote the expression claudin-l and claudin-4 by reducing the expression of NF-κB p65, repairing intestinal mucosal damage and finally treating UC.
引文
1李军祥,陈誩.溃疡性结肠炎中西医结合诊疗共识意见(2017年).中国中西医结合消化杂志,2018(2):105-111+120.
2 Sclano G,Asthma,nasal polyposis and ulcerative colitis:a new perspective.Clin Exp Allergy.2002,32(8):1144-1149.
3 Jess T,Rungoe C,Peyrin-Biroulet L.Risk of colorectal cancer in patients with ulcerative colitis:a meta-analysis of population-based cohort studies.Clin Gastroenterol Hepatol.2012,10(6):639-645.
4 Ananthakrishnan AN.Epidemiology and risk factors for IBD.Nat Rev Gastroenterol Hepatol.2015,12(4):205-217
5王福彦.炎症性肠病的发病率、患病率与环境因素的影响.中国社会医学杂志,2007,24(2):138-139.
6 Prideaux L,Kamm M A,De Cruz P P,et al.Inflammatory bowel disease in Asia:a systematic review.J Gastroenterol Hepatol.2012,27(8):1266-1280.
7 Landy J,Ronde E,English N,et al.Tight junctions in inflammatory bowel diseases and inflammatory bowel disease associated colorectal cancer.World J Gastroenterol.2016,22(11):3117-26.
8毛堂友,程佳伟,魏仕兵,等.清肠温中方治疗溃疡性结肠炎84例环球中医药,2016,9(4):479-481.
9王志斌,陈晨,郭一,等.清肠温中方治疗轻中度溃疡性结肠炎的临床研究.中国中西医结合杂志,2018,38(1):15-19.
10 Mao T Y,Rui Shi,Weihan Zhao,et al.Qingchang Wenzhong Decoction Ameliorates Dextran Sulphate Sodium-Induced Ulcerative Colitis in Rats by Downregulating the IP10/CXCR3 Axis-Mediated Inflammatory Response,Evid Based Complement Alternat Med,2016,2016(4312538).
11 Deng J,Zeng L,Lai X,et al.Metformin protects against intestinal barrier dysfunction via AMPKα1-dependent inhibition of JNK signalling activation.J Cell Mol Med.2018,22(1):546-557.
12 S Bron P A,Kleerebezem M,Brummer R J,et al.Can probiotics modulate human disease by impacting intestinal barrier function?.Br J Nutr2017,117(1):93-107.
13 Salim S Y,S?derholm J D.Importance of disrupted intestinal barrier in inflammatory bowel diseases.Inflamm Bowel Dis,2011,17:362-381.
14 Michielan A,D’IncàR.Intestinal permeability in inflammatory bowel disease:pathogenesis,clinical evaluation,and therapy of leaky gut.Mediators Inflamm.2015;2015:628157
15 Das P,Goswami P,Das T K,et al.Comparative tight junction protein expressions in colonic Crohn.s disease,ulcerative colitis,and tuberculosis:a new perspective.Virchows Arch.2012;460(3):261-270.
16 Edelblum K L,Turner J R.The tight junction in inflammatory disease:communication breakdown.Curr Opin Pharmacol,2009,9(6):715-720.
17 Groschwitz K R,Hogan S P.Intestinal barrier function:molecular regulation and disease pathogenesis.J Allergy Clin Immunol,2009,124:3-20.
18 Landy J,Ronde E,English N,et al.Tight junctions in inflammatory bowel diseases and inflammatory bowel disease associated colorectal cancer.World J Gastroenterol.2016,22(11):3117-26.
19马军宏,于向阳.紧密连接蛋白与肠黏膜屏障损伤研究进.中国中西医结合外科杂志,2015,21(1):104.
20向若兰,苏运超,裴晓庆,吴立玲.紧密连接蛋白claudin-4的研究进展.生理科学进展,2012,43(04):310-314.
21 Liu Y,Liu X,Hua W,et al.Berberine inhibits macrophage M1 polarization via AKT1/SOCS1/NF-κB signaling pathway to protect against DSS-induced colitis.Int Immunopharmacol.2018,57:121-131.
22 Gao Z,Yu C,Liang H,Wang X,et al.Andrographolide derivative CX-10 ameliorates dextran sulphate sodium-induced ulcerative colitis in mice:Involvement of NF-κB and MAPK signalling pathways.Int Immunopharmacol.2018,57:82-90.
23 Akanda M R,Nam H H,Tian W,et al.Regulation of JAK2/STAT3 and NF-κB signal transduction pathways;Veronica polita alleviates dextran sulfate sodium-induced murine colitis.Biomed Pharmacother.2018,12;100:296-303.
24 Kaneko Y,Kohno T,Kakuki T,et al.The role of transcriptional factor p63 in regulation of epithelial barrier and ciliogenesis of human nasal epithelial cells. Sci Rep. 2017, 7(1):10935.
2 Sclano G,Asthma,nasal polyposis and ulcerative colitis:a new perspective.Clin Exp Allergy.2002,32(8):1144-1149.
3 Jess T,Rungoe C,Peyrin-Biroulet L.Risk of colorectal cancer in patients with ulcerative colitis:a meta-analysis of population-based cohort studies.Clin Gastroenterol Hepatol.2012,10(6):639-645.
4 Ananthakrishnan AN.Epidemiology and risk factors for IBD.Nat Rev Gastroenterol Hepatol.2015,12(4):205-217
5王福彦.炎症性肠病的发病率、患病率与环境因素的影响.中国社会医学杂志,2007,24(2):138-139.
6 Prideaux L,Kamm M A,De Cruz P P,et al.Inflammatory bowel disease in Asia:a systematic review.J Gastroenterol Hepatol.2012,27(8):1266-1280.
7 Landy J,Ronde E,English N,et al.Tight junctions in inflammatory bowel diseases and inflammatory bowel disease associated colorectal cancer.World J Gastroenterol.2016,22(11):3117-26.
8毛堂友,程佳伟,魏仕兵,等.清肠温中方治疗溃疡性结肠炎84例环球中医药,2016,9(4):479-481.
9王志斌,陈晨,郭一,等.清肠温中方治疗轻中度溃疡性结肠炎的临床研究.中国中西医结合杂志,2018,38(1):15-19.
10 Mao T Y,Rui Shi,Weihan Zhao,et al.Qingchang Wenzhong Decoction Ameliorates Dextran Sulphate Sodium-Induced Ulcerative Colitis in Rats by Downregulating the IP10/CXCR3 Axis-Mediated Inflammatory Response,Evid Based Complement Alternat Med,2016,2016(4312538).
11 Deng J,Zeng L,Lai X,et al.Metformin protects against intestinal barrier dysfunction via AMPKα1-dependent inhibition of JNK signalling activation.J Cell Mol Med.2018,22(1):546-557.
12 S Bron P A,Kleerebezem M,Brummer R J,et al.Can probiotics modulate human disease by impacting intestinal barrier function?.Br J Nutr2017,117(1):93-107.
13 Salim S Y,S?derholm J D.Importance of disrupted intestinal barrier in inflammatory bowel diseases.Inflamm Bowel Dis,2011,17:362-381.
14 Michielan A,D’IncàR.Intestinal permeability in inflammatory bowel disease:pathogenesis,clinical evaluation,and therapy of leaky gut.Mediators Inflamm.2015;2015:628157
15 Das P,Goswami P,Das T K,et al.Comparative tight junction protein expressions in colonic Crohn.s disease,ulcerative colitis,and tuberculosis:a new perspective.Virchows Arch.2012;460(3):261-270.
16 Edelblum K L,Turner J R.The tight junction in inflammatory disease:communication breakdown.Curr Opin Pharmacol,2009,9(6):715-720.
17 Groschwitz K R,Hogan S P.Intestinal barrier function:molecular regulation and disease pathogenesis.J Allergy Clin Immunol,2009,124:3-20.
18 Landy J,Ronde E,English N,et al.Tight junctions in inflammatory bowel diseases and inflammatory bowel disease associated colorectal cancer.World J Gastroenterol.2016,22(11):3117-26.
19马军宏,于向阳.紧密连接蛋白与肠黏膜屏障损伤研究进.中国中西医结合外科杂志,2015,21(1):104.
20向若兰,苏运超,裴晓庆,吴立玲.紧密连接蛋白claudin-4的研究进展.生理科学进展,2012,43(04):310-314.
21 Liu Y,Liu X,Hua W,et al.Berberine inhibits macrophage M1 polarization via AKT1/SOCS1/NF-κB signaling pathway to protect against DSS-induced colitis.Int Immunopharmacol.2018,57:121-131.
22 Gao Z,Yu C,Liang H,Wang X,et al.Andrographolide derivative CX-10 ameliorates dextran sulphate sodium-induced ulcerative colitis in mice:Involvement of NF-κB and MAPK signalling pathways.Int Immunopharmacol.2018,57:82-90.
23 Akanda M R,Nam H H,Tian W,et al.Regulation of JAK2/STAT3 and NF-κB signal transduction pathways;Veronica polita alleviates dextran sulfate sodium-induced murine colitis.Biomed Pharmacother.2018,12;100:296-303.
24 Kaneko Y,Kohno T,Kakuki T,et al.The role of transcriptional factor p63 in regulation of epithelial barrier and ciliogenesis of human nasal epithelial cells. Sci Rep. 2017, 7(1):10935.