盐酸小檗碱增强美沙拉嗪在治疗溃疡性结肠炎中的作用机制
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摘要
目的探讨盐酸小檗碱辅助治疗溃疡性结肠炎(UC)的疗效及其作用机制。方法将SD大鼠分为5组(n=8),正常对照组大鼠予自由饮用蒸馏水,其余4组予自由饮用蒸馏水配置的浓度为5.5%的葡聚糖硫酸钠(DSS)溶液,连续7 d,形成UC大鼠模型。第8～14天,美沙拉嗪组、盐酸小檗碱组分别予美沙拉嗪(420 mg/kg)、盐酸小檗碱(100 mg/kg)灌胃,盐酸小檗碱联合美沙拉嗪组予盐酸小檗碱(100 mg/kg)及美沙拉嗪(420 mg/kg)联合灌胃。观察5组大鼠的疾病活动度(DAI评分)以及各组大鼠结肠组织病理学变化,酶联免疫吸附测定试验检测大鼠血清IL-9表达量,逆转录聚合酶链反应方法及免疫组织化学法分别检测大鼠结肠组织中TLR2 mRNA、TLR2蛋白的表达水平。结果 (1)与正常对照组比较,DSS模型组DAI评分、结肠组织病理学评分、IL-9、TLR2 mRNA、TLR2蛋白表达明显升高(P<0.01)。(2)与DSS模型组对比,美沙拉嗪组、盐酸小檗碱组、盐酸小檗碱联合美沙拉嗪组第14天DAI评分、结肠组织病理学评分、血清IL-9、结肠组织TLR2 mRNA、TLR2蛋白表达均下降(P<0.05),且盐酸小檗碱联合美沙拉嗪组下降更明显(P<0.01)。结论盐酸小檗碱可能通过降低IL-9水平以及TLR2 mRNA、TLR2蛋白的表达增强美沙拉嗪治疗UC的作用。
Objective To investigate the effect and mechanism of berberine hydrochloride in the adjuvant treatment of ulcerative colitis(UC). Methods Forty rats were divided into 5 groups(n=8). The normal control group was with given distilled water, while dextran sulfate sodium(DSS) were given in the other 4 groups for the construction of UC model. After modeling, normal control group and DSS model group were given by gavage of distilled water, while mesalazine(420 mg/kg), berberine hydrochloride(100 mg/kg), and berberine hydrochloride(100 mg/kg) combined with mesalazine(420 mg/kg), were given to rats in mesalazine group, berberine hydrochloride, and berberine hydrochloride combined with mesalazine group, respectively. The degree of disease activity(DAI score) and the pathological changes of colon tissue were compared. The serum IL-9 were measured by enzyme linked immunosorbent assay, and the reverse transcription polymerase chain reaction and immunohistochemistry were used to assess the expression levels of TLR2 mRNA and TLR2 protein in rat colon tissues. Results Compared with the normal control group, the DAI score, the colonic histopathology score, IL-9, TLR2 mRNA and TLR2 protein expression in the DSS model group were significantly higher(P<0.01). Compared with the DSS model group, the DAI score, the colonic histopathology score, IL-9, TLR2 mRNA and TLR2 protein expression of mesalazine group,berberine hydrochloride group and berberine hydrochloride combined with mesalazine group were significantly reduced(P<0.05); which were significantly more prominent in berberine hydrochloride combined with mesalazine group(P<0.01). Conclusion Berberine hydrochloride may enhance the effect of methalazine in the treatment of ulcerative colitis by reducing the level of IL-9 and the expression of TLR2 mRNA and TLR2 protein.
Objective To investigate the effect and mechanism of berberine hydrochloride in the adjuvant treatment of ulcerative colitis(UC). Methods Forty rats were divided into 5 groups(n=8). The normal control group was with given distilled water, while dextran sulfate sodium(DSS) were given in the other 4 groups for the construction of UC model. After modeling, normal control group and DSS model group were given by gavage of distilled water, while mesalazine(420 mg/kg), berberine hydrochloride(100 mg/kg), and berberine hydrochloride(100 mg/kg) combined with mesalazine(420 mg/kg), were given to rats in mesalazine group, berberine hydrochloride, and berberine hydrochloride combined with mesalazine group, respectively. The degree of disease activity(DAI score) and the pathological changes of colon tissue were compared. The serum IL-9 were measured by enzyme linked immunosorbent assay, and the reverse transcription polymerase chain reaction and immunohistochemistry were used to assess the expression levels of TLR2 mRNA and TLR2 protein in rat colon tissues. Results Compared with the normal control group, the DAI score, the colonic histopathology score, IL-9, TLR2 mRNA and TLR2 protein expression in the DSS model group were significantly higher(P<0.01). Compared with the DSS model group, the DAI score, the colonic histopathology score, IL-9, TLR2 mRNA and TLR2 protein expression of mesalazine group,berberine hydrochloride group and berberine hydrochloride combined with mesalazine group were significantly reduced(P<0.05); which were significantly more prominent in berberine hydrochloride combined with mesalazine group(P<0.01). Conclusion Berberine hydrochloride may enhance the effect of methalazine in the treatment of ulcerative colitis by reducing the level of IL-9 and the expression of TLR2 mRNA and TLR2 protein.
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)[J].中国中西医结合消化杂志,2018,26(2):105-111,120.
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[3] 左茹,曹雪滨,张文生.黄连素药理作用研究进展[J].环球中医药,2014,7(7):568-572.
[4] Hamonic G,Pasternak JA,Wilson HL.Recognizing conserved non-canonical localization patterns of toll-like receptors in tissues and across species[J].Cell Tiss Res,2018,372(1):1-11.
[5] 潘娜.干酪乳杆菌对RAW264.7细胞和BALB/c小鼠炎症信号通路的调节机制[D].长春:吉林农业大学,2017.
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[7] 刘胜楠,卢雪峰,孙娜,等.Toll样受体2、4及核因子-κB在溃疡性结肠炎黏膜组织中的表达及临床意义[J].山东大学学报:医学版,2012,50(2):60-63.
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[11] Cooper HS,Murthy SN,Shah RS,et al.Clinicopathologic study of dextran sulfate sodium experimental murine colitis[J].Lab Invest,1993,69(2):238-249.
[12] 贾育新,刘喜平,吴建军.溃疡性结肠炎大鼠模型TLR2表达与中医证候相关性研究[J].北京中医药大学学报,2009,32(7):462-464.
[13] 彭艳,刘新春,王晓艳.诱发炎症性肠病的环境危险因素[J].中南大学学报:医学版,2017,42(9):1100-1104.
[14] 张仁霞,傅永锦,梁基智,等.小檗碱治疗大鼠免疫复合性溃疡性结肠炎及下调炎性细胞因子作用[J].临床医学工程,2014,21(11):1406-1409.
[15] 沈雁,钟继红,徐磊,等.盐酸小檗碱对溃疡性结肠炎小鼠结肠组织TNF-α、IL-1β和IL-10表达的影响[J].中国现代应用药学,2017,34(8):1094-1098.
[16] Li YH,Xiao HT,Hu DD,et al.Berberine ameliorates chronic relapsing dextran sulfate sodium-induced colitis in C57BL/6 mice by suppressing Th17 responses[J].Pharmacol Res,2016,110(2):227-239.
[17] 张苏,王志斌,王跃,等.小檗碱在溃疡性结肠炎中作用及机制的研究进展[J].国际消化病杂志,2016,36(3):136-138,193.
[18] Feng AW,Yu C,Mao Q,et al.Berberine hydrochloride attenuates cyclooxygenase-2 expression in rat small intestinal mucosa during acute endotoxemia[J].Fitoterapia,2011,82(7):976-982.
[19] 刘建平,李玲玲,郎晓猛,等.泄浊解毒方对慢性非特异性溃疡性结肠炎大鼠结肠黏膜环氧合酶-2及前列腺素E2的影响[J].河北中医,2015,35(12):1832-1835.
[20] 顾兆伟,赵鹤,曹志伟.过敏性鼻炎小鼠鼻黏膜组织中白介素9及转录因子PU.1表达水平的研究[J].中国全科医学,2016,19(12):1420-1423.
[21] 王冬.Th9细胞和IL-9在TL1A过表达慢性实验性结肠炎中的作用与机制研究[D].石家庄:河北医科大学,2017.
[22] Nalleweg N,Chiriac M T,Podstawa E,et al.IL-9 and its receptor are predominantly involved in the pathogenesis of UC[J].Gut,2015,64(5):743-755.
[23] Siracusa MC,Wojno ED,Artis D.Functional heterogeneity in the basophil cell lineage[J].Adv Immunol,2012,115:141-159.
[24] 屈冬冬,金世禄,刘宝珍,等.IL-9在溃疡性结肠炎患者结肠黏膜中的表达及意义[J].中国当代医药,2016,23(20):37-40.
[25] Moussata D,Goetz M,Gloeckner A,et al.Confocal laser endomicroscopy is a new imaging modality for recognition of intramucosal bacteria in inflammatory bowel disease in vivo[J].Gut,2011,60(1):26-33.
[26] 屈冬冬,金世禄,胡忠波.IL-9/Th9与溃疡性结肠炎的研究进展[J].中国当代医药,2016,23(18):19-21.
[27] Liu Y,Wang Y,Yamakuchi M,et al.Upregulation of toll-like receptor 2 gene expression in macrophage response to peptidoglycan and high concentration of lipopolysaccharide is involved in NF-kappa b activation[J].Infect Immun,2001,69(5):2788-2796.
[28] 荣英蕊,刘建平,郎晓猛,等.泄浊解毒方对溃疡性结肠炎大鼠TLR2及IL-8IL-1β影响的研究[J].四川中医,2012,7(5):33-36.
[29] Wang X,Liu Y,Dong H,et al.Herb-Partitioned Moxibustion Regulates the TLR2/NF-kappaB Signaling Pathway in a Rat Model of Ulcerative Colitis[J].Evid Based Complement Alternat Med,2015,2015:949065.
[2] Yang H,Li Y,Wu W,et al.The incidence of inflammatory bowel disease in Northern China:a prospective population-based study[J].PLoS One,2014,9(7):e101296.
[3] 左茹,曹雪滨,张文生.黄连素药理作用研究进展[J].环球中医药,2014,7(7):568-572.
[4] Hamonic G,Pasternak JA,Wilson HL.Recognizing conserved non-canonical localization patterns of toll-like receptors in tissues and across species[J].Cell Tiss Res,2018,372(1):1-11.
[5] 潘娜.干酪乳杆菌对RAW264.7细胞和BALB/c小鼠炎症信号通路的调节机制[D].长春:吉林农业大学,2017.
[6] Kolar SS,Baidouri H,McDermott AM.Role of Pattern Recognition Receptors in the Modulation of Antimicrobial Peptide Expression in the Corneal Epithelial Innate Response to F.solani[J].Invest Ophthalmol Vis Sci,2017,58(5):2463-2472.
[7] 刘胜楠,卢雪峰,孙娜,等.Toll样受体2、4及核因子-κB在溃疡性结肠炎黏膜组织中的表达及临床意义[J].山东大学学报:医学版,2012,50(2):60-63.
[8] 来利华.模式识别受体触发的巨噬细胞天然免疫应答调控的新机制研究[D].杭州:浙江大学,2013.
[9] 徐叔云,卞如濂,陈修.《药理实验方法学》第二版出版发行[J].中国药理学通报,1992,8(1):19.
[10] Okayasu I,Hatakeyama S,Yamada M,et al.A novel method in the induction of reliable experimental acute and chronic ulcerative colitis in mice[J].Gastroenterology,1990,98(3):694-702.
[11] Cooper HS,Murthy SN,Shah RS,et al.Clinicopathologic study of dextran sulfate sodium experimental murine colitis[J].Lab Invest,1993,69(2):238-249.
[12] 贾育新,刘喜平,吴建军.溃疡性结肠炎大鼠模型TLR2表达与中医证候相关性研究[J].北京中医药大学学报,2009,32(7):462-464.
[13] 彭艳,刘新春,王晓艳.诱发炎症性肠病的环境危险因素[J].中南大学学报:医学版,2017,42(9):1100-1104.
[14] 张仁霞,傅永锦,梁基智,等.小檗碱治疗大鼠免疫复合性溃疡性结肠炎及下调炎性细胞因子作用[J].临床医学工程,2014,21(11):1406-1409.
[15] 沈雁,钟继红,徐磊,等.盐酸小檗碱对溃疡性结肠炎小鼠结肠组织TNF-α、IL-1β和IL-10表达的影响[J].中国现代应用药学,2017,34(8):1094-1098.
[16] Li YH,Xiao HT,Hu DD,et al.Berberine ameliorates chronic relapsing dextran sulfate sodium-induced colitis in C57BL/6 mice by suppressing Th17 responses[J].Pharmacol Res,2016,110(2):227-239.
[17] 张苏,王志斌,王跃,等.小檗碱在溃疡性结肠炎中作用及机制的研究进展[J].国际消化病杂志,2016,36(3):136-138,193.
[18] Feng AW,Yu C,Mao Q,et al.Berberine hydrochloride attenuates cyclooxygenase-2 expression in rat small intestinal mucosa during acute endotoxemia[J].Fitoterapia,2011,82(7):976-982.
[19] 刘建平,李玲玲,郎晓猛,等.泄浊解毒方对慢性非特异性溃疡性结肠炎大鼠结肠黏膜环氧合酶-2及前列腺素E2的影响[J].河北中医,2015,35(12):1832-1835.
[20] 顾兆伟,赵鹤,曹志伟.过敏性鼻炎小鼠鼻黏膜组织中白介素9及转录因子PU.1表达水平的研究[J].中国全科医学,2016,19(12):1420-1423.
[21] 王冬.Th9细胞和IL-9在TL1A过表达慢性实验性结肠炎中的作用与机制研究[D].石家庄:河北医科大学,2017.
[22] Nalleweg N,Chiriac M T,Podstawa E,et al.IL-9 and its receptor are predominantly involved in the pathogenesis of UC[J].Gut,2015,64(5):743-755.
[23] Siracusa MC,Wojno ED,Artis D.Functional heterogeneity in the basophil cell lineage[J].Adv Immunol,2012,115:141-159.
[24] 屈冬冬,金世禄,刘宝珍,等.IL-9在溃疡性结肠炎患者结肠黏膜中的表达及意义[J].中国当代医药,2016,23(20):37-40.
[25] Moussata D,Goetz M,Gloeckner A,et al.Confocal laser endomicroscopy is a new imaging modality for recognition of intramucosal bacteria in inflammatory bowel disease in vivo[J].Gut,2011,60(1):26-33.
[26] 屈冬冬,金世禄,胡忠波.IL-9/Th9与溃疡性结肠炎的研究进展[J].中国当代医药,2016,23(18):19-21.
[27] Liu Y,Wang Y,Yamakuchi M,et al.Upregulation of toll-like receptor 2 gene expression in macrophage response to peptidoglycan and high concentration of lipopolysaccharide is involved in NF-kappa b activation[J].Infect Immun,2001,69(5):2788-2796.
[28] 荣英蕊,刘建平,郎晓猛,等.泄浊解毒方对溃疡性结肠炎大鼠TLR2及IL-8IL-1β影响的研究[J].四川中医,2012,7(5):33-36.
[29] Wang X,Liu Y,Dong H,et al.Herb-Partitioned Moxibustion Regulates the TLR2/NF-kappaB Signaling Pathway in a Rat Model of Ulcerative Colitis[J].Evid Based Complement Alternat Med,2015,2015:949065.