参苓白术散通过调节肠上皮细胞自噬治疗葡聚糖硫酸钠所致小鼠炎症性肠病
|
摘要
目的:探究参苓白术散对5%葡聚糖硫酸钠(DSS)所致BALB/c小鼠炎症性肠病(IBD)的改善作用与肠上皮细胞自噬的关系。方法:84只BALB/c小鼠,除正常组之外,5%DSS自由饮用7 d诱导急性炎症性肠病,治疗组分别给予参苓白术散高、中、低剂量组(12,6,3 g·kg-1·d-1),美沙拉嗪组(2 g·kg-1·d-1),自噬诱导剂雷帕霉素组(4 mg·kg-1·d-1)灌胃。观察小鼠体质量、粪便性状、隐血便血,计算疾病活动度(DAI)积分;酶联免疫吸附测定(ELISA)检测小鼠血清中炎症因子白细胞介素-8(IL-8),IL-10含量;苏木素-伊红(HE)染色观察结肠组织病理学改变;免疫组化检测肠组织中肿瘤坏死因子-α(TNF-α)和IL-1β的表达;共聚焦显微镜观察肠上皮细胞自噬体形成情况;蛋白免疫印迹法(Western blot)检测自噬相关通路蛋白的水平。结果:与模型组比较,参苓白术散组小鼠隐血,便血,体质量下降,DAI积分,病理改变等均改善;与正常组比较,模型组小鼠血清中IL-10含量显著降低,IL-8含量显著升高(P <0. 01),小鼠肠组织中IL-1β和TNF-α的表达明显升高(P <0. 01),肠上皮细胞自噬体减少,微管轻链蛋白3(LC3)-Ⅱ/LC3-Ⅰ比值含量显著降低(P <0. 01);与模型组比较,参苓白术散可以升高小鼠血清IL-10含量,降低IL-8含量明显(P <0. 05,P <0. 01),明显降低IL-1β和TNF-α的表达(P <0. 05),参芩白术散高剂量组、雷帕霉素及美沙拉嗪组肠上皮细胞自噬体形成显著增多,显著升高LC3Ⅱ/LC3Ⅰ含量。结论:参苓白术散抗DSS诱导的IBD的作用与抑制炎症及调节肠上皮细胞自噬有关。
Objective: To investigate the mechanism of Shenling Baizhu San(SLBZS) in treating dextra sulfate sodium(DSS)-induced inflammatory bowel disease(IBD) mice and its relationship with autophagy.Method: SPF BALB/c mice were randomly divided into control group,model group,mesalazine group,low,medium and high-dose SLBZS groups,and autophagy inducer rapamycin group. The IBD mice were fed with 5%DSS in their drinking water for 7 days,and the control mice received only water. SLBZS groups were given SLBZS at doses of 3,6,12 g·kg-1·d-1,positive group was given mesalazine sustained release granules at the dose of2 g·kg-1·d-1,rapamycin group was given rapamycin at the dose of 4 mg·kg-1·d-1,and control mice was given the same volume of normal saline by gavage. The mice weight,stool occult blood in stool,score of disease activity(DAI),pathological examination of intestinal mucosal lesions integral were observed after 7 days. interleukin(IL)-8 and IL-10 in serum were detected by enzyme-linked immuno sorbent assay(ELISA),vascular tissue samples were prepared for the detection of tumor neorosis factor-(TNF-α) and IL-1β,and transmission electron microscope and Western blot were used to detect the formation of autophagosomes and the level of autophagy.Result: The body mass decrease,the colon length,disease activity scoring,and histological scoring of SLBZS group were better than those of DSS group. Compared with control group,the level of IL-10 decreased,while the level of IL-8 increased obviously(P < 0. 01),IL-1 and TNF-α expressions significantly up-regulated(P < 0. 01),the formation of autophagosome decreased,and LC3-Ⅱ/Ⅰ level down-regulated. Compared with IBD group,SLBZS group showed increase in formation of autophagosome and LC3-Ⅱ/Ⅰ and IL-10,decrease in IL-8(P <0. 05,P < 0. 01),and down-regulation in IL-1 and TNF-α expressions(P < 0. 01). These results were observed in mesalazine group and rapamycin group. Conclusion: Shenling Baizhu San can significantly inhibit the IBD by regulating autophagy and suppressing inflammation.
Objective: To investigate the mechanism of Shenling Baizhu San(SLBZS) in treating dextra sulfate sodium(DSS)-induced inflammatory bowel disease(IBD) mice and its relationship with autophagy.Method: SPF BALB/c mice were randomly divided into control group,model group,mesalazine group,low,medium and high-dose SLBZS groups,and autophagy inducer rapamycin group. The IBD mice were fed with 5%DSS in their drinking water for 7 days,and the control mice received only water. SLBZS groups were given SLBZS at doses of 3,6,12 g·kg-1·d-1,positive group was given mesalazine sustained release granules at the dose of2 g·kg-1·d-1,rapamycin group was given rapamycin at the dose of 4 mg·kg-1·d-1,and control mice was given the same volume of normal saline by gavage. The mice weight,stool occult blood in stool,score of disease activity(DAI),pathological examination of intestinal mucosal lesions integral were observed after 7 days. interleukin(IL)-8 and IL-10 in serum were detected by enzyme-linked immuno sorbent assay(ELISA),vascular tissue samples were prepared for the detection of tumor neorosis factor-(TNF-α) and IL-1β,and transmission electron microscope and Western blot were used to detect the formation of autophagosomes and the level of autophagy.Result: The body mass decrease,the colon length,disease activity scoring,and histological scoring of SLBZS group were better than those of DSS group. Compared with control group,the level of IL-10 decreased,while the level of IL-8 increased obviously(P < 0. 01),IL-1 and TNF-α expressions significantly up-regulated(P < 0. 01),the formation of autophagosome decreased,and LC3-Ⅱ/Ⅰ level down-regulated. Compared with IBD group,SLBZS group showed increase in formation of autophagosome and LC3-Ⅱ/Ⅰ and IL-10,decrease in IL-8(P <0. 05,P < 0. 01),and down-regulation in IL-1 and TNF-α expressions(P < 0. 01). These results were observed in mesalazine group and rapamycin group. Conclusion: Shenling Baizhu San can significantly inhibit the IBD by regulating autophagy and suppressing inflammation.
引文
[1] Singh S,Chowdhry M,Umar S,et al. Variations in the medical treatment of inflammatory bowel disease among gastroenterologists[J]. Gastroenterol Rep,2018,6(1):61-64.
[2] LIU Y H, DING Y, GAO C C,et al. Functional macrophages and gastrointestinal disorders[J]. World J Gastroenterol,2018,24(11):1181-1195.
[3] Martini E,Krug S M,Siegmund B,et al. Mend your fences:the epithelial barrier and its relationship with mucosal immunity in inflammatory bowel disease[J].Cell Mol Gas Hep,2017,4(1):33-46.
[4]张琮,王嘉正,沈玉洁,等.自噬诱导剂pp242对肿瘤坏死因子-α所致肠屏障损伤的影响[J].胃肠病学,2017,22(6):337-340.
[5]陆忠凯,陈志荣.炎症性肠病患者自噬相关IRGM基因的表达变化及临床意义[J].胃肠病学和肝病学杂志,2013,22(10):1010-1012.
[6]徐昌君,王鹏飞,刘杨,等.黄芪甲苷对特发性肺纤维化自噬活性作用及PI3K/Akt/m TOR信号调控的影响[J].中国实验方剂学杂志,2017,23(18):75-82.
[7]焦延娜,韩淑燕.抗癌中药单体对肿瘤细胞自噬的调控[J].中国实验方剂学杂志,2017,23(7):206-214.
[8]刘玉晖,胡婕,易文凤,等.参苓白术散治疗炎症性肠病与肠上皮细胞紧密连接的关系探讨[J].中国实验方剂学杂志,2015,21(3):130-133.
[9]刘玉晖,刘志勇,廖旺娣,等.参苓白术散抗脂多糖致肠隐窝上皮细胞损伤的作用及机制[J].中药新药与临床药理,2016,27(1):1-5.
[10] YOU Y,FU J J,MENG J, et al. Effect of Nacetylcysteine on the murine model of colitis induced by dextran sodium sulfate through up-regulating PON1activity[J]. Dig Dis Sci,2009,54(8):1643-1650.
[11] YEN Y H,PU C M,LIU C W,et al. Curcumin accelerates cutaneous wound healing via multiple biological actions:the involvement of TNF-α,MMP-9,α-SMA,and collagen[J]. Int Wound J,2018,5(4):605-617.
[12] Majumder K,Mine Y,WU J. The potential of food protein-derived anti-inflammatory peptides against various chronic inflammatory diseases[J]. J Sci Food Agric,2016,96(7):2303-2311.
[13] Manresa M C,Taylor C T. Hypoxia inducible factor(HIF)hydroxylases as regulators of intestinal epithelial barrier function[J]. Cell Mol Gas Hep,2017,3(3):303-315.
[14]郑晴,包怡敏.中药对心肌缺血再灌注损伤过程中自噬的调控[J].中国中药杂志,2017,42(15):2925-2929.
[15]涂玥,孙伟,陈涤平,等.自噬调控肾脏衰老的分子机制及中药的干预作用[J].中国中药杂志,2016,41(21):3914-3918.
[16] Serbati N,Senhaji N,Diakite B,et al. IL23R and ATG16L1 variants in moroccan patients with inflammatory bowel disease[J]. BMC Res Notes,2014,26(7):570.
[17] Baskaran K, Pugazhendhi S, Ramakrishna B S.Association of IRGM gene mutations with inflammatory bowel disease in the Indian population[J]. PLo S One,2014,9(9):e106863.
[18] Paul S,Kashyap A K,JIA W,et al. Selective autophagy of the adaptor protein Bcl10 modulates T cell receptor activation of NF-κB[J]. Immunity,2012,36(6):947-958.
[19] Castillo F,Dekonenko A,Arko-Mensah J,et al.Autophagy protects against active tuberculosis by suppressing bacterial burden and inflammation[J]. Proc Natl Acad Sci USA,2012,109(46):3168-3176.
[2] LIU Y H, DING Y, GAO C C,et al. Functional macrophages and gastrointestinal disorders[J]. World J Gastroenterol,2018,24(11):1181-1195.
[3] Martini E,Krug S M,Siegmund B,et al. Mend your fences:the epithelial barrier and its relationship with mucosal immunity in inflammatory bowel disease[J].Cell Mol Gas Hep,2017,4(1):33-46.
[4]张琮,王嘉正,沈玉洁,等.自噬诱导剂pp242对肿瘤坏死因子-α所致肠屏障损伤的影响[J].胃肠病学,2017,22(6):337-340.
[5]陆忠凯,陈志荣.炎症性肠病患者自噬相关IRGM基因的表达变化及临床意义[J].胃肠病学和肝病学杂志,2013,22(10):1010-1012.
[6]徐昌君,王鹏飞,刘杨,等.黄芪甲苷对特发性肺纤维化自噬活性作用及PI3K/Akt/m TOR信号调控的影响[J].中国实验方剂学杂志,2017,23(18):75-82.
[7]焦延娜,韩淑燕.抗癌中药单体对肿瘤细胞自噬的调控[J].中国实验方剂学杂志,2017,23(7):206-214.
[8]刘玉晖,胡婕,易文凤,等.参苓白术散治疗炎症性肠病与肠上皮细胞紧密连接的关系探讨[J].中国实验方剂学杂志,2015,21(3):130-133.
[9]刘玉晖,刘志勇,廖旺娣,等.参苓白术散抗脂多糖致肠隐窝上皮细胞损伤的作用及机制[J].中药新药与临床药理,2016,27(1):1-5.
[10] YOU Y,FU J J,MENG J, et al. Effect of Nacetylcysteine on the murine model of colitis induced by dextran sodium sulfate through up-regulating PON1activity[J]. Dig Dis Sci,2009,54(8):1643-1650.
[11] YEN Y H,PU C M,LIU C W,et al. Curcumin accelerates cutaneous wound healing via multiple biological actions:the involvement of TNF-α,MMP-9,α-SMA,and collagen[J]. Int Wound J,2018,5(4):605-617.
[12] Majumder K,Mine Y,WU J. The potential of food protein-derived anti-inflammatory peptides against various chronic inflammatory diseases[J]. J Sci Food Agric,2016,96(7):2303-2311.
[13] Manresa M C,Taylor C T. Hypoxia inducible factor(HIF)hydroxylases as regulators of intestinal epithelial barrier function[J]. Cell Mol Gas Hep,2017,3(3):303-315.
[14]郑晴,包怡敏.中药对心肌缺血再灌注损伤过程中自噬的调控[J].中国中药杂志,2017,42(15):2925-2929.
[15]涂玥,孙伟,陈涤平,等.自噬调控肾脏衰老的分子机制及中药的干预作用[J].中国中药杂志,2016,41(21):3914-3918.
[16] Serbati N,Senhaji N,Diakite B,et al. IL23R and ATG16L1 variants in moroccan patients with inflammatory bowel disease[J]. BMC Res Notes,2014,26(7):570.
[17] Baskaran K, Pugazhendhi S, Ramakrishna B S.Association of IRGM gene mutations with inflammatory bowel disease in the Indian population[J]. PLo S One,2014,9(9):e106863.
[18] Paul S,Kashyap A K,JIA W,et al. Selective autophagy of the adaptor protein Bcl10 modulates T cell receptor activation of NF-κB[J]. Immunity,2012,36(6):947-958.
[19] Castillo F,Dekonenko A,Arko-Mensah J,et al.Autophagy protects against active tuberculosis by suppressing bacterial burden and inflammation[J]. Proc Natl Acad Sci USA,2012,109(46):3168-3176.
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |