探讨糖尿病胃轻瘫大鼠肠道菌群失衡致免疫功能失调的机制研究及半夏泻心汤的干预作用
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摘要
目的:探讨糖尿病胃轻瘫大鼠肠道菌群失衡致免疫功能失调的机制研究及半夏泻心汤的干预作用,为治疗免疫相关性疾病提供一种治疗思路,拓宽半夏泻心汤使用范围及为中药复方免疫调节剂提供实验依据。方法:将造模成功的50只大鼠随机分为模型组、二甲双胍1.80 g/kg组、半夏泻心汤10.20、5.10、2.55 g/kg组,用16sRNA检测大鼠0.1g粪便中肠道益生菌与致病菌的数量;采用化学法检测血清中D-木糖、IgG、sIgA含量;用ELISA法检测IL-6、IL-8、IL-10、TNF-α含量;采用免疫组化方法检测CD8表达分布。结果:与正常组相比,模型组大鼠肠推进率显著降低,空腹血糖、胃残留、血清D-木糖水平明显升高;肠道形态结构明显损伤,致病菌明显增多,益生菌明显减少,IgG、IL-6、IL-8、TNF-α、CD8水平明显升高,slgA、IL-10显著降低;与模型组比较,半夏泻心汤各剂量组均能改善肠道微生态环境,促进益生菌的增殖,减少致病菌水平,调节免疫蛋白IgG、slgA及CD8表达,降低致炎因子,上调抗炎因子水平。结论:糖尿病胃轻瘫可导致高血糖、胃肠动力障碍,引起肠道菌群紊乱,降低肠道菌群多样性,损伤肠黏膜免疫屏障。半夏泻心汤能改善糖尿病胃轻瘫大鼠胃肠动力障碍,降低血糖,降低肠黏膜通透性;上调抗炎因子水平,促进益生菌生成,抑制有害菌群,减少肠道菌群对肠黏膜屏障损伤,调控肠道黏膜免疫应答。
Objective: To investigate the mechanism of immune dysfunction caused by intestinal microflora imbalance in diabetic gastroparesis(DGP) rats and the interventional effects of Banxiaxiexin Decoction, to provide new therapy ideas for immune-related diseases, and to provide the experimental basis of the application of Banxiaxiexin Decoction as TCM compound immune modulatorsMethods:50 model rats were randomly divided into DGP, DGP-MET, DGP-HD, DGP-MD and DGP-LD groups. 16 sRNA was used to detect intestinal probiotics and pathogenic bacteria in 0.1 g amount of rat feces. Chemical detection was used to determine the contents of serum D-xylose, IgG and sIgA. Also levels of IL-6, IL-8, IL-10, and TNF-α were measured by ELISA. Distribution of CD8 expression was detected by immunohistochemistry. Results: Compared with the normal group, the intestinal propulsion was significantly decreased in the model group, while fasting blood glucose, gastric residue and serum D-xylose levels were significantly increased. Intestinal morphological structure was significantly damaged, pathogenic bacteria was significantly increased and probiotics was significantly reduced. Serum levels of IgG, IL-6, IL-8, TNF-α, and CD8 levels were significantly increased, and slgA and IL-10 were significantly decreased. Compared with DGP model group,all doses of Banxiaxiexin Decoction improved the intestinal micro-ecological environment, promoted the proliferation of probiotics, reduced the pathogenic bacteria level, regulated the expressions of immune proteins IgG, slgA and CD8, reduced the inflammation factors and up-regulated the anti-inflammatory factors. Conclusion: DGP can cause high blood glucose, gastrointestinal motility disorders, intestinal flora disturbance, reduce intestinal flora diversity and damage intestinal mucosal immune barrier. Banxiaxiexin Decoction can improve the gastrointestinal motility disorders of DGP rats, reduce blood glucose, reduce intestinal permeability, up-regulate the anti-inflammatory factors, promote the production of probiotics, inhibit harmful bacteria, reduce the intestinal flora Intestinal mucosal barrier injury,and regulate the intestinal mucosal immune response.
Objective: To investigate the mechanism of immune dysfunction caused by intestinal microflora imbalance in diabetic gastroparesis(DGP) rats and the interventional effects of Banxiaxiexin Decoction, to provide new therapy ideas for immune-related diseases, and to provide the experimental basis of the application of Banxiaxiexin Decoction as TCM compound immune modulatorsMethods:50 model rats were randomly divided into DGP, DGP-MET, DGP-HD, DGP-MD and DGP-LD groups. 16 sRNA was used to detect intestinal probiotics and pathogenic bacteria in 0.1 g amount of rat feces. Chemical detection was used to determine the contents of serum D-xylose, IgG and sIgA. Also levels of IL-6, IL-8, IL-10, and TNF-α were measured by ELISA. Distribution of CD8 expression was detected by immunohistochemistry. Results: Compared with the normal group, the intestinal propulsion was significantly decreased in the model group, while fasting blood glucose, gastric residue and serum D-xylose levels were significantly increased. Intestinal morphological structure was significantly damaged, pathogenic bacteria was significantly increased and probiotics was significantly reduced. Serum levels of IgG, IL-6, IL-8, TNF-α, and CD8 levels were significantly increased, and slgA and IL-10 were significantly decreased. Compared with DGP model group,all doses of Banxiaxiexin Decoction improved the intestinal micro-ecological environment, promoted the proliferation of probiotics, reduced the pathogenic bacteria level, regulated the expressions of immune proteins IgG, slgA and CD8, reduced the inflammation factors and up-regulated the anti-inflammatory factors. Conclusion: DGP can cause high blood glucose, gastrointestinal motility disorders, intestinal flora disturbance, reduce intestinal flora diversity and damage intestinal mucosal immune barrier. Banxiaxiexin Decoction can improve the gastrointestinal motility disorders of DGP rats, reduce blood glucose, reduce intestinal permeability, up-regulate the anti-inflammatory factors, promote the production of probiotics, inhibit harmful bacteria, reduce the intestinal flora Intestinal mucosal barrier injury,and regulate the intestinal mucosal immune response.
引文
[1]IDF Diabetes Atlas.Seventh Edition.Available from:http://www.idf.org /diabetesatlas.
[2]徐萌,王吉娥,陈继兰,等.半夏泻心汤对糖尿病胃轻瘫大鼠胰岛素抵抗的影响 .中医杂志,2015,56(17)∶1502～1505.
[3]Zeng YQ,Dai Z,Lu F,et al.Emodinviacolonic irrigation modulates gut microbiota and reduces uremic toxins in rats with chronic kidney disease.Oncotarget,2016,7(14)∶17468～17478.
[4]郑伟峰,张婧卓,王丹丹.中医治疗糖尿病胃轻瘫 .长春中医药大学学报,2015,31(03)∶518～519.
[5]单书健.古今名医临证金鉴.消渴卷.中国中医药出版社,2011.
[6]卢亚敏,宋光耀,李萍.不同脂肪酸饮食对糖耐量正常人群内脏脂肪素及其胰岛素抵抗的影响 .中华糖尿病杂志,2012,20(1)∶17～19.
[7]Horowil M,Donovan D,Jones KL,et al.Gastric emptying in diabetes:clinical significance and treatment.Diabet Med,2002,19(3)∶177～194.
[8]Christophe Vanormelingen,Jan Tack ChristopherN.AndrewsDiabeticgastroparesis.British Medical Bulletin,2013,105(1)∶213～230.
[9] 顾凯时,主编.糖尿病学.上海:上海科学技术出版社,2003∶502～514.
[10]Kawamoto S,Tran T H,Maruya M,et al.The inhibitory receptor PD-1 regulates IgA selection and bacterial composition in the gut.Science,2012,336(6080)∶485.
[11]张博,窦逾常,王垂杰.脾虚老龄大鼠肠黏膜sIgA、CD3、CD8水平变化及四君子汤对其的影响 .中国老年学杂志,2014,34(11)∶3063～3064.
[12]Man A L,Gicheva N,Nicoletti C.The impact of ageing on the intestinal epithelial barrier and immune system .Cellular Immunology,2014,289(1-2)∶112.
[13]王伟宁,张熙纯,刘丽.活动期溃疡性结肠炎发病机制的免疫学探讨 .中国现代医学杂志,2013,13(16)∶ 74～78.
[14]Mac Donald T T,Monteleone G,Pender SL.Recentdevelopments in the immunology of Innammatory bowel disease .Scand J Immunol,2015,51(1)∶ 2～9.
[15]Ashwood P,Harvey R V.Functional interactions between mucosal IL -1,IL -1ra and TGF -beta 1 in ulcerative colitis .Innamm Res,2014,53(2)∶ 53～59.
[16]刘青梅,韩辅,陈曦,等.肠康饮对溃疡性结肠炎大鼠血清 IL-6、IL-10 的影响 .世界中西医结合杂志,2015,3(5)∶ 266～268.
[2]徐萌,王吉娥,陈继兰,等.半夏泻心汤对糖尿病胃轻瘫大鼠胰岛素抵抗的影响 .中医杂志,2015,56(17)∶1502～1505.
[3]Zeng YQ,Dai Z,Lu F,et al.Emodinviacolonic irrigation modulates gut microbiota and reduces uremic toxins in rats with chronic kidney disease.Oncotarget,2016,7(14)∶17468～17478.
[4]郑伟峰,张婧卓,王丹丹.中医治疗糖尿病胃轻瘫 .长春中医药大学学报,2015,31(03)∶518～519.
[5]单书健.古今名医临证金鉴.消渴卷.中国中医药出版社,2011.
[6]卢亚敏,宋光耀,李萍.不同脂肪酸饮食对糖耐量正常人群内脏脂肪素及其胰岛素抵抗的影响 .中华糖尿病杂志,2012,20(1)∶17～19.
[7]Horowil M,Donovan D,Jones KL,et al.Gastric emptying in diabetes:clinical significance and treatment.Diabet Med,2002,19(3)∶177～194.
[8]Christophe Vanormelingen,Jan Tack ChristopherN.AndrewsDiabeticgastroparesis.British Medical Bulletin,2013,105(1)∶213～230.
[9] 顾凯时,主编.糖尿病学.上海:上海科学技术出版社,2003∶502～514.
[10]Kawamoto S,Tran T H,Maruya M,et al.The inhibitory receptor PD-1 regulates IgA selection and bacterial composition in the gut.Science,2012,336(6080)∶485.
[11]张博,窦逾常,王垂杰.脾虚老龄大鼠肠黏膜sIgA、CD3、CD8水平变化及四君子汤对其的影响 .中国老年学杂志,2014,34(11)∶3063～3064.
[12]Man A L,Gicheva N,Nicoletti C.The impact of ageing on the intestinal epithelial barrier and immune system .Cellular Immunology,2014,289(1-2)∶112.
[13]王伟宁,张熙纯,刘丽.活动期溃疡性结肠炎发病机制的免疫学探讨 .中国现代医学杂志,2013,13(16)∶ 74～78.
[14]Mac Donald T T,Monteleone G,Pender SL.Recentdevelopments in the immunology of Innammatory bowel disease .Scand J Immunol,2015,51(1)∶ 2～9.
[15]Ashwood P,Harvey R V.Functional interactions between mucosal IL -1,IL -1ra and TGF -beta 1 in ulcerative colitis .Innamm Res,2014,53(2)∶ 53～59.
[16]刘青梅,韩辅,陈曦,等.肠康饮对溃疡性结肠炎大鼠血清 IL-6、IL-10 的影响 .世界中西医结合杂志,2015,3(5)∶ 266～268.