嗜酸乳杆菌治疗DSS诱导大鼠急性溃疡性结肠炎的疗效及机制研究
摘要
目的:观察嗜酸乳杆菌对DSS诱导结肠炎大鼠的治疗效果、大鼠结肠黏膜IL-1β和IL-13的表达,探讨嗜酸乳杆菌治疗UC作用机制。
方法:采用右旋葡聚糖硫酸钠(DSS)诱导的SD大鼠建立UC模型。SD雄性大鼠42只随机分为正常对照组、嗜酸乳杆菌高浓度组、嗜酸乳杆菌低浓度组、柳氮磺胺吡啶(SASP)、生理盐水(NS)对照组、模型(DSS)对照组共6组,每组7只。正常对照组自由饮水,各实验组自由饮用3%DSS水溶液,2d后改为饮用5%DSS水溶液2d,再恢复为3%DSS水溶液自由饮用,共7d建立大鼠急性UC模型,同时分别给予无菌生理盐水(NS)、嗜酸乳杆菌高剂量(1.0x1011CFU/ml)、嗜酸乳杆菌低剂量(1.0x1010CFU/ml)、SASP(67.5mg/ml)溶液灌胃治疗7d;模型组给予空针插胃。实验期间每日观察大鼠进食、饮水、活动等一般情况,称量体重,观察粪便性状及粪便隐血、便血情况。给药8d后,颈椎脱臼法处死各组大鼠,测量结肠长度,留取各组结肠标本,行HE染色。用免疫组化sp法检测在结肠黏膜上的IL-1β和IL-13表达。
结果:
1、与DSS模型组、NS组相比,其余各组治疗组大鼠一般情况好。大鼠饮用DSS后,嗜酸乳杆菌两个组、SASP组与DSS组、NS组相比,可降低DAI积分。嗜酸乳杆菌高浓度组结肠长度大于模型对照组,差异有统计学意义(P<0.05);与正常对照组相比无统计学意义(P>0.05)。2、与DSS模型组、NS组相比,嗜酸乳杆菌两个组和SASP组大鼠的结肠组织大体损伤评分和结肠组织病理学改变评分均较低,差异有统计学意义(P<0.05)。3、DSS模型组结肠黏膜IL-lβ含量高于正常组,IL-13含量低于正常组。与DSS模型组和NS组相比,嗜酸乳杆菌两个组和SASP组IL-lβ下降,IL-13呈上升趋势,有显著差异(P<0.01)。嗜酸乳杆菌组和柳氮磺胺吡啶组之间,IL-1β和IL-13的表达均无显著差异性(p>0.05)。
结论:
1、SS诱导的急性结肠炎能模拟临床UC的发作特点。嗜酸乳杆菌对大鼠急性溃疡性结肠炎有治疗作用。
2、嗜酸乳杆菌抑制UC大鼠的炎症反应可能是通过增加IL-13的表达,进而减少IL-1β的产生而实现的。
3、嗜酸乳杆菌和SASP对IL-1β和IL-13表达的影响无明显的差异性,说明嗜酸乳杆菌可以取得与抗炎药物相似的疗效。
Objective:To investigate the effects of Lactobacillus acidophilus on the expression of IL-1βand IL-13 in colonic mucosa of DSS-induced rats with Ulcerative Colitis(UC) to find its possible mechanism of treating UC.
Methods:UC rat were inducsd by dextran sulfate sodium (DSS), 42 healthy male Sprague-Daw1ey rats were randomly divided into 6 groups:normal,lactobacillus treatment(high-dose),lactobacillus treatment(1ow-dose),positive control(SASP),negative control(NS)and model groups,7 rats were involved in each group.The normal group rates were only fed with distilled water. Each experiment group freely drunk 3% DSS, for 2 days,then 3% DSS was changed to 5% DSS to feed for 2 days, last ,3% DSS for 2 days. There were 7 consecutive days and colitis was induced by SD rats.At the same time, every experimental group was orally administrated
NS(0.9%),lactobacillus(1.0x1011CFU/ml),lactobacillus(1.0x1010CFU/ml) ,SASP(67.5mg/ml),once a day individually.The body weight and the character of feces as well as occult blood were observed to know disease activity index of every mouse per day.The eight day later,all rats were sacrificed.The colon length was measured and the colony tissue was handled with HE stained.The expression of IL-1βand IL-13 in the colonic mucosa were detected by immunohistochemical staining.
Results:
1、Compare with DSS-induced rats model group and NS group general the condition of other treatment groups were better)。SD rats induced with DSS solution and Lactobacillus and SASP decreased DAI scores day by day compared with model group and NS group.Compared with model groupthe colon length of lactobacillus treatment(high-dose)group was statistically significant(P<0.05).In compared with normal group,it had no statistical significance(P>0.05).
2、Compared with model group and NS group ,gross injury score in colorectal mucosa and histopathological change of lactobacillus group and SASP group were decreased significantly, there were distinguished differences (p<0.05).
3、The IL-lβof DSS group colon mucous is higher than the normal group, the IL-13 is lower than the normal group.Compared with model group and NS group,the levels of IL-1βdecreased in lactobacillus treatmentgroup and SASP group,and the levels of IL-13 was increased in these groups(P<0.01). There's no significant difference between lactobacillus treatment group and SASP group in the levels of IL-1βand the levels of IL-13 in colorectal mucosa of the rats(P>0.05).
Conclusion:
1、The manifestations of DSS-induced acute colitis rats were similar to clinical symptoms of UC patients. Lactobacillus acidophilus have therapeutical effect to acute UC.
2、By increaseing expression of IL-13 and reducing the production of IL-1β,Lactobacillus could control inflammatory response of UC rats .
3、There's no significant difference between lactobacillus treatment groups and SASP group in the levels of IL-1βand the levels of IL-13 in colorectal mucosa of the rats,and there could be a similar results in treating UC between lactobacillus and Anti-inflammatory drugs.
方法:采用右旋葡聚糖硫酸钠(DSS)诱导的SD大鼠建立UC模型。SD雄性大鼠42只随机分为正常对照组、嗜酸乳杆菌高浓度组、嗜酸乳杆菌低浓度组、柳氮磺胺吡啶(SASP)、生理盐水(NS)对照组、模型(DSS)对照组共6组,每组7只。正常对照组自由饮水,各实验组自由饮用3%DSS水溶液,2d后改为饮用5%DSS水溶液2d,再恢复为3%DSS水溶液自由饮用,共7d建立大鼠急性UC模型,同时分别给予无菌生理盐水(NS)、嗜酸乳杆菌高剂量(1.0x1011CFU/ml)、嗜酸乳杆菌低剂量(1.0x1010CFU/ml)、SASP(67.5mg/ml)溶液灌胃治疗7d;模型组给予空针插胃。实验期间每日观察大鼠进食、饮水、活动等一般情况,称量体重,观察粪便性状及粪便隐血、便血情况。给药8d后,颈椎脱臼法处死各组大鼠,测量结肠长度,留取各组结肠标本,行HE染色。用免疫组化sp法检测在结肠黏膜上的IL-1β和IL-13表达。
结果:
1、与DSS模型组、NS组相比,其余各组治疗组大鼠一般情况好。大鼠饮用DSS后,嗜酸乳杆菌两个组、SASP组与DSS组、NS组相比,可降低DAI积分。嗜酸乳杆菌高浓度组结肠长度大于模型对照组,差异有统计学意义(P<0.05);与正常对照组相比无统计学意义(P>0.05)。2、与DSS模型组、NS组相比,嗜酸乳杆菌两个组和SASP组大鼠的结肠组织大体损伤评分和结肠组织病理学改变评分均较低,差异有统计学意义(P<0.05)。3、DSS模型组结肠黏膜IL-lβ含量高于正常组,IL-13含量低于正常组。与DSS模型组和NS组相比,嗜酸乳杆菌两个组和SASP组IL-lβ下降,IL-13呈上升趋势,有显著差异(P<0.01)。嗜酸乳杆菌组和柳氮磺胺吡啶组之间,IL-1β和IL-13的表达均无显著差异性(p>0.05)。
结论:
1、SS诱导的急性结肠炎能模拟临床UC的发作特点。嗜酸乳杆菌对大鼠急性溃疡性结肠炎有治疗作用。
2、嗜酸乳杆菌抑制UC大鼠的炎症反应可能是通过增加IL-13的表达,进而减少IL-1β的产生而实现的。
3、嗜酸乳杆菌和SASP对IL-1β和IL-13表达的影响无明显的差异性,说明嗜酸乳杆菌可以取得与抗炎药物相似的疗效。
Objective:To investigate the effects of Lactobacillus acidophilus on the expression of IL-1βand IL-13 in colonic mucosa of DSS-induced rats with Ulcerative Colitis(UC) to find its possible mechanism of treating UC.
Methods:UC rat were inducsd by dextran sulfate sodium (DSS), 42 healthy male Sprague-Daw1ey rats were randomly divided into 6 groups:normal,lactobacillus treatment(high-dose),lactobacillus treatment(1ow-dose),positive control(SASP),negative control(NS)and model groups,7 rats were involved in each group.The normal group rates were only fed with distilled water. Each experiment group freely drunk 3% DSS, for 2 days,then 3% DSS was changed to 5% DSS to feed for 2 days, last ,3% DSS for 2 days. There were 7 consecutive days and colitis was induced by SD rats.At the same time, every experimental group was orally administrated
NS(0.9%),lactobacillus(1.0x1011CFU/ml),lactobacillus(1.0x1010CFU/ml) ,SASP(67.5mg/ml),once a day individually.The body weight and the character of feces as well as occult blood were observed to know disease activity index of every mouse per day.The eight day later,all rats were sacrificed.The colon length was measured and the colony tissue was handled with HE stained.The expression of IL-1βand IL-13 in the colonic mucosa were detected by immunohistochemical staining.
Results:
1、Compare with DSS-induced rats model group and NS group general the condition of other treatment groups were better)。SD rats induced with DSS solution and Lactobacillus and SASP decreased DAI scores day by day compared with model group and NS group.Compared with model groupthe colon length of lactobacillus treatment(high-dose)group was statistically significant(P<0.05).In compared with normal group,it had no statistical significance(P>0.05).
2、Compared with model group and NS group ,gross injury score in colorectal mucosa and histopathological change of lactobacillus group and SASP group were decreased significantly, there were distinguished differences (p<0.05).
3、The IL-lβof DSS group colon mucous is higher than the normal group, the IL-13 is lower than the normal group.Compared with model group and NS group,the levels of IL-1βdecreased in lactobacillus treatmentgroup and SASP group,and the levels of IL-13 was increased in these groups(P<0.01). There's no significant difference between lactobacillus treatment group and SASP group in the levels of IL-1βand the levels of IL-13 in colorectal mucosa of the rats(P>0.05).
Conclusion:
1、The manifestations of DSS-induced acute colitis rats were similar to clinical symptoms of UC patients. Lactobacillus acidophilus have therapeutical effect to acute UC.
2、By increaseing expression of IL-13 and reducing the production of IL-1β,Lactobacillus could control inflammatory response of UC rats .
3、There's no significant difference between lactobacillus treatment groups and SASP group in the levels of IL-1βand the levels of IL-13 in colorectal mucosa of the rats,and there could be a similar results in treating UC between lactobacillus and Anti-inflammatory drugs.
引文
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3.中华医学会消化病学分会炎症性肠病协作组.对我国炎症性肠病诊断治疗规范的共识意见[J].世界华人消化杂志,2008,16(11): 1141-1143
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1.Shanahan F. Inflammatory bowel disease : immunodiagnostics ,immunotherapeutics,and ecotherapeutics[J]. Gastroenterology.2001;120(3):622—635
2.Hendrickson BA,Gokhale R,Cho JH. Clinical aspects and pathophysiology of inflammation bowel disease[J].Clin Microbiol Rev,2002,15(1):79-94
3.中华医学会消化病学分会炎症性肠病协作组.对我国炎症性肠病诊断治疗规范的共识意见[J].中华消化杂志,2007,27:545-550
4. Moum B,Ekbom A,Vatn MH,et a1.Clinical course during the 1st year after diagnosis in ulcerative colitis and Crohn’s disease.Results of a large,prospective population-based study in southeastern Norway,1990-93[J]. Scand J Gastroenterol,1997,32:1005-1012
5.Sartor RB.Therapeutic manipulation of the enteric microflorain ininflammatory bowel disease : antibiotics , probiotics , and prebiotics[J].Gastroenterology.2004;126(6):1620-1633
6. Dotan I,Rachmilewitz D.Probiotics in inflammatory bowel disease:possible mechanisms of action[J].Curr Opin in Gastroentero1.2005;21(4):426-430
7.Marteau P, Pochart P,Dore J,et a1.Comparative study of bacterial groups within the human cecal and fecal microbiota[J] . Appl Environ Microbi01.2001;67(10):4939-4942
8.Seksik P,Rigottier-Gois L,Gramet G,et a1.Alterations of the dominant faecal bacterial groups in patients with Crohn′s disease of the colon[J]. Gut,2003,52:237-242
9.Swidsinski A,Weber J,Loening-Baueke V,et a1.Spatial organization and composition of the mucosal flora in patients with inflammatory bowel disease[J].J Clin Microbiol,2005,43:3380-3389
10. Schultsz C,Van Den Berg FM,Ten Kate Fw,et a1.The intestinal mucus layer from patients with inflammatory bowel disease harbors high numbers of bacteria compared with control s [J].Gastroenterology,1999, 117:1089-1097
11.Hans W,Scholmerich J,Gross V,et a1.The role of the resident intestinal flora in acute and chronic dextran sulfate sodium-induced colitis in mice[J].Eur J Gastroenterol Hepatol,2000,12(3):267
12.Ohkusa T,Okayasu I,Ogihara T,et a1.Induction of experimental ulcerative colitis by fusobacterium varium isolated from colonic mucosa of patients with ulcerative colitis[J].Gut,2003,52:79
13.Lilly DM , Stillwell RH. . Growth—promoting factors produced by microorganisms[J].Science,1965,147:747-748
14.刘清泉.最具市场竞争力的益生菌发展现状及值得关注的几个问题[J].中国食品添加剂专家论坛,2006,6:47-60
15.刘建生,田怡,张晓红,等.溃疡性结肠炎时一氧化氮和超氧化物歧化酶变化及益生菌对其影响机制[J].中国医师进修杂志,2007,30:(6)20-22
16. Borruel N,Carol M,Casellas F,et a1.Increased mueosaltumour necrosis factor alpha production in Crohn’s disease can be downregulated ex vivo by probiotic bacteria[J].Gut,2002,51:659
17.Guslandi M,Giollo P,Testoni PA.A pilot trial of saccharomyces boulardii in ulcerative colitis [J] .Eur J Gastroenterol Hepatol,2003,15(6):697
18.Kato K, Mizuno s,Umeaaki Y,et a1.Randomized placebo- controlled trial assessing the effect of bifidobacteria-fermented milk on active ulcerative colitis[J].Aliment Pharmacol Ther,2004,20:1133
19.Zocco MA,dal Verme LZ,Cremonini F et a1.Efficacy of Lactobacillus GG in maintaining remission of ulcerative colitis [J].Aliment Pharmaeol Ther,2006,23(11):1 567-1 574
20.Reid G,Burton J.Use of Lactobacillus to prevent infection by pathogenic bacteria.Microbes Infect,2002,4:319-324
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23. Puthenedam M, Williams PH, Lakshmi BS, Balakrishnan A. Modulation of tight junction barrier function by outer membrane proteins of enteropathogenic Escherichia coli: role of F-actin and junctional adhesion molecule-1[J]. Cell Biol Int 2007; 31: 836-844
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38.Caballero-Franco C,Keller K De Simone C,et a1.The VS#3 probiotic formula induces mucin gene expression and secretion in colonic epithelial cells[J].Am J Physiol Gastrointest Liver Physiol 2007;292(1):G315-G322
39.Liu Z, Zhang P, Ma Y, Chen H, Zhou Y, Zhang M, Chu Z, Qin H. Lactobacillus plantarum prevents the development of colitis in IL-10-deficient mouse by reducing the intestinal permeability[J]. Mol Biol Rep 2011 Feb;38(2):1353-1361
40.Liu ZH, Ma YL, Shen TY, Chen HQ, Zhou YK, Zhang P, Zhang M, Chu ZX, Qin HL. Identification of DC-SIGN as the receptor during the interaction of lactobacillus plantarum CGMCC 1258 and dendritic cells[J] World Journal of Microbiology and Biotechnology, 2011:27(3): 603-611(9)
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1.Shanahan F. Inflammatory bowel disease : immunodiagnostics ,immunotherapeutics,and ecotherapeutics[J]. Gastroenterology.2001;120(3):622—635
2.Hendrickson BA,Gokhale R,Cho JH. Clinical aspects and pathophysiology of inflammation bowel disease[J].Clin Microbiol Rev,2002,15(1):79-94
3.中华医学会消化病学分会炎症性肠病协作组.对我国炎症性肠病诊断治疗规范的共识意见[J].中华消化杂志,2007,27:545-550
4. Moum B,Ekbom A,Vatn MH,et a1.Clinical course during the 1st year after diagnosis in ulcerative colitis and Crohn’s disease.Results of a large,prospective population-based study in southeastern Norway,1990-93[J]. Scand J Gastroenterol,1997,32:1005-1012
5.Sartor RB.Therapeutic manipulation of the enteric microflorain ininflammatory bowel disease : antibiotics , probiotics , and prebiotics[J].Gastroenterology.2004;126(6):1620-1633
6. Dotan I,Rachmilewitz D.Probiotics in inflammatory bowel disease:possible mechanisms of action[J].Curr Opin in Gastroentero1.2005;21(4):426-430
7.Marteau P, Pochart P,Dore J,et a1.Comparative study of bacterial groups within the human cecal and fecal microbiota[J] . Appl Environ Microbi01.2001;67(10):4939-4942
8.Seksik P,Rigottier-Gois L,Gramet G,et a1.Alterations of the dominant faecal bacterial groups in patients with Crohn′s disease of the colon[J]. Gut,2003,52:237-242
9.Swidsinski A,Weber J,Loening-Baueke V,et a1.Spatial organization and composition of the mucosal flora in patients with inflammatory bowel disease[J].J Clin Microbiol,2005,43:3380-3389
10. Schultsz C,Van Den Berg FM,Ten Kate Fw,et a1.The intestinal mucus layer from patients with inflammatory bowel disease harbors high numbers of bacteria compared with control s [J].Gastroenterology,1999, 117:1089-1097
11.Hans W,Scholmerich J,Gross V,et a1.The role of the resident intestinal flora in acute and chronic dextran sulfate sodium-induced colitis in mice[J].Eur J Gastroenterol Hepatol,2000,12(3):267
12.Ohkusa T,Okayasu I,Ogihara T,et a1.Induction of experimental ulcerative colitis by fusobacterium varium isolated from colonic mucosa of patients with ulcerative colitis[J].Gut,2003,52:79
13.Lilly DM , Stillwell RH. . Growth—promoting factors produced by microorganisms[J].Science,1965,147:747-748
14.刘清泉.最具市场竞争力的益生菌发展现状及值得关注的几个问题[J].中国食品添加剂专家论坛,2006,6:47-60
15.刘建生,田怡,张晓红,等.溃疡性结肠炎时一氧化氮和超氧化物歧化酶变化及益生菌对其影响机制[J].中国医师进修杂志,2007,30:(6)20-22
16. Borruel N,Carol M,Casellas F,et a1.Increased mueosaltumour necrosis factor alpha production in Crohn’s disease can be downregulated ex vivo by probiotic bacteria[J].Gut,2002,51:659
17.Guslandi M,Giollo P,Testoni PA.A pilot trial of saccharomyces boulardii in ulcerative colitis [J] .Eur J Gastroenterol Hepatol,2003,15(6):697
18.Kato K, Mizuno s,Umeaaki Y,et a1.Randomized placebo- controlled trial assessing the effect of bifidobacteria-fermented milk on active ulcerative colitis[J].Aliment Pharmacol Ther,2004,20:1133
19.Zocco MA,dal Verme LZ,Cremonini F et a1.Efficacy of Lactobacillus GG in maintaining remission of ulcerative colitis [J].Aliment Pharmaeol Ther,2006,23(11):1 567-1 574
20.Reid G,Burton J.Use of Lactobacillus to prevent infection by pathogenic bacteria.Microbes Infect,2002,4:319-324
21.Bernet MF,Brassart D,Neeser JR,et a1.Lactobacillus acidophilus LA1 binds to cultured human intestinal cell lines and inhibits cell attachment and cell invasion by enterovirulent bacteria[J].Gut,1994,35:483-489
22.Forster C. Tight junctions and the modulation of barrier function in disease[J]. Histochem Cell Biol 2008;130: 55-70
23. Puthenedam M, Williams PH, Lakshmi BS, Balakrishnan A. Modulation of tight junction barrier function by outer membrane proteins of enteropathogenic Escherichia coli: role of F-actin and junctional adhesion molecule-1[J]. Cell Biol Int 2007; 31: 836-844
24.Schmitz H,Barmeyer C,Fromm M,et a1.Altered tight junction structure contributes to the impaired epithelial barrier function in ulcerative colitis[J].Gastroenterology,1999,116(2):301-309
25.Qin H, Zhang Z, Hang X, et al. L. plantarum prevents enteroinvasive Escherichia coli-induced tight junction proteins changes in intestinal epithelial cells[J]. BMC Microbiol 2009;9:63
25.Resta-Lenert S,Barrett KE.Live probiotics protect intestinal epithelial cells from the effects of infection with enteroinvasive Escherichia coli(EIEC) [J].Gut,2003;52(7):988-997
26.Pena JA,Versalovic J.Lactobacillus rhamnosus GG Decreases TNF-alpha production in lipopolysaccharideactivated murine macrophages by a contact—independent mechanism[J].Cell Microbiol,2003,5:277-285
27.Fabia R,Ar.Rajab A,Johansson ML,et a1.The effect of exogenous administration of Lactobacillus reuteri R2LC and oat fiber on acetic acid-induced colitis in the rat [J].Scand J Gastroenterol,1993,28(2):155-162
28.White JS, Hoper M, Parks RW, Clements WD, Diamond T, Bengmark S.The probiotic bacterium Lactobacillus plantarum species 299 reduces intestinal permeability in experimental biliary obstruction[J]. Lett Appl Microbiol 2006; 42: 19-23
29.杨俊,张中伟,秦环龙.乳酸菌对肠上皮细胞侵袭性大肠杆菌损伤的保护作用[J].世界华人消化杂志2008; 16:3394-3399
30.王立生,潘令嘉.施理,等.双歧杆菌对裸鼠腹腔巨噬细胞产生IL-1及IL-6的影响[J].中国微生态学杂志.1998.10(3):129-131
31.Neish AS,Gewirtz AT,Zeng H,et a1.Prokaryotic regulation of epithelial responses by inhibition of IkappaB-alpha ubiquitination [J].Science,2000,289(5484):1560-1563
32.杨玉荣,郑世民,马春全,等.雏鸡服用益生素后局部粘膜免疫组织抗体生成细胞的动态变化[J].中国预防兽医学报,2004,26(5):355-358
33.霍丽娟,赵和平.益生菌对溃疡性结肠炎大鼠结肠粘膜中lL-l和TNF-a表达的影响[J].山西医科大学学报,2004,35:341-343-349
34.HUUB J M. Interaction of bifidobacterial lipoteichoic acid with human intestinal epithelial cells[J].Infect Immun,1985,47(1):332-334
35.Lievin V,Peiffer I,Hudault S,et a1.Bifidobacterium strains form resident infant human gastrointestinal microflora exert antimicrobial activity[J].Gut,2000,47(5):646-652
36.Maassen CB, van Holten-Neelen C, Balk F, den Bak-Glashouwer MJ, Leer RJ, Laman JD, Boersma WJ, Claassen E. Strain-dependent induction of cytokine profiles in the gut by orally administered Lactobacillus strains[J].Vaccine 2000; 18(22): 2613-2623
37.Sartor RB. Therapeutic manipulation of the enteric microflora in inflammatory bowel diseases: antibiotics, probiotics, and prebiotics [J].Gastroenterology,2004,126(6):1620-1633
38.Caballero-Franco C,Keller K De Simone C,et a1.The VS#3 probiotic formula induces mucin gene expression and secretion in colonic epithelial cells[J].Am J Physiol Gastrointest Liver Physiol 2007;292(1):G315-G322
39.Liu Z, Zhang P, Ma Y, Chen H, Zhou Y, Zhang M, Chu Z, Qin H. Lactobacillus plantarum prevents the development of colitis in IL-10-deficient mouse by reducing the intestinal permeability[J]. Mol Biol Rep 2011 Feb;38(2):1353-1361
40.Liu ZH, Ma YL, Shen TY, Chen HQ, Zhou YK, Zhang P, Zhang M, Chu ZX, Qin HL. Identification of DC-SIGN as the receptor during the interaction of lactobacillus plantarum CGMCC 1258 and dendritic cells[J] World Journal of Microbiology and Biotechnology, 2011:27(3): 603-611(9)