中华消化杂志
中華消化雜誌
중화소화잡지
Chinese Journal of Digestion
2011年
9期
619-624
,共6页
叶胜青%丁在咸%张旻%陈红旗%杭晓敏%秦环龙
葉勝青%丁在鹹%張旻%陳紅旂%杭曉敏%秦環龍
협성청%정재함%장민%진홍기%항효민%진배룡
益生菌%乳杆菌属%结肠炎%白细胞介素-10%小鼠,基因敲除
益生菌%乳桿菌屬%結腸炎%白細胞介素-10%小鼠,基因敲除
익생균%유간균속%결장염%백세포개소-10%소서,기인고제
Probiotics%Lactobacillus%Colitis%Interlenkin-10%mice,knockout
目的 评估植物乳杆菌（Lactobacillus plantarum，Lp）对白细胞介素-10基因敲除(interleukin-10 knockout，IL-10-/-)小鼠结肠炎的治疗作用，并探讨其可能的作用机制。方法 8周龄雌性IL-10-/-小鼠和WT小鼠各20只，各自平均分成2组，即WT组、WT+Lp组、IL-10-/-组和IL-10-/- +Lp组。WT和1L-10-/-组予0.5 ml PBS灌胃，WT+ Lp和IL-10-/-+ Lp组予0.02g Lp(0.5 ml)灌胃，每天摄入Lp1×109菌落形成单位(CFU),持续灌胃4周后实验结束。实验开始前（0周）及开始后每隔1周收集小鼠新鲜粪便1次，直至实验结束。实验结束后将小鼠处死，记录各组小鼠体重变化，并测量其结肠长度和湿重，切取新鲜结肠组织标本做病理切片及结肠黏膜促炎因子肿瘤坏死因子(TNF)-α和干扰素-γ(IFN-γ)检测。并对小鼠新鲜粪便作选择性细菌培养，观察Lp在正常小鼠和炎症小鼠体内的定植情况及其对肠道菌群的调节作用。结果 与WT小鼠相比，IL-10-/-小鼠腹泻较重，体重亦明显下降(P＜0.05)，存在严重营养不良，而经Lp治疗后IL-10-/-小鼠腹泻得到缓解，体重亦明显增加(P＜0.05)。病理学检查显示，所有IL-10-/-小鼠皆发生肠道炎症，经Lp治疗后肠道炎症得到明显改善，黏膜溃疡、上皮增生及黏膜固有层淋巴细胞和中性粒细胞浸润明显减轻，病理学评分明显降低(P＜0.01)。IL-10-/-小鼠经Lp治疗后结肠湿重及湿重与长度比出现明显变化(P＜0.01)，结肠水肿和增厚现象得到明显改善。IL-10-/-组小鼠结肠TNF-a和IFN-γ含量分别为(377.4±84.4) μg/g和（602.6±108.1）μg/g，均较WT组明显增加[（139.2±32.7）μg/g和（173.0±52.4）μg/g，P＜0.05）]。Lp干预4周后，IL-10-/- +Lp组小鼠结肠TNF-α和IFN-γ的含量分别为(207.2±65.7) μg/g和(442.1±138.4) μg/g，均较IL-10-/-组显著降低(P＜0.05)。IL-10-/-小鼠体内肠道菌群出现紊乱。结论 Lp能有效减轻IL-10-/-小鼠肠道炎症，对结肠炎起到一定的治疗作用，且这种治疗作用与Lp调节肠道菌群及抑制促炎细胞因子的表达有关。
目的 評估植物乳桿菌（Lactobacillus plantarum，Lp）對白細胞介素-10基因敲除(interleukin-10 knockout，IL-10-/-)小鼠結腸炎的治療作用，併探討其可能的作用機製。方法 8週齡雌性IL-10-/-小鼠和WT小鼠各20隻，各自平均分成2組，即WT組、WT+Lp組、IL-10-/-組和IL-10-/- +Lp組。WT和1L-10-/-組予0.5 ml PBS灌胃，WT+ Lp和IL-10-/-+ Lp組予0.02g Lp(0.5 ml)灌胃，每天攝入Lp1×109菌落形成單位(CFU),持續灌胃4週後實驗結束。實驗開始前（0週）及開始後每隔1週收集小鼠新鮮糞便1次，直至實驗結束。實驗結束後將小鼠處死，記錄各組小鼠體重變化，併測量其結腸長度和濕重，切取新鮮結腸組織標本做病理切片及結腸黏膜促炎因子腫瘤壞死因子(TNF)-α和榦擾素-γ(IFN-γ)檢測。併對小鼠新鮮糞便作選擇性細菌培養，觀察Lp在正常小鼠和炎癥小鼠體內的定植情況及其對腸道菌群的調節作用。結果 與WT小鼠相比，IL-10-/-小鼠腹瀉較重，體重亦明顯下降(P＜0.05)，存在嚴重營養不良，而經Lp治療後IL-10-/-小鼠腹瀉得到緩解，體重亦明顯增加(P＜0.05)。病理學檢查顯示，所有IL-10-/-小鼠皆髮生腸道炎癥，經Lp治療後腸道炎癥得到明顯改善，黏膜潰瘍、上皮增生及黏膜固有層淋巴細胞和中性粒細胞浸潤明顯減輕，病理學評分明顯降低(P＜0.01)。IL-10-/-小鼠經Lp治療後結腸濕重及濕重與長度比齣現明顯變化(P＜0.01)，結腸水腫和增厚現象得到明顯改善。IL-10-/-組小鼠結腸TNF-a和IFN-γ含量分彆為(377.4±84.4) μg/g和（602.6±108.1）μg/g，均較WT組明顯增加[（139.2±32.7）μg/g和（173.0±52.4）μg/g，P＜0.05）]。Lp榦預4週後，IL-10-/- +Lp組小鼠結腸TNF-α和IFN-γ的含量分彆為(207.2±65.7) μg/g和(442.1±138.4) μg/g，均較IL-10-/-組顯著降低(P＜0.05)。IL-10-/-小鼠體內腸道菌群齣現紊亂。結論 Lp能有效減輕IL-10-/-小鼠腸道炎癥，對結腸炎起到一定的治療作用，且這種治療作用與Lp調節腸道菌群及抑製促炎細胞因子的錶達有關。
목적 평고식물유간균（Lactobacillus plantarum，Lp）대백세포개소-10기인고제(interleukin-10 knockout，IL-10-/-)소서결장염적치료작용，병탐토기가능적작용궤제。방법 8주령자성IL-10-/-소서화WT소서각20지，각자평균분성2조，즉WT조、WT+Lp조、IL-10-/-조화IL-10-/- +Lp조。WT화1L-10-/-조여0.5 ml PBS관위，WT+ Lp화IL-10-/-+ Lp조여0.02g Lp(0.5 ml)관위，매천섭입Lp1×109균락형성단위(CFU),지속관위4주후실험결속。실험개시전（0주）급개시후매격1주수집소서신선분편1차，직지실험결속。실험결속후장소서처사，기록각조소서체중변화，병측량기결장장도화습중，절취신선결장조직표본주병리절편급결장점막촉염인자종류배사인자(TNF)-α화간우소-γ(IFN-γ)검측。병대소서신선분편작선택성세균배양，관찰Lp재정상소서화염증소서체내적정식정황급기대장도균군적조절작용。결과 여WT소서상비，IL-10-/-소서복사교중，체중역명현하강(P＜0.05)，존재엄중영양불량，이경Lp치료후IL-10-/-소서복사득도완해，체중역명현증가(P＜0.05)。병이학검사현시，소유IL-10-/-소서개발생장도염증，경Lp치료후장도염증득도명현개선，점막궤양、상피증생급점막고유층림파세포화중성립세포침윤명현감경，병이학평분명현강저(P＜0.01)。IL-10-/-소서경Lp치료후결장습중급습중여장도비출현명현변화(P＜0.01)，결장수종화증후현상득도명현개선。IL-10-/-조소서결장TNF-a화IFN-γ함량분별위(377.4±84.4) μg/g화（602.6±108.1）μg/g，균교WT조명현증가[（139.2±32.7）μg/g화（173.0±52.4）μg/g，P＜0.05）]。Lp간예4주후，IL-10-/- +Lp조소서결장TNF-α화IFN-γ적함량분별위(207.2±65.7) μg/g화(442.1±138.4) μg/g，균교IL-10-/-조현저강저(P＜0.05)。IL-10-/-소서체내장도균군출현문란。결론 Lp능유효감경IL-10-/-소서장도염증，대결장염기도일정적치료작용，차저충치료작용여Lp조절장도균군급억제촉염세포인자적표체유관。
Objective To evaluate the role of Lactobacillus plantarum (LP) in the treatment of colitis in interleukin (IL)-10 knockout (IL-10-/-) mice and to explore its possible mechanisms.Methods Eight weeks old female wildtype (WT) mice and IL-10-/- mice, twenty mice of each type,were randomly assigned to four groups, WT group, WT+ LP group, IL-10-/- group and IL-I0-/- +LP group. The WT and IL-10-/- mice were gavaged with 0.5 ml saline, WT+Lp and IL-10-/- +Lp groups were gavaged with Lp 0.02 g (0.5 ml) ,took Lp 1 × 109 cfu everyday,continued for 4 weeks and then the experiment finished. The fresh mice faeces was collected once every week before (week 0) and during the experiment. The mice were executed at the end of experiments, the change of mice weight Was recorded, the length and the wet weight of colon were measured, fresh colon tissue specimens were taken for biological slices and proinflammatory cytokines TNF-a and IFN-γ were measured in colon mucosa. The fresh faeces were selectively cultured. The colonization of Lp in normal and colitis mice and its regulation role in intestinal flora were observed. Results Compared with WT mice, IL-10-/- mice demonstrated severe diarrhea, significantly decreased in body weight (P ＜0.05)and serious malnutrition. After Lp treatment, diarrhea relieved in IL-10-/- mice and the body weight increased significantly (P＜0.05). Pathological examination suggested that 100％ of IL-10-/-mice had intestinal inflammation, however after Lp treatment intestinal inflammation improved significantly. Mucosal ulcer, epithelial hyperplasia, the infiltration of neutrophils and lymphocytes in the lamina propria were also significantly reduced.The histopathological score was significantly lowered (P＜0.01). After Lp treatment, colon wet weight and the ratio of wet weight to length of IL-10-/- mice changed significantly (P＜0.01). Colon edema and thickening improved remarkably. The TNF-a and IFN-γ concentration of colon in IL-10-/- mice were 377.4±84.4 μg/g and 602.6±108.1 μg/g,which increased obviously than WT group (139.2 ± 32. 7 μg/g and 173.0± 52.4 μg/g, P＜0.05). After treated with Lp for four weeks, the TNF-α and IFN-γ concentration of colon in IL-10-/-+Lp group mice were 207.2±65.7 μg/g and 442.1 ± 138.4 μg/g, both were lower than that of IL-10-/- group mice (P＜0.05). The intestinal flora was disrupted in IL-10-/- mice. Conclusion Lp can effectively reduce intestinal inflammation in IL-10-/- mice, which take certain part in treatment in colitis. This treatment effect is associated with intestinal flora regulation and the inhibition of proinflammation cytokines expression.
