Effect and mechanism of intestinal flora disorder on the occurrence and development of SLE in mice
李亚彤 1赵珈华 1桂金秋 1刘洋 1杨恬怡 1唐小云1
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作者信息
1. 牡丹江医学院病原生物学教研室，牡丹江 157011
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摘要
目的 探讨肠道菌群紊乱对小鼠系统性红斑狼疮(systemic lupus erythematosus，SLE)发生发展的影响及机制。 方法 40只雌性美国癌症研究所(Institute of Cancer Researc，ICR)小鼠随机分为正常对照组(normal control，NC)、菌群紊乱模型组(disorder flora model，DFM)、SLE模型对照组(SLE model control group，SM)和菌群紊乱SLE组(disorder flora SLE model，DS)，每组10只。实验结束后，进行小鼠器官指数检测；HE染色观察肾脏病理改变；粪便活菌选择性培养计数分析小鼠肠道菌群的变化；酶联免疫吸附试验(enzyme linked immunosorbent assay，ELISA)检测血清中抗核抗体和抗组蛋白抗体；实时荧光定量PCR(real-time quantitative PCR，RT-qPCR)检测脾脏组织中白细胞介素(interleukin，IL)-2 mRNA的表达水平。 结果 成功建立肠道菌群紊乱小鼠模型，与DS组相比，NC组、DFM组、SM组大肠杆菌、粪肠球菌数量明显降低，双歧杆菌和乳酸杆菌数量明显升高，差异均具有统计学意义(F＝289.60、143.90、504.40、1687.00，P值均<0.05)；与DFM组/SM组相比，DS组大肠杆菌和粪肠球菌数量明显增加，双歧杆菌和乳酸杆菌数量明显减少，差异均具有统计学意义[logCFU/g：(6.94±0.01)或(6.87±0.01)比(6.96±0.01)，(5.85±0.01)或(5.77±0.01)比(5.89±0.01)，(8.91±0.01)或(8.98±0.01)比(8.73±0.02)，(8.77±0.01)或(8.92±0.00)比(8.60±0.01)，P值均<0.05]。与DS组比较，NC组、DFM组、SM组小鼠体重显著升高，脾指数、肾指数和胸腺指数显著降低，差异均有统计学意义(F＝92.02、26.99、29.43、17.06，P值均<0.05)。与NC组相比，DFM组、SM组和DS组小鼠的抗核抗体、抗双链DNA抗体水平显著增加，差异具有统计学意义(F＝332.10、1151.00，P值均<0.05)；与DFM组相比，SM组和DS组小鼠的抗双链DNA均显著增加，差异有统计学意义[pg/mL：(1.86±0.23)比(6.49±0.56)或(8.58±0.42)；ng/ mL: (7.76±0.70)比(17.66±0.14)或(18.51±0.10)，P值均<0.05]。HE染色结果显示，与NC组和DFM组相比，SM和DS组小鼠肾脏可见肾小球内细胞数量明显增多，肾间质及血管周围可见炎性细胞浸润。与NC组比较，DFM组、SM组和DS组小鼠脾脏组织IL-2 mRNA表达均显著下降，差异有统计学意义[(2.05±0.14)比(1.69±0.20)比(1.52±0.11)比(1.01±0.13)，F＝16.83，P<0.05 ]。 结论 肠道菌群紊乱能够影响相关细胞因子表达，促进SLE的炎症反应，进而加重SLE的发生发展。 Objective To investigate the effect and mechanism of intestinal flora disturbance on the development of systemic lupus erythematosus (SLE) in mice. Methods Forty female Institute of Cancer Research (ICR) mice were randomly divided into normal control group (NC), disorder flora model group (DFM), SLE model control group (SM), and disorder flora SLE model(DS), with 10 animals in each group. After the experiment, the mouse organ index was detected HE staining was used to observe renal pathological changes. Selective culture counting of fecal viable bacteria was used to analyze the changes of intestinal flora in mice. Enzyme-linked immunosorbent assay (ELISA) was used to detect antinuclear and antihistone antibodies in serum, and real-time quantitative PCR (RT-qPCR) was used to detect the expression level of interleukin (IL) -2 mRNA in spleen tissue. Results A mouse model of intestinal microbiota disorder was successfully established. Compared with the DS group, the NC group, DFM group, and SM group showed a significant decrease in the number of Escherichia coli and Enterococcus faecalis, while the number of Bifidobacterium and Lactobacillus increased significantly (F=289.60, 143.90, 504.40, 1687.00, all P values<0.05). Compared with the DFM group/SM group, the DS group showed a significant increase in the number of Escherichia coli andEnterococcus faecalis, while the number of Bifidobacterium and Lactobacillus decreased significantly[logCFU/g: (6.94±0.01) or (6.87±0.01) vs(6.96±0.01), (5.85±0.01) or (5.77±0.01) vs(5.89±0.01), (8.91±0.01) or (8.98±0.01) vs(8.73±0.02), (8.77±0.01) or (8.92±0.00) vs(8.60±0.01), all P values<0.05]. Compared with the DS group, the NC group, DFM group, and SM group showed a significant increase in body weight, while the spleen index, kidney index, and thymus index decreased significantly (F=92.02, 26.99, 29.43, 17.06, all P values<0.05). Compared with the NC group, the levels of anti-nuclear antibodies and anti-double-stranded DNA antibodies in the DFM, SM, and DS groups were significantly increased(F=332.10, 1151.00, both P values<0.05). Compared with the DFM group, the anti-double-stranded DNA levels in both SM and DS groups were significantly increased[pg/mL: (1.86±0.23) vs(6.49±0.56) or (8.58±0.42) ng/mL: (7.76±0.70) vs(17.66±0.14) or (18.51±0.10), bothP values<0.05]. The HE staining results showed that compared with the NC and DFM groups, the SM and DS groups showed a significant increase in the number of glomerular cells in the kidneys, and inflammatory cell infiltration was observed in the renal interstitium and surrounding blood vessels. Compared with the NC group, the expression of IL-2 mRNA in the spleen tissue in the DFM group, SM group, and DS group decreased significantly[(2.05±0.14) vs (1.69±0.20) vs (1.52±0.11) vs (1.01±0.13),F=16.83, P<0.05]. Conclusion The disturbance of gut microbiota can affect the expression of related cytokines, promote the inflammatory response of SLE, and further exacerbate the occurrence and development of SLE.
Abstract
Objective To investigate the effect and mechanism of intestinal flora disturbance on the development of systemic lupus erythematosus (SLE) in mice. Methods Forty female Institute of Cancer Research (ICR) mice were randomly divided into normal control group (NC), disorder flora model group (DFM), SLE model control group (SM), and disorder flora SLE model(DS), with 10 animals in each group. After the experiment, the mouse organ index was detected HE staining was used to observe renal pathological changes. Selective culture counting of fecal viable bacteria was used to analyze the changes of intestinal flora in mice. Enzyme-linked immunosorbent assay (ELISA) was used to detect antinuclear and antihistone antibodies in serum, and real-time quantitative PCR (RT-qPCR) was used to detect the expression level of interleukin (IL) -2 mRNA in spleen tissue. Results A mouse model of intestinal microbiota disorder was successfully established. Compared with the DS group, the NC group, DFM group, and SM group showed a significant decrease in the number of Escherichia coli and Enterococcus faecalis, while the number of Bifidobacterium and Lactobacillus increased significantly (F=289.60, 143.90, 504.40, 1687.00, all P values<0.05). Compared with the DFM group/SM group, the DS group showed a significant increase in the number of Escherichia coli andEnterococcus faecalis, while the number of Bifidobacterium and Lactobacillus decreased significantly[logCFU/g: (6.94±0.01) or (6.87±0.01) vs(6.96±0.01), (5.85±0.01) or (5.77±0.01) vs(5.89±0.01), (8.91±0.01) or (8.98±0.01) vs(8.73±0.02), (8.77±0.01) or (8.92±0.00) vs(8.60±0.01), all P values<0.05]. Compared with the DS group, the NC group, DFM group, and SM group showed a significant increase in body weight, while the spleen index, kidney index, and thymus index decreased significantly (F=92.02, 26.99, 29.43, 17.06, all P values<0.05). Compared with the NC group, the levels of anti-nuclear antibodies and anti-double-stranded DNA antibodies in the DFM, SM, and DS groups were significantly increased(F=332.10, 1151.00, both P values<0.05). Compared with the DFM group, the anti-double-stranded DNA levels in both SM and DS groups were significantly increased[pg/mL: (1.86±0.23) vs(6.49±0.56) or (8.58±0.42) ng/mL: (7.76±0.70) vs(17.66±0.14) or (18.51±0.10), bothP values<0.05]. The HE staining results showed that compared with the NC and DFM groups, the SM and DS groups showed a significant increase in the number of glomerular cells in the kidneys, and inflammatory cell infiltration was observed in the renal interstitium and surrounding blood vessels. Compared with the NC group, the expression of IL-2 mRNA in the spleen tissue in the DFM group, SM group, and DS group decreased significantly[(2.05±0.14) vs (1.69±0.20) vs (1.52±0.11) vs (1.01±0.13),F=16.83, P<0.05]. Conclusion The disturbance of gut microbiota can affect the expression of related cytokines, promote the inflammatory response of SLE, and further exacerbate the occurrence and development of SLE.
万方数据