蒙药成份复方特润舒都乐对乳腺炎模型小鼠免疫功能的影响

发布时间：2018-04-11 08:00

本文选题：蒙药成份复方 + 乳房炎　；参考：《内蒙古农业大学》2017年硕士论文

【摘要】：奶牛乳腺炎(dairy mastitis)发病率高,且对奶牛养殖业造成严重的经济危害。其治疗的首选药物为抗生素,具有快速、显著的疗效,但抗生素长期大量使用导致病原菌耐药、乳中抗生素残留等问题,给人畜健康造成安全隐患。为此,在课题组前期研发的绿色、低毒、治疗奶牛乳房炎有效的蒙药特润舒都乐基础上,选用其抑菌、抗炎免疫有效成分替代抗生素,并以金黄色葡萄球菌和大肠杆菌混合菌液制备小鼠乳房炎病理模型,研究该成份复方对小鼠乳房炎的治疗作用及其对乳房炎模型小鼠免疫机能的影响。试验方法和结果如下:(1)组织病理切片法筛选出制备乳房炎病理模型小鼠大肠杆菌和金黄色葡萄球菌混合菌浓度均为1× 108cfu/mL。模型具有完整的病理变化过程,造模成功。(2)HE染色法制备病理组织切片,观察致病菌对小鼠乳腺组织的损伤程度,并用带测微尺的显微镜测量乳房炎模型小鼠乳腺腺泡壁厚度及间质宽度,用平板计数法检测乳腺组织匀浆中细菌的定殖数量,发现蒙药成份复方可显著降低乳腺炎模型小鼠乳腺腺泡壁厚度和菌落定殖数量(P0.05),显著减小鼠乳腺间质宽度(P0.05)。说明该蒙药成分复方具有良好的抑菌和抗炎作用,降低乳腺组织损伤。(3)用称重法测量胸腺和脾脏湿重,计算免疫脏器指数,结果提示,蒙药成份复方可以提高免疫脏器指数(P0.05)。(4)用碳阔血清法检测乳腺炎模型小鼠单核巨噬细胞吞噬活性,结果提示,蒙药成份复方显著提高乳腺炎模型小鼠单核巨噬细胞吞噬功能(P0.05)。(5)用MTT法检测乳腺炎模型小鼠NK细胞活性,结果提示,蒙药成份复方显著提高乳腺炎模型小鼠NK细胞吞噬活性(P0.05)。(6)用流式细胞术检测乳腺炎模型小鼠外周血淋巴细胞百分含量,结果蒙药成份复方显著提高乳腺炎模型小鼠CD3+T、CD4+T、CD19+B的水平,降低CD8+T水平(P0.05)。用ELISA法检测血清中免疫球蛋白含量,结果蒙药成份复方显著提高模型小鼠血清中IgG和IgA含量(P0.05)。用ELISA法检测乳腺组织匀浆中细胞因子的含量,结果蒙药成份复方显著上调 IL-10、IFN-γ含量(P0.05),下调 IL-1β、IL-2、IL-4、IL-6、IL-8、TNF-α含量(P0.05)。说明蒙药成份复方显著提高模型小鼠的细胞免疫和体液免疫水平,减少抗炎因子和提高抗炎作用。结论:蒙药特润舒都乐成份复方通过提高乳房炎模型小鼠免疫脏器指数、M0吞噬能力、NK细胞杀伤活性及稳定Thl和Th2细胞群的功能,进而增强小鼠非特性免疫及特异性免疫,以治疗E.coli和S..aureus感染所引发乳腺炎。
[Abstract]:The incidence of dairy mastitis in dairy cows is high and it causes serious economic harm to dairy cattle breeding.The first choice of antibiotics is antibiotics, which has a rapid and remarkable curative effect. However, the long-term use of antibiotics leads to the drug resistance of pathogens, antibiotic residues in milk and so on, which causes safety risks to human and animal health.Therefore, on the basis of the research and development of green, low-toxic and effective Mongolian medicine Terensudol, an effective Mongolian medicine for the treatment of dairy cow mastitis, we selected its bacteriostasis, anti-inflammatory and immune active components instead of antibiotics.The pathological model of mastitis in mice was established by mixed bacteria of Staphylococcus aureus and Escherichia coli. The therapeutic effect of this compound on mastitis in mice and the effect on immune function of mastitis model mice were studied.Methods and results of the experiment were as follows: (1) the concentration of Escherichia coli and Staphylococcus aureus in the pathological model of mastitis was 1 脳 108 cfur 路mL ~ (-1).The model had a complete pathological change process. The pathological tissue sections were prepared by the method of HE staining, and the degree of injury of the pathogenic bacteria to the mammary tissue of mice was observed.The thickness and interstitial width of mammary acinar wall in mastitis model mice were measured by microscope with micrometer, and the number of bacterial colonization in mammary tissue homogenate was detected by plate counting method.It was found that the composition of Mongolian medicine could significantly reduce the thickness of mammary acinar wall and the number of colony colonization (P0.05N) and the width of mammary stroma (P0.05) in mice with mastitis.The results showed that the compound compound of Mongolian medicine had good bacteriostatic and anti-inflammatory effects, and reduced the injury of breast tissue. The weight of thymus and spleen was measured by weighing method, and the index of immune organs was calculated. The results indicated that,The results showed that the monocyte phagocytic activity of mononuclear macrophages in mastitis model mice was detected by carbon-broad-serum method, and the results indicated that the phagocytic activity of mononuclear macrophages in mastitis model mice could be detected.The monocyte phagocytic function of mononuclear macrophages in mastitis model mice was significantly improved by Mongolian medicine composition compound. The activity of NK cells in mastitis model mice was detected by MTT method. The results indicated that the phagocytosis of mononuclear macrophages in mastitis model mice was significantly increased.The content of peripheral blood lymphocytes in mastitis model mice was detected by flow cytometry.Results the compound of Mongolian medicine could significantly increase the level of CD3 T 4 T 4 TnT CD 19 B and decrease the level of CD8 T in the mastitis model mice (P 0 05).The content of immunoglobulin in serum was detected by ELISA method. Results the content of IgG and IgA in serum of model mice was significantly increased by Mongolian medicine compound.The content of cytokines in mammary tissue homogenate was detected by ELISA method. Results the content of IFN- 纬 of IL-10 was significantly up-regulated and the content of IL-8TNF- 伪 was down-regulated by IL-1 尾 -IL-2IL-4IL-4 and IL-8TNF- 伪.The results showed that the compound of Mongolian medicine could significantly improve the cellular immunity and humoral immunity of the model mice, reduce the anti-inflammatory factor and increase the anti-inflammatory effect.Conclusion: by increasing the phagocytic activity of NK cells and stabilizing the function of Thl and Th2 cell groups in mice with mastitis model, M0 phagocytic activity and the function of Thl and Th2 cell groups can be enhanced by the Mongolian medicine Terensuldule compound, and the non-specific and specific immunity of mice can be enhanced.To treat mastitis caused by E.coli and S..aureus infections.
【学位授予单位】：内蒙古农业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S858.23

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