扶正解毒颗粒对猪瘟疫苗免疫效果的影响研究

发布时间：2018-07-10 20:10

本文选题：扶正解毒颗粒 + 猪瘟疫苗　；参考：《华南农业大学》2016年硕士论文

【摘要】：猪瘟是危害我国养猪业的主要传染性疾病之一,目前主要应用疫苗进行预防。由于疫苗质量参差不齐以及猪免疫低下等原因,常常产生免疫失败,导致巨大的经济损失。许多中药具有抗病毒和增强免疫的作用,且应用安全,与疫苗联合使用可增加疫苗免疫效果。本实验对主要成分为黄芪、板蓝根、淫羊藿的扶正解毒颗粒进行研究,比较了不同剂量的扶正解毒颗粒对猪瘟疫苗免疫效果的影响。实验设置了6个组,分别为空白组、免疫不给药组、黄芪多糖组(0.5g/Kg.b.w)和扶正解毒颗粒低(0.2g/Kg.b.w)、中(0.4g/Kg.b.w)、高(0.8g/Kg.b.w)剂量组。首先,检测了6组免疫后第0、10、20、30、40天猪的血常规指标(RBC、Hb)和血生化指标(AST、ALT)。其次,检测了6组免疫后第0、7、14、24、34、44、54、69、79天时猪外周血液中CFSV抗体含量,以及第0、10、20、30、40天时IgG、IgM抗体含量,并采用流式细胞术检测了嗜中性粒细胞吞噬率,以此来评价扶正解毒颗粒对猪瘟疫苗免疫后猪体液免疫应答的影响;接着,采用CD3~+、CD4~+、CD8~+荧光标记抗体结合流式细胞仪检测了猪瘟疫苗免疫后第10、20、30、40天时各组猪外周血中CD3~+、CD4~+、CD8~+T细胞的比例,比较了各组CD4~+/CD8~+比值,并以CCK-8法检测了由ConA刺激的外周血淋巴细胞增值率,以此来评价扶正解毒颗粒的使用对猪瘟疫苗免疫后猪细胞免疫应答的影响;最后,检测了猪瘟疫苗免疫后第10、20、30、40天时各组猪外周血中SOD的活性,以此来检测扶正解毒颗粒的使用对猪瘟疫苗免疫后猪体内SOD活性的影响。血常规和血生化指标检测结果显示,空白组、免疫不给药组和各给药组猪在第0、10、20、30、40天的RBC、Hb、AST、ALT结果均无显著差异,表明扶正解毒颗粒的使用对猪的健康没有不良影响。猪瘟抗体检测结果显示,注射猪瘟疫苗后第54天各组猪的CSFV抗体阻断率达到峰值;低、中、高剂量扶正解毒颗粒均能增加猪外周血中CSFV抗体含量;其中中剂量组效果最明显,该剂量组在免疫后第14天中剂量扶正解毒颗粒与猪瘟疫苗免疫联合应用组猪的CFSV抗体的阻断率就以达到50%以上;低、高剂量扶正解毒颗粒和黄芪同样能增加CFSV抗体的阻断率,但除第54天外,其余时间检测的CSFV抗体阻断率与免疫不给药组无显著差异。外周血中IgG和IgM检测结果显示,猪瘟疫苗免疫后第20天IgM的含量便迅速增加,早于IgG的出现;IgG抗体在第40天才检测到显著的增加;低、中、高剂量扶正解毒颗粒均能增加猪瘟疫苗免疫后猪外周血中IgG和IgM的含量;其中,高剂量组的效果高于中剂量组和黄芪组,但是差异不显著(p0.05)。低、中、高剂量扶正解毒颗粒对嗜中性粒细胞吞噬功能无增强作用。T淋巴细胞亚群检测结果显示,免疫后第10~20天,各组T细胞亚群比例均无显著变化,亦无显著差异(p0.05);在第30~40天,中、高剂量组能显著增加猪瘟疫苗免疫后猪外周血中CD3~+、CD4~+、CD8~+细胞比例(与免疫不给药组差异显著,p0.05),且能保持CD4~+/CD8~+比值的稳定;淋巴细胞增殖率检测结果显示,低、中、高剂量的扶正解毒颗粒均能增加疫苗免疫后外周血淋巴细胞的ConA刺激指数,且增加的强弱与实验中用药的浓度成正比。超氧化物歧化酶(SOD)活力检测结果显示,中、高剂量扶正解毒颗粒能显著增加猪瘟疫苗免疫后猪外周血中的SOD活性(与免疫不给药组差异显著,p0.05),中高剂量组之间无显著差异(p0.05)。本研究结果证明,中、高剂量的扶正解毒颗粒能通过显著增强猪瘟疫苗免疫后猪的体液免疫应答反应、细胞免疫应答反应来增强猪瘟疫苗的免疫效果,同时还能增强猪瘟疫苗免疫后外周血中SOD的活性。由于中、高剂量的扶正解毒颗粒对各指标的影响差异均不显著,临床应用时推荐使用中剂量,以节约经济成本。本实验为扶正解毒颗粒在临床上与猪瘟疫苗免疫的联合应用提供了依据。
[Abstract]:Swine fever is one of the major infectious diseases that harm the pig industry in China. At present, the main application of vaccine is to prevent it. Because of the uneven quality of the vaccine and the low immunity of pigs, it often produces immune failure and causes huge economic loss. Many Chinese medicines have the effect of antiviral and immune enhancement, and the application is safe and combined with vaccines. The main components of this experiment were Astragalus membranaceus, Radix Isatidis and Herba Epimedium. The effects of different doses of Fuzheng Jiedu Granule on the immune effect of swine fever vaccine were compared. 6 groups were set up in the experiment, which were blank group, immune non drug group, astragalus polysaccharide group (0.5g/Kg.b.w) and positive solution. Low (0.2g/Kg.b.w), medium (0.4g/Kg.b.w), and high (0.8g/Kg.b.w) dose groups. First, the blood routine indexes (RBC, Hb) and blood biochemical indexes (AST, ALT) of 6 groups of pigs after immunization were detected. Secondly, the content of CFSV antibodies in the peripheral blood of the 6 groups of pigs was detected and the 0,10,20,30,40 days were detected. IgG, IgM antibody content, and the use of flow cytometry to detect neutrophils phagocytosis rate, in order to evaluate the effect of Fuzheng Jiedu Granule on pig's humoral immune response after swine fever vaccine. Then, CD3~+, CD4~+, CD8~+ fluorescent labeling antibody combined with flow cytometry was used to detect the swine fever vaccine on 10,20,30,40 days after the immunization of each group of pigs. The ratio of CD3~+, CD4~+ and CD8~+T cells in peripheral blood was compared, and the ratio of CD4~+/CD8~+ in each group was compared, and the increment rate of peripheral blood lymphocyte stimulated by ConA was detected by CCK-8, in order to evaluate the effect of the use of Fuzheng Jiedu Granule on the immune response of pigs after immunization with swine fever vaccine. Finally, it detected the 10,20,30,40 after the immunization of the swine fever vaccine. SOD activity in the peripheral blood of all the pigs at the time of the day was used to detect the effect of the use of Fuzheng Jiedu Granule on the activity of SOD in pigs after the swine fever vaccine. The results of blood routine and blood biochemical indexes showed that there was no significant difference between the blank group, the immune non drug group and the pig group on the RBC, Hb, AST, and ALT on the day 0,10,20,30,40. The use of positive detoxification particles had no adverse effects on the health of pigs. The results of swine fever antibody test showed that the CSFV antibody blocking rate of the pigs reached the peak at fifty-fourth days after the injection of swine fever vaccine. Low, medium and high dose Fuzheng Jiedu granules could increase the content of CSFV antibody in the peripheral blood of pigs; the middle and middle dose group was most effective, and the dose group was immune after immunization. In the fourteenth days, the blocking rate of CFSV antibody in the combined application group of Fuzheng Jiedu Granule and swine fever vaccine was more than 50%, and the high dose of Fuzheng Jiedu Granule and Astragalus could also increase the blocking rate of CFSV antibody, but there was no significant difference between the anti body blocking rate of CSFV and the immunization group except fifty-fourth days except for the other time. The results of IgG and IgM detection in the peripheral blood showed that the content of IgM increased rapidly in twentieth days after the immunization of the swine fever vaccine, as early as the appearance of IgG, and the IgG antibody was significantly increased in fortieth days. The low, middle and high dose Fuzheng Jiedu granules could increase the content of IgG and IgM in the peripheral blood of swine fever vaccine, and the effect of the high dose group was higher than that of the high dose group. In the middle dose group and the Astragalus group, the difference was not significant (P0.05). Low, middle, and high dose Fuzheng Jiedu granules did not enhance the phagocytic function of neutrophils, and the results of.T lymphocyte subgroup detection showed that there was no significant change in the proportion of T cells in each group on day 10~20 after immunization, and there was no significant difference (P0.05) on the day of 30~40, in the high dose group. The proportion of CD3~+, CD4~+, CD8~+ cells in the peripheral blood of swine fever vaccine can be significantly increased (significant difference with that of the immune group, P0.05), and it can maintain the stability of the CD4~+/CD8~+ ratio, and the lymphocyte proliferation rate detection results show that the low, medium and high dose of Fuzheng Jiedu granules can increase the ConA spines of peripheral blood lymphocytes after immunization. The results of superoxide dismutase (SOD) activity detection showed that the high dose of Fuzheng Jiedu Granule could significantly increase the SOD activity in the peripheral blood of swine fever after swine fever vaccine (P0.05), and there was no significant difference between the middle and high dose groups (P0.05). The results show that the high dose of Fuzheng Jiedu Granule can enhance the humoral immune response of swine by significantly enhancing the swine fever vaccine and enhance the immune effect of the swine fever vaccine, and also enhance the viability of SOD in the peripheral blood of the swine fever vaccine. The difference of the effect of the index is not significant. The medium dose is recommended in clinical application in order to save the economic cost. This experiment provides the basis for the combined application of Fuzheng Jiedu Granule in clinical immunization with the swine fever vaccine.
【学位授予单位】：华南农业大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S858.28

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