發(fā)布時間：2018-07-10 20:10

  本文選題：扶正解毒顆粒 + 豬瘟疫苗　； 參考：《華南農(nóng)業(yè)大學》2016年碩士論文

【摘要】：豬瘟是危害我國養(yǎng)豬業(yè)的主要傳染性疾病之一,目前主要應(yīng)用疫苗進行預(yù)防。由于疫苗質(zhì)量參差不齊以及豬免疫低下等原因,常常產(chǎn)生免疫失敗,導致巨大的經(jīng)濟損失。許多中藥具有抗病毒和增強免疫的作用,且應(yīng)用安全,與疫苗聯(lián)合使用可增加疫苗免疫效果。本實驗對主要成分為黃芪、板藍根、淫羊藿的扶正解毒顆粒進行研究,比較了不同劑量的扶正解毒顆粒對豬瘟疫苗免疫效果的影響。實驗設(shè)置了6個組,分別為空白組、免疫不給藥組、黃芪多糖組(0.5g/Kg.b.w)和扶正解毒顆粒低(0.2g/Kg.b.w)、中(0.4g/Kg.b.w)、高(0.8g/Kg.b.w)劑量組。首先,檢測了6組免疫后第0、10、20、30、40天豬的血常規(guī)指標(RBC、Hb)和血生化指標(AST、ALT)。其次,檢測了6組免疫后第0、7、14、24、34、44、54、69、79天時豬外周血液中CFSV抗體含量,以及第0、10、20、30、40天時IgG、IgM抗體含量,并采用流式細胞術(shù)檢測了嗜中性粒細胞吞噬率,以此來評價扶正解毒顆粒對豬瘟疫苗免疫后豬體液免疫應(yīng)答的影響;接著,采用CD3~+、CD4~+、CD8~+熒光標記抗體結(jié)合流式細胞儀檢測了豬瘟疫苗免疫后第10、20、30、40天時各組豬外周血中CD3~+、CD4~+、CD8~+T細胞的比例,比較了各組CD4~+/CD8~+比值,并以CCK-8法檢測了由ConA刺激的外周血淋巴細胞增值率,以此來評價扶正解毒顆粒的使用對豬瘟疫苗免疫后豬細胞免疫應(yīng)答的影響;最后,檢測了豬瘟疫苗免疫后第10、20、30、40天時各組豬外周血中SOD的活性,以此來檢測扶正解毒顆粒的使用對豬瘟疫苗免疫后豬體內(nèi)SOD活性的影響。血常規(guī)和血生化指標檢測結(jié)果顯示,空白組、免疫不給藥組和各給藥組豬在第0、10、20、30、40天的RBC、Hb、AST、ALT結(jié)果均無顯著差異,表明扶正解毒顆粒的使用對豬的健康沒有不良影響。豬瘟抗體檢測結(jié)果顯示,注射豬瘟疫苗后第54天各組豬的CSFV抗體阻斷率達到峰值;低、中、高劑量扶正解毒顆粒均能增加豬外周血中CSFV抗體含量;其中中劑量組效果最明顯,該劑量組在免疫后第14天中劑量扶正解毒顆粒與豬瘟疫苗免疫聯(lián)合應(yīng)用組豬的CFSV抗體的阻斷率就以達到50%以上;低、高劑量扶正解毒顆粒和黃芪同樣能增加CFSV抗體的阻斷率,但除第54天外,其余時間檢測的CSFV抗體阻斷率與免疫不給藥組無顯著差異。外周血中IgG和IgM檢測結(jié)果顯示,豬瘟疫苗免疫后第20天IgM的含量便迅速增加,早于IgG的出現(xiàn);IgG抗體在第40天才檢測到顯著的增加;低、中、高劑量扶正解毒顆粒均能增加豬瘟疫苗免疫后豬外周血中IgG和IgM的含量;其中,高劑量組的效果高于中劑量組和黃芪組,但是差異不顯著(p0.05)。低、中、高劑量扶正解毒顆粒對嗜中性粒細胞吞噬功能無增強作用。T淋巴細胞亞群檢測結(jié)果顯示,免疫后第10~20天,各組T細胞亞群比例均無顯著變化,亦無顯著差異(p0.05);在第30~40天,中、高劑量組能顯著增加豬瘟疫苗免疫后豬外周血中CD3~+、CD4~+、CD8~+細胞比例(與免疫不給藥組差異顯著,p0.05),且能保持CD4~+/CD8~+比值的穩(wěn)定;淋巴細胞增殖率檢測結(jié)果顯示,低、中、高劑量的扶正解毒顆粒均能增加疫苗免疫后外周血淋巴細胞的ConA刺激指數(shù),且增加的強弱與實驗中用藥的濃度成正比。超氧化物歧化酶(SOD)活力檢測結(jié)果顯示,中、高劑量扶正解毒顆粒能顯著增加豬瘟疫苗免疫后豬外周血中的SOD活性(與免疫不給藥組差異顯著,p0.05),中高劑量組之間無顯著差異(p0.05)。本研究結(jié)果證明,中、高劑量的扶正解毒顆粒能通過顯著增強豬瘟疫苗免疫后豬的體液免疫應(yīng)答反應(yīng)、細胞免疫應(yīng)答反應(yīng)來增強豬瘟疫苗的免疫效果,同時還能增強豬瘟疫苗免疫后外周血中SOD的活性。由于中、高劑量的扶正解毒顆粒對各指標的影響差異均不顯著,臨床應(yīng)用時推薦使用中劑量,以節(jié)約經(jīng)濟成本。本實驗為扶正解毒顆粒在臨床上與豬瘟疫苗免疫的聯(lián)合應(yīng)用提供了依據(jù)。
[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.
【學位授予單位】：華南農(nóng)業(yè)大學
【學位級別】：碩士
【學位授予年份】：2016
【分類號】：S858.28

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