本文選題：“紫黃散” + NDV�。� 參考：《河北科技師范學(xué)院》2015年碩士論文

【摘要】：為了研究中藥復(fù)方“紫黃散”抗病毒能力以及對雞免疫功能的影響,本課題通過檢測“紫黃散”對新城疫病毒(NDV)感染雞胚成纖維細(xì)胞(CEF)能力的影響以及其對免疫疫苗后雞機(jī)體內(nèi)抗體效價(jià)的影響,觀察“紫黃散”的抗病毒能力;通過免疫器官指數(shù)的變化、免疫球蛋白含量的測定、免疫器官及粘膜免疫組織淋巴細(xì)胞數(shù)量的變化,討論“紫黃散”對雞機(jī)體免疫功能的影響。本研究中,我們以5000μg/mL為中藥復(fù)方“紫黃散”起始作用濃度,利用2%細(xì)胞維持液(DMEM)將其倍比稀釋至9.8μg/mL,共計(jì)10個(gè)濃度,分別加入到9日齡雞胚中,對照組只加2%DMEM。72h后,利用MTT法測細(xì)胞活性以確定細(xì)胞安全濃度。隨后,利用“先藥后毒”、“先毒后藥”、“毒藥同時(shí)”三種不同的方式,將“紫黃散”和NDV加入到已形成致密單層的CEF中,MTT法觀察不同的加藥方式對NDV感染CEF能力的影響,并檢測最適的加藥濃度。另外,我們選取健康1日齡海褐蘭雛雞120只,飼喂至8日齡時(shí)隨機(jī)分為四組。其中按照A高劑量組(0.2 g/只)、B中劑量組(0.1 g/只)、C低劑量組(0.05g/只)將“紫黃散”添加入日糧中,D組設(shè)為對照組。每10天從各組隨機(jī)選取5只雞剖殺,進(jìn)行NDV和傳染性法氏囊病毒(IBDV)抗體效價(jià)、免疫器官指數(shù)、免疫球蛋白含量的檢測,利用H.E.染色及免疫組化法觀察免疫器官以及粘膜免疫組織淋巴細(xì)胞數(shù)量的變化。試驗(yàn)結(jié)果表明:1.“紫黃散”的最大藥物安全濃度為625μg/mL。感染NDV的CEF中加入“紫黃散”對細(xì)胞均具有不同程度的保護(hù)作用,其中先加入“紫黃散”后加入NDV時(shí),細(xì)胞保護(hù)率最高,以625μg/mL尤為顯著。2.三種劑量的“紫黃散”對免疫NDV和IBDV疫苗后雞機(jī)體內(nèi)抗體效價(jià)均有顯著的提高作用。3.三種劑量的“紫黃散”均對免疫器官指數(shù)以及雞機(jī)體內(nèi)免疫球蛋白含量有一定促進(jìn)作用,中劑量組效果最好。4.三種劑量的“紫黃散”均能促進(jìn)雞機(jī)體主要免疫器官以及粘膜免疫組織內(nèi)淋巴細(xì)胞增殖。其中高劑量組在短時(shí)間內(nèi)即可顯著促進(jìn)淋巴細(xì)胞的增殖,中劑量組則于后期發(fā)揮明顯的促增殖作用。綜上所述:“紫黃散”在625μg/mL時(shí)是最大安全濃度,其在該濃度下抵抗NDV感染CEF能力最強(qiáng);0.2 g/只的高劑量“紫黃散”能夠在短時(shí)間內(nèi)迅速增強(qiáng)雞機(jī)體的免疫力,0.1 g/只的中劑量“紫黃散”于后期能夠更為顯著地提高雞免疫功能。由此可見,在實(shí)際生產(chǎn)中,于日糧中添加0.1 g/只的“紫黃散”更為適宜。
[Abstract]:In order to study the antiviral ability of traditional Chinese medicine compound "Zihuang Powder" and its effect on chicken immune function, In this study, the effect of "Zihuang Powder" on the ability of chicken embryo fibroblast (CEF) infected with NDV and its effect on antibody titer of chicken body after immunized vaccine were examined to observe the anti-virus ability of "Zihuang Powder". Through the changes of immune organ index, the content of immunoglobulin and the number of lymphocytes in immune organs and mucosal immune tissues, the effect of Zihuang Powder on the immune function of chicken was discussed. In this study, we used 5000 渭 g/mL as the initial action concentration of "Zihuang Powder" and diluted its doubling ratio to 9.8 渭 g 路mL ~ (-1) by using 2% cell maintenance solution. 10 concentrations were added to 9-day-old chicken embryo respectively. The control group was treated with 2%DMEM.72h only. The cell activity was measured by MTT method to determine the cell safe concentration. Then, using three different ways of "drug first and then poison", "poison before medicine" and "poison simultaneously", we added "Zihuang Powder" and NDV into CEF, which formed a dense monolayer, to observe the effect of different drug addition methods on the ability of NDV to infect CEF. The optimum concentration was determined. In addition, 120 1-day-old sea brown orchid chicks were randomly divided into four groups. The "Zihuang Powder" was added to the diet of Group D as the control group. Five chickens were selected randomly from each group every 10 days. The titers of NDV and infectious bursal bursal virus (IBDV) antibody, immune organ index and content of immunoglobulin were determined by H. E. The changes of lymphocyte count in immune organs and mucosal tissues were observed by staining and immunohistochemistry. The test results show that 1: 1. The maximum safe concentration of Zihuang Powder was 625 渭 g / mL. In CEF infected with NDV, "Zihuang Powder" had different protective effects on cells, especially 625 渭 g/mL, when added "Zihuang San" first and then NDV. Three doses of "Zihuang Powder" could significantly increase the titer of antibodies in chicken after immunization with NDV and IBDV vaccine. Three doses of "Zihuang Powder" can promote the immune organ index and the content of immunoglobulin in chicken, and the middle dose group has the best effect. All three doses of Zihuang Powder could promote the proliferation of lymphocytes in the main immune organs and mucosal immune tissues of chicken. The high dose group could significantly promote the proliferation of lymphocytes in a short period of time, while the middle dose group played a significant role in promoting the proliferation in the later stage. In conclusion, Zihuang Powder is the most safe concentration at 625 渭 g/mL. At this concentration, the high dose "Zihuang Powder", which has the strongest ability to resist NDV infection with CEF, can rapidly enhance the immunity of chicken in a short time. The medium dose "Zihuang Powder" of 0.1 g / per NDV can improve the immune function of chicken more significantly in the later stage. Therefore, in actual production, it is more suitable to add 0.1 g / kg "Zihuang Powder" to the diet.
【學(xué)位授予單位】：河北科技師范學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S853.7

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