“紫黄散”抗病毒作用及对鸡免疫功能影响的研究

发布时间：2018-04-29 21:48

本文选题：“紫黄散” + NDV　；参考：《河北科技师范学院》2015年硕士论文

【摘要】：为了研究中药复方“紫黄散”抗病毒能力以及对鸡免疫功能的影响,本课题通过检测“紫黄散”对新城疫病毒(NDV)感染鸡胚成纤维细胞(CEF)能力的影响以及其对免疫疫苗后鸡机体内抗体效价的影响,观察“紫黄散”的抗病毒能力;通过免疫器官指数的变化、免疫球蛋白含量的测定、免疫器官及粘膜免疫组织淋巴细胞数量的变化,讨论“紫黄散”对鸡机体免疫功能的影响。本研究中,我们以5000μg/mL为中药复方“紫黄散”起始作用浓度,利用2%细胞维持液(DMEM)将其倍比稀释至9.8μg/mL,共计10个浓度,分别加入到9日龄鸡胚中,对照组只加2%DMEM。72h后,利用MTT法测细胞活性以确定细胞安全浓度。随后,利用“先药后毒”、“先毒后药”、“毒药同时”三种不同的方式,将“紫黄散”和NDV加入到已形成致密单层的CEF中,MTT法观察不同的加药方式对NDV感染CEF能力的影响,并检测最适的加药浓度。另外,我们选取健康1日龄海褐兰雏鸡120只,饲喂至8日龄时随机分为四组。其中按照A高剂量组(0.2 g/只)、B中剂量组(0.1 g/只)、C低剂量组(0.05g/只)将“紫黄散”添加入日粮中,D组设为对照组。每10天从各组随机选取5只鸡剖杀,进行NDV和传染性法氏囊病毒(IBDV)抗体效价、免疫器官指数、免疫球蛋白含量的检测,利用H.E.染色及免疫组化法观察免疫器官以及粘膜免疫组织淋巴细胞数量的变化。试验结果表明:1.“紫黄散”的最大药物安全浓度为625μg/mL。感染NDV的CEF中加入“紫黄散”对细胞均具有不同程度的保护作用,其中先加入“紫黄散”后加入NDV时,细胞保护率最高,以625μg/mL尤为显著。2.三种剂量的“紫黄散”对免疫NDV和IBDV疫苗后鸡机体内抗体效价均有显著的提高作用。3.三种剂量的“紫黄散”均对免疫器官指数以及鸡机体内免疫球蛋白含量有一定促进作用,中剂量组效果最好。4.三种剂量的“紫黄散”均能促进鸡机体主要免疫器官以及粘膜免疫组织内淋巴细胞增殖。其中高剂量组在短时间内即可显著促进淋巴细胞的增殖,中剂量组则于后期发挥明显的促增殖作用。综上所述:“紫黄散”在625μg/mL时是最大安全浓度,其在该浓度下抵抗NDV感染CEF能力最强;0.2 g/只的高剂量“紫黄散”能够在短时间内迅速增强鸡机体的免疫力,0.1 g/只的中剂量“紫黄散”于后期能够更为显著地提高鸡免疫功能。由此可见,在实际生产中,于日粮中添加0.1 g/只的“紫黄散”更为适宜。
[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.
【学位授予单位】：河北科技师范学院
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S853.7

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