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云芝糖肽对健康人外周血淋巴细胞的体外免疫调节作用
摘　要：目的：探讨云芝糖肽在体外对细胞免疫和体液免疫功能调节的可能机制。方法分离健康人外周血单个核细胞（peripheral blood mononuclear cells，PBMCs），分为PBMCs空白组、云芝糖肽20μL与PBMCs共培养组和云芝糖肽40μL与PBMCs 共培养组，3组分别于培养24 h、48 h、72 h采用流式细胞仪检测淋巴细胞亚群的比例变化及T淋巴细胞表面CD3＋／HLA－DR＋、B淋巴细胞表面CD19＋／CD69＋表达情况，ELISA方法检测上清液中IgG水平。结果云芝糖肽与PBMCs共培养组CD4^＋T细胞比例明显上升（P＜0．05），CD4^＋／CD8＋细胞比值明显增高（P＜0．05）；云芝糖肽20μL与PBMCs共培养组T细胞表面CD3^＋／HLA－DR^＋，72 h时表达水平高于24 h（P＜0．05）；B细胞表面 CD19^＋／CD69^＋的表达显示从培养24 h到48 h表达升高而在72 h表达减少的趋势；PBMCs与云芝糖肽共培养2组上清IgG水平均明显高于PBMCs空白组（P均＜0．05）。结论云芝糖肽在体外能促进CD4＋T细胞比例上升及CD8＋T细胞比例下降，提高CD4^＋／CD8^＋比值，并增加IgG分泌，具有一定的免疫调节作用。
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授予学位：硕士
学位年度：2012
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摘要:（该内容经过伪原创处理，请直接查看目录）本实验，经由过程原核表达制备绵羊α2A一AR。β2一AR第二外环肽，自动免疫年夜白鼠、小白鼠和绵羊后测定对临盆机能、屠宰目标、血清生化目标的影响。重要研讨任务以下：肾上腺素能受体肽苗的制备以DNA重组技巧构建绵羊a2A一肾上腺素能受体(α2A一AR)和β2一肾上腺素能受体(β2一AR)第二外环肽表达载体，以IPTG引诱其在年夜肠杆菌E。coli BL21(DE3)中表达，为制备便宜的肾上腺素能受体肽苗供给基本。成果注解，重组绵羊α2A一AR、β2一AR第二细胞外环份子量与预期的25。3、27。39KDa符合合。在引诱0、30、60、90和120min后重组绵羊α2A一AR第二细胞外环肽占细菌总卵白的比例分离为0%、25。0%、46。7%、61。7%和60。5%；重组绵羊β2一AR第二细胞外环肽占细菌总卵白的比例分离为0%、26。4%、40。3%、59。2%和60。0%。经亲和层析纯化后重组绵羊α2A一AR、β2一AR第二细胞外环肽分离占总卵白的97。4%、97。7%。自动免疫肾上腺素能受体对年夜白鼠发展及血液理化目标的影响选择80只体重124g阁下的雌性SD年夜白鼠，随机分红5组(每组n=16)，分离为对比组、口服β2一AR冲动剂盐酸克伦特罗3mg/kg(CL组)、自动免疫重组绵羊。2A一AR肽苗(α2A一AR组)、β2一AR肽苗(β2一AR组)和化学分解β2一AR肽苗(SP组)。免疫组在第一周首次免疫响应的抗原(70μg/只)，第三周增强免疫(35μg/只)。冲动剂组从实验开端第三周添加冲动剂(3mg/kg饲料)。对比组和冲动剂组分离在第一周和第三周免疫等体积的心理盐水。年夜白鼠自在采食和饮水，每周断料12h并称取体重，至第七周末停止。实验成果注解，重组肽苗都可安慰机体发生较高程度的抗体程度，但分解肽的抗体程度偏低。自动免疫基因工程疫苗或化学分解肽苗对年夜白鼠体重、屠宰机能均无影响(P《0。05)。CL组与对比组比拟腹脂/活重下降76。74%(P《0。05)，TG下降28。13%(P《0。05)；β2一AR组TG、TC分离比对比组降低34。15%(P《0。05)、22。94%(P《0。05)；SP组GLU比对比组降低35。45%(P《0。05)，TC降低20。59%(P《0。05)。自动免疫肾上腺素能受体对小白鼠发展及血液理化目标的影响选择100只体重22g阁下的雌性昆明小白鼠，随机分红5组(每组n=20)，分离为对比组、CL组、α2A一AR组、β2一AR组和SP组。免疫组在第一周首次免疫响应的抗原(30μg／只)，第三周增强免疫(15μg／只)。冲动剂组从实验开端第三周添加冲动剂(3mg/kg饲料)。对比组和冲动剂组分离在第一周和第三周免疫等体积的心理盐水。实验时代，年夜白鼠自在采食和饮水，每周断料12h并称取体重，至第七周末停止。实验成果注解，CL组屠宰率明显高于其它各组(p《0。05)，肠系膜脂肪占活重的百分比明显低于其它各组(p《0。05)，腿肌占活重的百分比明显高于其它各组(p《0。05)，TG比对比组下降17。69%(P》0。05)，LDL一C比对比组下降12。35%(P》0。05)；α2A一AR组、β2一AR组和SP组对年夜白鼠采食、体重和屠宰机能无影响；α2A一AR组和SP组的LDL一C分离比对比组下降19。74%(P《0。05)、12。35%(P》0。05)。自动免疫肾上腺素能受体对绵羊发展及血液理化目标的影响拔取12只体重31kg阁下雄性阿勒泰羊，随机分为对比组和β2一AR组2组。1d免疫，15d后增强免疫。自在采食和饮水，每15d称重，空肚颈静脉采血15min/次，4次。先精后粗采食1h后静脉采血15min/次，4次。90d一切植物屠宰称量胴体，血清用于测定抗体程度和生化目标。实验成果注解，β2一AR肽苗对采食、增重和饲喂后的血清生化目标均无影响；15d饲喂前β2一AR组GLU比对比组降低21。74%(P《0。05)，60d饲喂前β2一AR组LDL一C比对比组下降15。19(p《0。05)；β2一AR组屠宰率比对比组高4。60%(P《0。05)。以上成果显示：年夜白鼠、小白鼠和绵羊自动免疫肾上腺素能受体肽苗后可发生抗体，不影响年夜白鼠、小白鼠和绵羊的发展、采食量和脂肪堆积，但可进步绵羊的屠宰率。
Abstract:In this experiment, the expression of 2A in sheep was prepared by AR. Beta 2 ar second cyclic peptides, auto immune of big rats, mice and sheep determination of impact on birth function, target slaughter, serum biochemical target. Below the important research task: adrenergic receptor peptide vaccines were prepared by recombinant DNA technique to construct a sheep a2A adrenaline receptor (alpha 2a AR) and beta 2 adrenergic receptor (beta 2 AR) the second cyclic peptide expression vector, luring the eve of Enterobacter e with IPTG. BL21 coli (DE3) expression, for the preparation of the cheap to provide the basic of adrenergic receptor peptide vaccine supply. The results note that the recombinant sheep 2A a AR, beta second AR 2 cell outer ring molecular weight and the expected 25. 27, 3. 39KDa compliance. At 0, 30, 60, 90 and 120 min after recombinant ovine alpha 2a ar the second cell to lure outside loop peptides accounted for the proportion of the total bacterial protein separation of 0, 25. 46, 0. 61, 7. 7 and 60. 5; ovine beta 2 AR second extracellular loop peptide% of the total bacterial protein was separated into 0, 26 proportion. 40, 4. 59, 3. 2 and 60. 0. After purification by affinity chromatography of recombinant alpha 2A beta, a sheep AR 2 AR second extracellular loop peptide separation of total protein 97. 97, 4. 7. Epinephrine auto immune to receptors on the eve of the MICE development and blood and target 80 weight 124g your female SD big rat selection, stochastic dividends 5 groups (each group, n = 16), separation for the comparison group, oral beta 2 ar impulse agent clenbuterol hydrochloride 3mg / kg group (CL), automatic immunized with recombinant sheep. 2A a AR (2A group), beta 2 a AR peptide vaccine (beta 2 AR group) and chemical decomposition of beta 2 a AR peptide vaccine (SP group). The first immune response of the immune group was first immune response of the antigen (70 g/ only), and the third week was enhanced by immunization (35 g/ only). Third weeks from the beginning of the experiment, the impulse agent (3mg/kg) was added. The contrast group and the agonist group were isolated in the first week and the third week of the immune and other volume of the psychological saline. The rats were free to feed and drink, and the 12h was weighed every week, and stopped at the end of seventh. The results of the experiment, the recombinant peptide vaccine can stimulate the body to a higher degree of antibody level, but the degree of low antibody level of peptide. Automatic immune genetic engineering vaccine or chemical decomposition peptide vaccine has no effect on the body weight and slaughter performance (P 0. 05). CL group and the contrast group compared with abdominal fat / live weight decreased by 76. 74 (P "0. 05), TG decreased by 28. 13 (P "0. 2), the TG and TC in the AR group were decreased by 34 compared with that in the control group. 15 (P "0. 05), 22. 94 (P "0. 05); SP group GLU was lower than that of the control group by 35. 45 (P "0. 05), TC decreased by 20. 59 (P "0. 05). The effect of the automatic immune system to the development of mice and blood and the physical and chemical targets of 100 female 22G mice were randomly divided into 2 groups (n = 5), AR group, CL group, 2A group, AR group and SP group. The first immune response of the immune group was 30 g / for the first week, and third weeks (15 g / only). Third weeks from the beginning of the experiment, the impulse agent (3mg/kg) was added. The contrast group and the agonist group were isolated in the first week and the third week of the immune and other volume of the psychological saline. In the experiment, the rats were free to feed and drinking water, and the 12h was weighed every week. The slaughter rate of CL group was significantly higher than that in other groups (P 0. 05) the percentage of mesenteric fat was significantly lower than that of the other groups (P 0. 05), the percentage of leg muscle was significantly higher than that of other groups (P 0. 05), TG was decreased by 17 compared with the control group. 69 (P, 0. 05), the C LDL was decreased by 12 compared with the control group. 35 (P, 0. 05), a AR group, a AR group and a SP group had no effect on the intake, body weight and slaughter performance of rats. The 2A LDL group and AR group were decreased by 19 compared with the control group, C group and SP group. 74 (P "0. 05), 12. 35 (P, 0. 05). The effect of the automatic immune system of 31kg on the development of sheep and blood and physical and chemical targets of the 12 male Aletai sheep were randomly divided into 2 groups: the contrast group and the AR group. 1D immunization, enhanced immunity after 15d. At free feeding and drinking water, the jugular vein blood was collected from the jugular vein at 15min/ times, 4 times, times. After the first extract, the 15min/ was collected from the 1H, and the blood was collected for 4 times. 90d all plant slaughter weighing carcass, serum for the determination of antibody levels and biochemical targets. The results of the experiment showed that the serum biochemical targets of the 2 AR peptide vaccine had no effect on the feeding, weight gain and serum biochemical target. The GLU group was decreased by 21 compared with that of the AR group 15d. 74 (P "0. 05), 60d was fed with a LDL of the group, a C was decreased by 15 compared with the control group AR. 19 (P "0. 2), the slaughter rate of AR group was 4 higher than that of the control group. 60 (P "0. 05). The results showed that: the rats, mice and sheep were immunized with the auto immune adrenergic receptor peptide vaccine, which could not affect the development, feed intake and fat accumulation in rats, mice and sheep. ...
目录:摘要4-6ABSTRACT6-7第1章 文献综述12-20&&&&1.1 免疫中和12-15&&&&&&&&1.1.1 生长抑素12-14&&&&&&&&1.1.2 胆囊收缩素14&&&&&&&&1.1.3 脂肪细胞膜14-15&&&&1.2 受体免疫15-19&&&&&&&&1.2.1 受体免疫激活15-17&&&&&&&&1.2.2 受体免疫抑制17-19&&&&1.3 总结19-20第2章 重组绵羊α_(2A)-AR、β_2-AR第二外环肽在大肠杆菌中表达及肽苗的制备20-31&&&&2.1 试验材料20-21&&&&&&&&2.1.1 菌种20&&&&&&&&2.1.2 主要仪器20-21&&&&&&&&2.1.3 主要试剂21&&&&2.2 试验方法21-25&&&&&&&&2.2.1 试剂配制21-23&&&&&&&&2.2.2 重组绵羊α_(2A)-AR、β_2-AR第二外环肽在大肠杆菌中表达及纯化23-25&&&&2.3 试验结果与分析25-28&&&&&&&&2.3.1 重组绵羊α_(2A)-AR第二外环肽的诱导表达25-26&&&&&&&&2.3.2 重组绵羊β_2-AR第二外环肽的诱导表达26-27&&&&&&&&2.3.3 重组绵羊α_(2A)-AR、β_2-AR第二外环肽纯化27-28&&&&2.4 讨论28-30&&&&2.5 小结30-31第3章 主动免疫肾上腺素能受体肽苗对大白鼠、小白鼠和绵羊生长及血清生化指标的影响31-53&&&&3.1 试验材料与方法31-36&&&&&&&&3.1.1 试验动物31&&&&&&&&3.1.2 主要仪器31-32&&&&&&&&3.1.3 主要试剂32-33&&&&&&&&3.1.4 大白鼠和小白鼠免疫肾上腺素能受体肽苗33-34&&&&&&&&3.1.5 绵羊免疫重组绵羊β_2-AR第二外环肽34-35&&&&&&&&3.1.6 样品测定35-36&&&&3.2 试验结果与分析36-49&&&&&&&&3.2.1 主动免疫肾上腺素能受体肽苗对大白鼠的影响36-40&&&&&&&&3.2.2 主动免疫肾上腺素能受体肽苗对小白鼠的影响40-43&&&&&&&&3.2.3 主动免疫肾上腺素能受体肽苗对阿勒泰羊的影响43-49&&&&3.3 讨论49-52&&&&&&&&3.3.1 主动免疫肾上腺素能受体肽苗对大白鼠、小白鼠及阿勒泰羊血清抗体水平和生化指标的影响49-50&&&&&&&&3.3.2 主动免疫肾上腺素能受体肽苗对大白鼠、小白鼠及绵羊生长性能和屠宰的影响50-52&&&&3.4 小结52-53第4章 全文总结论53-54参考文献54-60致谢60-61作者简介61
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[1].王慧宇. [D]. 上海交通大学.
2014[2].张晗. [D]. 上海交通大学.
2014[3].黄晓婉. [D]. 上海交通大学.
2014[4].王昒展. [D]. 东南大学.
2013[5].徐鑫. [D]. 江苏大学.
2014[6].王欢. [D]. 江苏大学.
2014[7].何舟. [D]. 大连海洋大学.
2014[8].刘志强. [D]. 大连海洋大学.
2014[9].周元雪. [D]. 大连海洋大学.
2014[10].苏虹. [D]. 南昌大学.羊口疮怎么治疗――羊血清免疫肽
羊口疮怎么治疗――羊血清免疫肽
基本信息：男&&18岁
发病时间：不清楚
病情描述及疑问：羊口疮怎么治疗――羊血清免疫肽补充说明：之前用过香港龙达的羊血清免疫肽，效果挺好的。
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建议：你好，根据描述的病情分析属于不确切的，也是啊可能体质的，建议在医生指导下使用。
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