【作者】 陈炅然；

【导师】 杨志强；

【作者基本信息】 甘肃农业大学 ， 临床兽医学， 2008， 博士

【摘要】 为探讨藏药蕨麻的有效成分-蕨麻多糖的免疫增强和自由基药理学作用,通过体内外试验,研究了蕨麻多糖对机体的免疫功能的影响。采用流式细胞术检测蕨麻多糖对中性粒细胞呼吸爆发功能的影响。结果发现,蕨麻多糖可在中性粒细胞功能抑制时,在调理酵母多糖的刺激作用下,改善其免疫功能状态,增强中性粒细胞的呼吸爆发功能。采用环磷酰胺(Cy)作为免疫抑制剂,建立小鼠免疫抑制模型,测定和观察了小鼠脾脏指数、小鼠脾脏细胞内乳酸脱氢酶、酸性磷酸酶活性,小鼠血清细胞因子IL-10、IL-6、IFN-γ水平,小鼠脾脏中过氧化氢酶、超氧化物岐化酶、谷胱甘肽过氧化物酶活力和机体脂质过氧化程度。结果显示,蕨麻多糖可不同程度地提高免疫抑制状态下小鼠脾脏细胞乳酸脱氢酶(Lactate dehydrogenase,LDH )和酸性磷酸酶(acid phosphatase , ACP)活性,可调整IL-10和IFN-γ的平衡,升高IL-6的水平。说明蕨麻多糖通过调节中性粒细胞呼吸爆发的强度,改善因免疫抑制而引起的小鼠脾脏酸性磷酸酶水平和乳酸脱氢酶水平的下降,激活巨噬细胞的活化状态,调节促诱生细胞因子IL-10、IL-6和IFN-γ水平,拮抗因环磷酰胺引起的免疫抑制,增强机体的免疫功能。蕨麻多糖可改善因免疫抑制而引起的过氧化氢酶、超氧化物岐化酶活力的降低,通过调节抗氧化酶类的活性而去除自由基和活性氧,减轻脂质过氧化程度,分解过氧化物,阻断过氧化链,使防御体系各成分之间通过相互协同、依赖、互补作用,共同通过改善机体的氧化还原状态调节机体的免疫状态。通过蕨麻多糖不同剂量、不同途径对小鼠、兔、鸡的毒性作用和局部注射后的吸收情况,进行了蕨麻多糖的毒性试验,结果证明蕨麻多糖无毒副作用,给机体注射后可被机体缓慢吸收,造成长期持续性刺激,增强免疫效果。蕨麻多糖以不同剂量给鸡灌服或肌注,或配合新城疫弱毒疫苗给青年鸡免疫后,能升高淋巴细胞百分率,对巨噬细胞的活性具有一定的促进作用,具有促进淋巴细胞转化,升高ERFC、EAC的形成率,表明多糖对鸡的细胞免疫和体液免疫具有促进作用,能显著增强细胞和体液免疫功能,通过提高机体的非特异免疫功能而增强特异性免疫应答,达到免疫增效作用的目的。蕨麻多糖以不同途径、不同剂量免疫雏鸡,均可不同程度地提高雏鸡生长期增重;在一定程度可以提高红血球数量及血红蛋白量,从而提高机体供应养分和氧气能力,促进机体生长;对雏鸡免疫器官胸腺、法氏囊、脾脏均有不同程度的刺激作用,胸腺指数、法氏囊指数、脾脏指数都有不同程度的提高,显著促进了免疫器官的生长发育;结合新城疫疫苗,不同途径、不同剂量免疫雏鸡,可显著升高抗体水平,且抗体水平上升快,高峰值高,维持较高水平时间长,能显著增强疫苗的免疫效力。更多还原

【Abstract】 The study was carried out to investigate the the effect of immuomodulation and free radical pharmacology of potentilla anserine polysaccharide (PAP), an effective constituent from potentilla anserine , a Tibetan medicine, for its clinical application.Reactive oxygen species, stimulated by Opsonized Zymosan (OZ), was detected in neutrophilic leukocyte by flow cytometry. The result showed that PAP was able to potentialize respiratory burst produced by neutrophilic leukocyte in coordination with the stimulant under immunosuppression induced by cyclophosphamide (Cy) and induced immune response reaction in a concentration-dependent manner. The immunosuppressed model in mice was established by intraperitoneal injection with cyclophosphamide, 100 mg/kg body weight , to investigate the spleen index, the activities of lactate dehydrogenase (LDH ), acid phosphatase (ACP),catalase (CAT) , superoxide dismutase(SOD), total antioxidant capacity( TAOC ) and the extent of lipid peroxidation(LPO)in spleen , the levels of interleukin-10(IL-10), interferon(IFN) and interleukin-6(IL-6)in serum. The results showed that PAP wae able to antagonize the immunosuppression induced by Cy and increase the activities of LDH and ACP, raise levels of IL-10,IFN and IL-6 ,which suggest that PAP can potentialize immune function by regulating respiratory burst produced by neutrophilic leukocyte, activating macrophage and modulating the levels of cytokine, such as IL-10, IFN, IL-6. PAP was able to improve the activity decrease of antioxidase , such as CAT and SOD, and the extent of lipid peroxidation , extenuate oxidative damage which demonstrate that there is certain relationship between PAP immunoloregulation and free radical reaction.The toxicity test of PAP in mouse, rabbit or chicken showed that PAP has no adverse reaction and is able to enhance immune function by injection. PAP was able to reinforce immune function by enhancing function of cell mediated immunity or humoral-mediated immunity in chicken which suggest that PAP has a action on . specific immune response by potentializing nonspecific immune response. PAP was able to elevate body weight in growth phase, promote growth and development of imune organs,and raise the level of antibody combining newcastle disease vaccine in chicken.更多还原