【作者】 刘迎；

【导师】 钟秀会； 史万玉；

【作者基本信息】 河北农业大学 ， 临床兽医， 2006， 硕士

【摘要】 “促免散”是由中药黄芪和淫羊藿按照一定比例配伍而成的一个具有免疫调节作用的纯中药复方。通过对超微粉和普通粗粉的对比试验,来探讨“促免散”的免疫促进作用以及该方超微粉促免功效优于传统粗粉的机理。应用激光粒度仪和扫描电镜对微粉的粒度和破壁情况进行了观测,测定了“促免散”微粉的堆密度及流动性,并与普通粗粉进行了比较。此外,还应用称重法、蒽酮-硫酸法以及反相高效液相色谱法分别比较分析了微粉与粗粉的总浸出物、有效成分多糖以及淫羊藿苷溶出率的差异。结果发现黄芪和淫羊藿经超微粉碎后,粉体的粒度分布中心D₅₀分别为6.82μm和8.40μm,经电镜观察均难以看到完整的细胞形态,只能看到细胞碎片,破壁率远远大于普通粗粉,其混合的均匀度显著提高,而普通粗粉在显微镜下,组织块清晰可见,且形状不规则,粒径分布不均匀,组织细胞仍然完整;超微粉碎后,黄芪和淫羊藿的堆密度和流动性显著增强;通过对淫羊藿有效成分淫羊藿苷的色谱分析,其中淫羊藿超微粉的淫羊藿苷体外溶出量较淫羊藿粗粉中提高了18.31%;同时黄芪、淫羊藿超微粉多糖的提取率分别较黄芪、淫羊藿粗粉提高了117.13%,315.15%。在各种提取条件下,黄芪、淫羊藿超微粉的总浸出物分别较相同条件下的普通粗粉组提高了15.41%～40.59%,4.50%～66.97%。本试验将98只昆系小鼠随机平均分为2组,为正常组和免疫抑制模型组,每组各设粗粉组、微粉组和生理盐水对照组,粗粉和微粉各设低、中、高三个剂量组。低、中、高剂量组小鼠分别灌服1∶1的“促免散”粗粉、微粉煎液0.2mL/只/天、0.4mL/只/天和0.6mL/只/天,生理盐水对照组小鼠灌服生理盐水0.4mL/只/天,共灌药6天。免疫抑制小鼠用氢化可的松腹腔注射造模。各组连续灌服6d后,分别检测小鼠的迟发型变态反应（DTH）以及外周血T细胞亚群（CD4⁺/CD8⁺）和血清TNF-α含量的变化情况。结果表明,“促免散”对正常和免疫抑制小鼠足趾变态反应、外周血CD4⁺/CD8⁺T淋巴细胞亚群的比例以及正常小鼠的血清TNF-α的含量均有一定的促进和提高作用,但与粉体的粒度和剂量有关,微粉低剂量即可达到与粗粉中高剂量相当的效果。提示“促免散”对小鼠细胞免疫功能具有一定的增强作用,且微粉优于粗粉。更多还原

【Abstract】 "Cumiansan" is a Chinese herbal medicine prescription, composed of Astragalus membranaceus and Epimedium brevicornum in proper proportion which has the immunoenhancement function. Through the contrast experiment of ultra-fine powder and commonly cribble, the immunological enhancement function of the herbal formula were investigated, as well as the mechanism that immune effect of ultra-fine powder was better than that of commonly cribble. The granularity and broken cellular wall of ultra-fine powder were studied by JL9200 laser analytical instrument of granularity and scanning electron microscopy(SEM), as well as the bulk density and liquidity of "Cumiansan" ultra-fine powder were measured, and compared to the commonly cribble. In addition, analysis was carried out in the differences of the water extract, effective component polysaccharide and the icariin distilled rate from the commonly cribble and the ultra-fine powder by weighing, anthrone-sulfuric acid method and RP-HPLC. The results showed that the D50 of Astragalus membranaceus and Epimedium brevicornum ultra-fine powder were 6.82μm and 8.40μm separately. The broken cellular walls of two ultra-fine herbal powder were bigger than those of the commonly cribble, and intact cells were not visible under SEM, and it improved the mixed uniformity. On the contrary, organization blocks of the commonly cribble were clearly discernible, also the shape and granularity were irregular, and the organization cells were still complete. The bulk density and liquidity of Astragalus membranaceus and Epimedium brevicornum increased significantly after ultra-fine Smashing. The component icariin extraction content of the ultra-fine powder was enhanced by 18.31% compared with the cribble by the chromatograph analysis of icariin. At the same time, Astragalus membranaceus polysaccharide and Epimedium brevicornum polysaccharide content of the ultra-fine powder were enhanced by 117.13% and 315.15% compared with the cribble. Under every condition, The Astragalus membranaceus and Epimedium brevicornum aqueous extract extraction rate of the ultra-fine powder were enhanced by 15.41%~40.59%, 4.50%~66.97%. In the experiment, 98 KunMing mice were randomly divided into 2 groups, which were normal mice group and immunosuppressive mice group. Each group included: commonly cribble group, ultra-fine powder group andnormal saline (NS) group. In commonly cribble and ultra-fine powder group, heavy, middle and slight dose groups were included. Every heavy, middle and slight dose groups mice were administered with the pulyis by gavage every day at 0.6mL, 0.4mL, 0.2mL and every NS group mouse were ig 0.4mL NS every day. The immunosuppressive mice model were established by intraperitoneal injection of hydrocortisone (Hy). All groups of mice were administered with the herbal pulyis by gavage for six days consecutively. The thickness of foot-plate in DTH, CD4+/CD8+T cell ratio and the concentration of TNF-a in serum of each group were observed. The results were listed as follows: "Cumiansan" had certain promotion and enhancement to the thickness of foot-plate in DTH, CD4+/CD8+T cell ratio in peripheral blood of the normal and the immunosuppressive mice as well as the normal mice’s blood serum TNF-a content, the ultra-fine powder remarkably surpassed the commonly cribble. These indicated that "Cumiansan" had certain enhancement to the cellular immunity function of the mice.更多还原