【作者】 杨树东；

【导师】 包海鹰；

【作者基本信息】 吉林农业大学 ， 中药学， 2007， 硕士

【摘要】 胶陀螺[Bulgaria inquinans(Pers．)Fr．]属于胶陀螺科的子囊菌。是一种特殊的很有开发潜力的食药兼用菌，具有抗肿瘤、光敏活性、降低血瘀大鼠的红细胞沉降率、抗突变等作用，营养成分的含量也很高。主要分布于我国东北地区，并且在煤矿中不见光的地方常出现此菌。现在国内已有企业将胶陀螺开发成治疗银屑病的新药，日本也在中国大量收购此菌，对袋料栽培和大面积推广需要进一步研究，为今后人工栽培及原料的获得提供依据。随着光动力疗法的深入研究，尤其应明确光敏活性成分及其中毒机理，制定相应的药材质量标准。本文对胶陀螺生药学方面的研究主要表现在性状特征、显微特征、理化特征及分子鉴定方面。子实体初期呈黄褐色球形，并带有黄棕色麻点，顶端逐渐开裂，产生一至多个不等的裂口，裂口不断扩大，最后整个子实体表面呈黑色皱缩。子囊棒形，外囊盘被由圆形细胞组成，圆形细胞外侧生绒毛，绒毛细胞内含类似载色体的物质；中囊盘被由埋在胶质中的交错菌丝组成。采用子囊孢子弹射分离法所得菌丝有橘黄色菌丝和长有棒状分生孢子的无色有隔菌丝；采用组织分离法所得菌丝无色，其中生有黑褐色或黄棕色的圆形细胞，呈念珠状。胶陀螺子实体DNA序列与组织分离菌种经液体培养获得的菌丝体DNA序列比对，同源性为99.65％，胶陀螺子实体的DNA序列与GenBank数据库中AY789345比对，其同源性为99.80％，说明组织分离得到的菌种稳定可靠，是胶陀螺纯菌种。本文通过对胶陀螺子实体和发酵过的子实体的提取分离，共得到5个化合物，1个具光敏活性的组分，鉴定其中4个化合物分别为丙二酸，十六酸，4，8-十九碳二烯酸，2，3-邻萘-4-(1-羟基-正丁基)-5-戊内酯，其中后3个化合物为首次从胶陀螺中得到，2，3-邻萘-4-(1-羟基-正丁基)-5-戊内酯为新化合物，光敏活性组分初步断定为苝醌类成分。从石油醚萃取层中得到的Q组分采用GC-MS联用对其分析，主要检索到22个化合物。从氯仿萃取层中得到的G组分采用GC-MS联用对其分析，主要检索到7个化合物。它们主要是一类极性较小的脂肪族烃及脂肪酸。通过光敏活性实验表明，胶陀螺丙酮粗提物、丙酮粗提物中萃取得到的氯仿萃取物及从氯仿萃取物中分离得到的Y4组分和Y5组分具有光敏活性，丙酮粗提物1000mg／kg给药的光照组耳指数有显著差异，2000mg／kg的剂量给药的光照组耳指数和皮指数均有有极显著差异。氯仿萃取组分以260mg／kg和520mg／kg的剂量给药的光照组耳指数和皮指数均有极显著差异。Y4组分以200mg／kg给药的光照组耳指数有显著差异，Y5组分以200mg／kg和100mg／kg的剂量给药的光照组耳指数均有极显著差异，Y4组分以200mg／kg及Y5组分以200mg／kg和100mg／kg的剂量给药的光照组皮指数均有极显著差异。光敏活性与剂量间存在一定的量效关系，并且其光敏毒性的发作也有一定的潜伏期。大鼠以丙酮提取物1g／kg连续给药五天，每天照射光两小时，第四天大鼠耳朵开始溃烂，爪子也变得比较紫，第六天耳朵溃烂掉了。大鼠实验也验证了胶陀螺具有光敏活性，并且对见光敏感的部位有很大的损伤。更多还原

【Abstract】 Bulgaria inquinans is a kind of ascomycete that Belongs to Bulgariaceae. It is a kind of special development potential fungi that to be used of food and medicine concurrently, it has the anti-tumor action, the photoactive activity, reduceing red blood cell rate of sedimentation of blood stasis rat, anti-sudden change and so on. The ingredient content of nutrition is very high. It mainly distributes in northeast area of our country, and this fungi often appear in the aphotic place of the coal mine. Now domestic enterprise has developed new medicine with Bulgaria inquinans treating psoriasis, this fungi also were massively purchased by Japanese from China, the bag material cultivation and the big area generalization needs to further investigation, that will provides the evidence for factitial cultivation and to gain raw material. Along with thorough research of the light power therapy, it should be clear about the photoactive activity constituent and the empoison mechanism especially, to formulate quality specification for the corresponding raw material.Pharmacognostic research of Bulgaria inquinans displayed that characteristic trait, microscopic trait, physichemical trait and molecular biology trait in this paper, The results showed that fruit bodies were snuff color and dotted with yellowish brown pits at the beginning, and then their upper side split into one or several pores, at last the surface of body turned black and crimpled. The asci are clubbed in shape, the ectal excipulum is made up of circular cells with hairs on the outside which contains chromophore-like substance, the medullary excipulum is consisted of interlaced mycelia imbedded in gelatine. The saffron yellow mycelia and septatus achromatous mycelia with growth clubbed conidium were gained by ascospore discharging method. The mycelia obtained by fruiting body tissue method were hyaline and filamentous, some mycelia composed of circular cell became bead-like, dark brown or yellowish. Comparing the DNA sequences from fruiting body and mycelia gain from liquid culture using strain of tissue culture, their similarity is 99.65%. After comparing the DNA sequences from fruiting body and AY789345 in GenBank, their similarity is 99.80%. The showing of organization separation obtains mushroom Spawn is stable and reliable, it is the pure mushroom spawn of Bulgaria inquinans.Fruiting body and fermental fruiting body of Bulgaria inquinans were extracted and separated in this article, 5 compounds and 1 component with photoactive activity were obtained in all, expertised 4 compounds, they are aremalonic acid, hexadecanoic acid, 4, 8-nonadecane dienoic acid and 2, 3-consecutive naphthyl-4-(1-hydroxy-normai-butyl)-5-valerolactone respectively. Posterior 3 compounds were separated from Bulgaria inquinans for the first time, 2, 3-consecutive naphthyl-4-(1-hydroxy-normal-butyl)-5- Valerolactone is a new compound. Photoactive activity component was predicated perylenequinones composition preliminarily.The Q component which obtained from the petroleum ether extract layer was used the GC-MS associated analysis, it mainly retrieved to 22 compounds. The G component which obtained from the chloroform extract layer was used the GC-MS associated analysis, it mainly retrieved to 7 compounds. They are mainly a kind of small polarity fatty group hydrocarbon and the fatty acid.The extract of acetone, extract of chloroform coming from extract of acetone and Y4 component and Y5 component which separated from the extract of chloroform displayed photoactive activity in photoactive activity experiments. Ear index displayed significant deviation to extract of acetone intragastric administration by 1000mg/kg for illumination series, ear index and skin index displayed extreme significant deviation to extract of acetone intragastric administration by 2000mg/kg for illumination series. Ear index and skin Index displayed extreme significant deviation to extract of chloroform intragastric administration by 260mg/kg and 520mg/kg for illumination series. Ear index displayed significant deviation to Y4 component intragastric administration by 200mg/kg for illumination series, ear index displayed extreme significant deviation to Y5 component intragastric administration by 200mg/kg and 100mg/kg for illumination series, skin Index displayed extreme significant deviation to Y4 component intragastric administration by 200mg/kg and Y5 component intragastric administration by 200mg/kg and 100mg/kg for illumination series. There is Certain dose-effect relationship in the photoactive activity and the dosage, its photoactive toxic attack also has certain dormiens period. Extract of acetone was used in rats through intragastric administration by 1g/kg for 5 days, illumination 2 hour every day, ears of rat started to fester, claws also became quite purple at fourth day, ears was disappeared by fester at sixth day. Photoactive activity of Bulgaria inquinans is also checked by experiment of rat. The locus that is sensitive to light is damaged acutely.更多还原