【作者】 邓星灿；

【导师】 王贤纯； 梁宋平；

【作者基本信息】 湖南师范大学 ， 生物化学与分子生物学， 2008， 硕士

【摘要】 急性胰腺炎是常见的急腹症之一,多见于青壮年。其发病仅次于急性阑尾炎、肠梗阻、急性胆囊炎胆石症。一般将急性胰腺炎分为急性水肿型(轻型)胰腺炎(占88-97%)和急性出血坏死型(重型)胰腺炎两种。急性出血坏死型约占3-12%,其病死率很高,达30-50%。本病误诊率高达60-90%。牛胰胰蛋白酶抑制剂(Bovine pancreatictrypsin inhibitor,BPTI)自60年代开始使用,发现它有对抗激肽和抗休克的作用,因而引起了临床医务工作者的高度重视。该药现已成为一种普遍治疗急性胰腺炎的药物,特别是对出血坏死型胰腺炎,能缓解病情,降低死亡率。从虎纹捕鸟蛛(Selenocosmia huwena Wang)粗毒中我们分离纯化到一种具有丝氨酸蛋白酶(如胰蛋白酶)抑制活性的多肽成分,命名为虎纹捕鸟蛛毒素Ⅺ(huwentoxin-Ⅺ,HWTX-Ⅺ)。这是首次报道从蜘蛛毒素中纯化到蛋白酶抑制剂。HWTX-Ⅺ属于BPTI/Kunitz型丝氨酸蛋白酶抑制剂家族,它的抑制活性比BPTI强。在本文的工作中,我们利用RACE的方法获得的HWTX-Ⅺ的cDNA序列,使用载体pVT102U实现了在酿酒酵母中的表达。HWTX-Ⅺ在酵母(Saccharomyces cerevisiae)中的表达产率为12.5mg/L。我们建立了一种腹腔分次大剂量注射L-Arg诱导的小鼠急性胰腺炎模型,该模型操作简单,重复性好。在该模型的基础上,我们探索HWTX-Ⅺ对急性胰腺炎的治疗作用。通过研究不同的剂量、不同的治疗时间对治疗效果的影响以及同抑肽酶治疗作用的比较得出,HWTX-Ⅺ是一种有效的治疗急性胰腺炎的药物。更多还原

【Abstract】 Acute pancreatitis is a common acute abdominal disease, mostly found among young and middle-aged people. The incidence of the disease is just lower than that of acute appendicitis, intestinal obstruction and acute cholecystitis. Acute pancreatitis can be classified into acute edematous pancreatitis ( light ) and acute nectotizing pancreatitis (heavy), and they account for 88-97% and 3-12% respectively. The mortality of acute nectotizing pancreatitis is very high, probably up to 30-50% and its misdiagnose rate is up to 60-90%. Bovine pancreatic trypsin inhibitor (BPTI) has been used since last century and received much attention from clinicians due to that it can work against shock and kinin. Now, it is a commonly drug used for the treatment of acute pancreatitis, especially the acute nectotizing pancreatitis because it can alleviate illness and reduce mortality. We discovered a new peptide from the venom of bird spider Selenocosmia huwena Wang, which, named huwentoxin-XI(HWTX-XI), has inhibitory activity against serine proteinases such as trypsin. Being the first proteinase inhibitor purified from spider venom, HWTX-XI is shown to belong to family of BPTI/Kunitz type of serine proteinase inhibitor, and has inhibitory ability stronger than BPTI. In the present work, we used the whole cDNA of HWTX-XI which was obtained by RACE method for expressing in Saccharomyces cerevisiae with PVT102V vector. The expression yield in Saccharomyces is up to 12. 5mg/L. We established a easily-operating and reproducible mouse model of acute pancreatitis induced by intraperitoneally injecting excessive dose of L-arginine. With this model, we investigated the cure effect of HWTX-XI on acute pancreatitis. By investigating the effects of different doses and cure time on cure effects as well as by comparing the cure efficiency of HWTX-XI and aprotinin, HWTX-XI was demonstrated as a drug that can effectively cure the acute pancreatitis.更多还原