發(fā)布時(shí)間：2018-03-16 06:21

  本文選題：新斯的明　切入點(diǎn)：銀環(huán)蛇毒素　出處：《廣州醫(yī)科大學(xué)》2013年碩士論文　論文類型：學(xué)位論文

【摘要】：背景和目的 我國每年蛇傷病例約30萬例，病死率3-5%。特別是華南地區(qū)，毒蛇傷是常見急癥之一，其中銀環(huán)蛇咬傷占各類毒蛇咬傷的8.12%[1，2]。銀環(huán)蛇毒素以神經(jīng)毒素為主[3,4,5]，干毒包含多種成分，如α-銀環(huán)蛇毒素、β-銀環(huán)蛇毒素、κ-銀環(huán)蛇毒素、P-銀環(huán)蛇毒素[6,7]等，毒性強(qiáng)，中毒者常因神經(jīng)肌肉傳導(dǎo)阻滯表現(xiàn)為的骨骼肌無力，臨床骨骼肌無力癥狀出現(xiàn)的順序?yàn)橄纫娧鄄�下垂、后可有吞咽困難、流涎，繼而四肢肌力減弱甚至癱瘓，嚴(yán)重者可因呼吸肌麻痹造成窒息死亡,癥狀恢復(fù)順序與出現(xiàn)順序相反。 目前國際公認(rèn)的治療蛇咬傷的方法是傷口局部處理及盡早使用抗蛇毒血清，嚴(yán)重者影響到呼吸功能時(shí)需要機(jī)械輔助通氣、血液灌流及恢復(fù)肌力的藥物等治療。鑒于蛇咬傷常發(fā)生在山區(qū)、郊外等地，，地方醫(yī)院不能常規(guī)配備抗蛇毒血清，使得抗蛇毒血清無法得到普遍推廣和應(yīng)用。故尋求更多的治療方法即能安全、廣泛的使用，又能為蛇咬傷病人爭取更多的救治時(shí)間是十分必要的。在以往的報(bào)道中，新斯的明拮抗銀環(huán)蛇成分毒素（例如：α-銀環(huán)蛇毒素、 β-銀環(huán)蛇毒素等）或其他蛇毒神經(jīng)毒素所致骨骼肌無力的影響尚存爭議[8]。又有相關(guān)研究表明，新斯的明在拮抗其他因神經(jīng)肌肉傳導(dǎo)阻滯而出現(xiàn)骨骼肌無力的疾病中療效確切，例如非去極化肌松劑所致的骨骼肌無力[9,10，11]。曾有試驗(yàn)將多種蛇毒神經(jīng)毒素與非去極化肌松劑（如筒箭毒堿）進(jìn)行對(duì)比[8]，多項(xiàng)結(jié)果顯示其致病機(jī)制是及其相似的。而目前有關(guān)新斯的明拮抗銀環(huán)蛇干毒所致骨骼肌無力的影響尚鮮有實(shí)驗(yàn)論證。偶有臨床報(bào)道新斯的明在治療神經(jīng)毒素為主的蛇咬傷疾病時(shí)可減少呼吸機(jī)的應(yīng)用[12]。 本項(xiàng)研究旨在：通過檢測不同濃度新斯的明對(duì)經(jīng)銀環(huán)蛇毒素處理后的大鼠膈肌肌條張力的改變，明確新斯的明是否可改善骨骼肌肌力。 方法 將健康SPF級(jí)雌性SD大鼠20只，隨機(jī)分為四組（n=5）:正常對(duì)照組（A組）、1μM新斯的明（B組）、3μM新斯的明（C組）、10μM新斯的明（D組）。A組給予生理鹽水作為正常對(duì)照組。實(shí)驗(yàn)大鼠均經(jīng)戊巴比妥鈉腹腔麻醉，取出膈肌并去除周圍的殘留血液、脂肪組織等后，將膈肌沿肌纖維方向分離制成實(shí)驗(yàn)用肌條（約1cmX1.5cm）。肌條一端連于高精度張力換能器，通過放大器由生理記錄儀記錄張力大小。另一端固定于固定桿上并使整個(gè)肌條浸于注有苛氏液的麥?zhǔn)喜壑�，槽中持續(xù)通入混合氣（95%O2，5%CO2），溫度維持37℃。固定后持續(xù)調(diào)節(jié)膈肌肌條前負(fù)荷為1g，平衡45-60分鐘，每15分鐘更換苛氏液一次[8]。平衡后用10-4M濃度的乙酰膽堿刺激兩次，待再次達(dá)到平衡后，最終選擇肌條張力維持在＜（1±10%）g[13]范圍內(nèi)的肌條作為實(shí)驗(yàn)用肌條。繼續(xù)給予銀環(huán)蛇干毒毒素（濃度：5μg/ml），5分鐘后沖洗并給予不同濃度的新斯的明（1μM、3μM、10μM）或生理鹽水（對(duì)照組)。記錄不同時(shí)間段（5min、10min、15min、30min、60min）膈肌肌條張力(g)大小。 結(jié)果 不同時(shí)間段A組、B組、C組、D組的肌條張力如下,5min時(shí)分別為:（0.846±0.019）g、（0.853±0.027）g、（0.853±0.024）g、（0.858±0.013）g；10min時(shí)分別為：（0.844±0.023）g、（0.860±0.033）g、（0.863±0.030）g、（0.912±0.026）g；15min時(shí)分別為：（0.846±0.033）g、（0.870±0.033）g、（0.873±0.024）g、（0.920±0.025）g；30min時(shí)分別為：（0.848±0.019）g、（0.870±0.029）g、（0.888±0.036）g、（0.932±0.013）g；60min時(shí)分別為：（0.860±0.024）g、（0.883±0.031）g、（0.908±0.032）g、（0.954±0.015）g。 （1）與對(duì)照組（A組）比較：B組與A組在各時(shí)間段檢測結(jié)果均無統(tǒng)計(jì)學(xué)差異（P0.05）;C組與A組在60min時(shí)檢測結(jié)果存在統(tǒng)計(jì)學(xué)差異（P 0.05）。D組與A組在10min、15min、30min、60min時(shí)檢測結(jié)果存在統(tǒng)計(jì)學(xué)差異（P≤0.01）。 （2）與B組比較：C組與B組在各時(shí)間段檢測結(jié)果均無統(tǒng)計(jì)學(xué)差異（P0.05）;D組與B組在10min、15min、30min、60min時(shí)檢測結(jié)果存在統(tǒng)計(jì)學(xué)差異。（10min、15min時(shí)，P 0.05；30min、60min時(shí)，P≤0.01）。 （3）與C組比較：D組與C組在60min時(shí)檢測結(jié)果存在統(tǒng)計(jì)學(xué)差異（P 0.05）。 （4）實(shí)驗(yàn)各組肌條的張力在不同時(shí)間段均有不同程度的恢復(fù)，但在60分鐘時(shí)均未完全恢復(fù)至最初水平。 結(jié)論 新斯的明拮抗銀環(huán)蛇毒素所致離體大鼠膈肌肌條無力的作用明確，且效應(yīng)與劑量有相關(guān)性。
[Abstract]:Background and purpose
China's annual snakebite cases about 300 thousand cases, the mortality rate of 3-5%. especially in the Southern China area, the snake injury is one of the most common emergency cases, which accounted for 8.12%[1,2]. of all kinds of snake bites k-bungarotoxin Snakebite with neurotoxin based [3,4,5], contains a variety of ingredients such as dry poison, alpha bungarotoxin, beta bungarotoxin, kappa bungarotoxin, P- bungarotoxin [6,7], strong toxicity, skeletal muscle weakness poisoning often caused by neuromuscular block as clinical symptoms, skeletal muscle weakness in the order of the seer ptosis, can have difficulty swallowing saliva flow, and limb muscle weakness and even paralysis, severe cases can cause suffocation due to respiratory muscle paralysis symptoms, restore order with the emergence of the opposite order.
At present, the internationally recognized methods to cure snake bite is a local wound treatment and early use of antivenom, seriously affect the respiratory function need mechanical ventilation, blood perfusion and recovery of muscle strength in drug therapy. In view of snake bites often occur in the mountains, on the outskirts of other places, the local hospital is not equipped with conventional anti venom serum. The antivenom could not be widely promoted and applied. Therefore, seeking more treatment is safe, and can be widely used for treatment of snakebite patients more time is necessary. In the previous report, the composition of neostigmine antagonism YinHuanShe toxins (e.g. alpha bungarotoxin. Bungarotoxin etc.) [8]. effect is controversial or other venom neurotoxin induced bone gravis and related research shows that neostigmine antagonism in the neuromuscular block out The effect of skeletal muscle weakness in diseases such as the exact, non depolarizing muscle relaxant caused by skeletal muscle weakness [9,10,11]. had multiple test venom neurotoxin and non depolarizing muscle relaxant (such as tubocurarine) compared to [8], a number of results showed that the pathogenic mechanism is similar. And the influence of the antagonistic related YinHuanShe Ming neostigmine dry skeletal muscle weakness caused by poison is rarely withexperiment. Occasionally reports of clinical application of [12]. based neurotoxin in the treatment of neostigmine snakebite disease can reduce ventilator
This study aims to: to detect different concentrations of neostigmine on the tension of k-bungarotoxin treated rat diaphragm muscle through the change of clear, neostigmine can improve skeletal muscle strength.
Method
灝嗗仴搴稴PF綰ч泴鎬D澶ч紶20鍙

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