本文選題：湖北釘螺 + WPN�。� 參考：《武漢大學》2016年博士論文

【摘要】：日本血吸蟲病是廣泛流行于我國的人獸共患寄生蟲病。湖北釘螺(Oncomelania hupensis)是日本血吸蟲(Schistosoma japonicum)唯一的的中間宿主,在血吸蟲病的傳播中起著十分重要的作用�？刂漆斅菔茄x病綜合防治策略中的重要措施,其中化學藥物滅螺具有高效性和使用方法簡單的特點,仍是目前最常用的滅螺手段,在控制釘螺的密度方面發(fā)揮重要作用。氯硝柳胺(WPN)是目前常用的,并被世界衛(wèi)生組織推薦用于現(xiàn)場滅螺的唯一藥物,但其滅螺機理仍未完全闡明。新型滅螺藥氯代水楊胺(lv dai shui yang an, LDS)是WPN的衍生物,經(jīng)實驗室和現(xiàn)場實驗證實其與WPN具有相當?shù)臏缏菪Ч?且具有成本低、對水生生物毒性小等優(yōu)點。本文在組織學、酶學以及轉(zhuǎn)錄組學水平比較研究釘螺在LDS作用后與WPN、水作用后的差異,以揭示LDS的殺螺機理與WPN的異同。通過浸泡法使用WPN、LDS和水對釘螺浸殺3h、6h、12h、24h后,在確定其殺螺效果的基礎上,利用HE染色和常規(guī)透射電鏡觀察其組織學變化,結(jié)果顯示,兩種藥物的作用均造成肌肉細胞內(nèi)肌動蛋白纖維的紊亂,細胞核、線粒體、內(nèi)質(zhì)網(wǎng)等細胞器的損傷,其中LDS處理組中可以觀察到肌肉細胞中更嚴重的細胞核損傷以及肝臟細胞更嚴重的空泡化及內(nèi)質(zhì)網(wǎng)斷裂現(xiàn)象。這些損傷表明在藥物處理后釘螺的運動能力以及肝臟和能量代謝均受到抑制。通過酶組織化學染色以及軟體組織勻漿測酶活的方法對乳酸脫氫酶(lactate dehydrogenase, LDH)等13種酶在兩種藥物處理后的活性變化進行檢測。在兩種滅螺藥的作用下,糖類代謝相關的酶類活性除了肌肉中的LDH均受到抑制,應激反應相關的酶類活性相對于對照組均有先上升后下降的趨勢。糖類代謝相關酶類活性的變化表明在兩種藥物處理下釘螺的糖類代謝模式出現(xiàn)改變,引起能量供應不足以及乳酸的堆積；應激反應相關的酶類活性變化則提示在藥物處理的早期釘螺具有較強的應激反應以及在藥物處理的后期肝臟以及神經(jīng)系統(tǒng)的嚴重損傷。大體來說WPN和LDS的作用效果是類似的,均造成了釘螺的組織結(jié)構(gòu)、神經(jīng)信號傳導、能量代謝、應激水平以及肝臟功能等方面的損傷或變化,但LDS處理后的釘螺肝臟表現(xiàn)出更嚴重的損傷,應激水平和有氧呼吸相關的酶活性也受到更強的抑制。在上述研究的基礎上,進一步利用轉(zhuǎn)錄組測序篩選出了WPN/LDS處理后差異表達的254個unigenes。GO和KEGG分析表明這些差異表達unigenes與細胞結(jié)構(gòu)病變、神經(jīng)信號傳導以及能量代謝的抑制有關,這與酶活性變化的結(jié)果是相符的。然而,考慮到轉(zhuǎn)錄后調(diào)控和蛋白水平調(diào)控對酶活性的影響,對潛在的酶相關基因及調(diào)控模式的完整鑒定和注釋在對LDS的滅螺機理的探索過程中非常必要的。這個轉(zhuǎn)錄組數(shù)據(jù)能夠為湖北釘螺在分子水平上的深入研究提供幫助,促進湖北釘螺基因組的研究,其中與滅螺機制相關的數(shù)據(jù)可以給將來釘螺的控制提供巨大的幫助。
[Abstract]:Schistosomiasis (Oncomelania hupensis), the only intermediate host of Schistosoma japonicum (Schistosoma japonicum), plays a very important role in the transmission of schistosomiasis. Control of Oncomelania is an important measure in the comprehensive prevention and control strategy of blood fluke disease. It is still the most commonly used means of snail control to control the density of Oncomelania Snail, which is the most commonly used method to control the density of Oncomelania snails. WPN is currently used commonly, and is recommended by WHO for the only drug used in the field snail. However, the mechanism of the snail control is still not fully elucidated. LV Dai Shui Yang an (LDS) is a derivative of WPN. Through laboratory and field experiments, it has been proved that it has considerable snail effect with WPN, and has the advantages of low cost and low toxicity to aquatic organisms. In this paper, the differences of Oncomelania snails after LDS action with WPN and water after the action of LDS were compared in the histology, enzymology and transcriptional level. The mechanism of snail killing of LDS is different from that of WPN. After immersing the snail with WPN, LDS and water to kill 3h, 6h, 12h, 24h by immersion method, on the basis of determining its snail killing effect, the histological changes are observed by HE staining and conventional transmission electron microscopy. The results show that the effects of the two kinds of drugs all cause the disorder of actin fiber in the muscle cells, the nucleus, The damage of mitochondria, endoplasmic reticulum and other organelles, in which the more serious nuclear damage in the muscle cells and the more severe vacuolization and endoplasmic reticulum fracture in the liver cells were observed in the LDS treatment group. These damage showed that the movement ability of Oncomelania and the liver and energy metabolites were suppressed after the treatment. Chemical staining and soft tissue homogenate measure enzyme activity to detect the activity changes of 13 enzymes, such as lactate dehydrogenase (lactate dehydrogenase, LDH), and other drugs treated by two drugs. Under the action of two kinds of snail, the enzyme activity related to carbohydrate metabolism is inhibited in addition to the LDH in the muscle, and the activity related enzyme activity The changes in carbohydrate metabolism related enzymes showed that the carbohydrate metabolism patterns of Oncomelania snails were changed under the treatment of two kinds of drugs, which resulted in the lack of energy supply and the accumulation of lactic acid. The changes in the activity of enzymes related to stress reaction suggested that the early Oncomelania snails were stronger in the drug treatment. Stress response and severe damage to the liver and nervous system at later stage of drug treatment. In general, the effects of WPN and LDS are similar, causing damage or change in the structure, nerve signal, energy metabolism, stress level and liver function of Oncomelania oncomelania, but the liver of Oncomelania snails treated by LDS is shown. On the basis of the above study, 254 unigenes.GO and KEGG analyses of differential expression after WPN/LDS treatment showed that these differences expressed unigenes and cell structural changes, neural signal transduction and the more severe injury. The inhibition of energy metabolism is related to the results of changes in enzyme activity. However, considering the effect of post transcriptional regulation and protein level regulation on enzyme activity, the complete identification and annotation of potential enzyme related genes and regulation patterns are necessary in the exploration of the mechanism of LDS. The deep research on the molecular level of Oncomelania Snail in Hubei provides help to promote the study of the genome of Oncomelania Snail in Hubei. The data related to the mechanism of snail mechanism can provide great help for the control of Oncomelania snails in the future.
【學位授予單位】：武漢大學
【學位級別】：博士
【學位授予年份】：2016
【分類號】：R184.38

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