加拿大一枝黄花石油醚萃取物对福寿螺肝脏的影响
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摘要
【目的】在前期研究的基础上深入探讨加拿大一枝黄花石油醚萃取物(petroleum ether extract of Solidago canadensis,PEEE)对福寿螺(Pomacea canaliculata)肝脏的影响,为进一步有效控制福寿螺对农业生产的危害、实现恶性外来入侵植物加拿大一枝黄花的资源化利用提供依据。【方法】以去氯自来水作为对照组,0.11、0.18和0.29 mg·mL-1的PEEE悬浮液作为试验组。每组随机放入大小、活力一致的成年福寿螺10只,用一层纱布和橡皮筋封住烧杯口,防止福寿螺逃逸。在室温(23—27℃)条件下,处理48 h,每组3次重复。48 h后,分别随机挑选去氯自来水、0.11、0.18和0.29 mg·mL-1浓度处理下的福寿螺,敲破螺壳,取出肝脏,使用扫描电镜和透射电镜观察肝脏组织的超微结构。经0.29 mg·mL-1浓度处理后的福寿螺肝脏组织冻干后用于代谢组学研究。【结果】扫描电镜观察结果表明,经不同浓度的PEEE浸泡处理福寿螺48 h后,福寿螺肝脏表面均受到损伤,其损伤程度与PEEE处理液的浓度呈正相关,高浓度的PEEE可致使肝脏萎缩失去弹性,皱褶凸起被侵蚀,体表破损,出现絮状物,质地松散。透射电镜观察结果表明,经不同浓度的PEEE浸泡处理福寿螺48 h后,福寿螺肝细胞损伤明显,高浓度的PEEE使福寿螺肝细胞粗面内质网及线粒体数量减少,粗面内质网出现肿胀、断裂和颗粒脱落等现象;线粒体表现为肿胀,形状不规则。代谢组学的试验结果表明,经PEEE处理后,福寿螺肝脏中60种代谢物含量发生变化,多个代谢通路受到影响,富集程度最高的两条通路分别为糖鞘脂代谢和棕榈酰辅酶A进入线粒体基质的通路。【结论】加拿大一枝黄花的石油醚萃取物对福寿螺肝脏细胞结构和生理功能有巨大的损伤破坏作用,因而具有较强的灭螺活性,有望开发成为新型的天然灭螺药物。
【Objective】 On the basis of previous studies, the effect of petroleum ether extract of Solidago canadensis(PEEE)on the liver of Pomacea canaliculata was discussed in depth to provide a basis for further effective control the harm of P.canaliculata to agricultural production and realizing the resource utilization of the malignant alien invasive plant S. canadensis.【Method】Chlorinated tap water was used as the control group, and PEEE suspensions of 0.11, 0.18 and 0.29 mg·mL~(-1) were used as the experimental groups. In each group, 10 adult P. canaliculata of the same size and vigor were randomly placed, and the beaker was sealed with a layer of gauze and rubber band to prevent the P. canaliculata from escaping. At room temperature(23-27℃), the treatment was carried out for 48 h with 3 repetitions in each group. After 48 h, the P. canaliculata treated with dechlorinated tap water, 0.11, 0.18 and 0.29 mg·mL~(-1) were randomly selected, the shells of P. canaliculata were broken and the liver was taken out, and the ultrastructure of liver tissue was observed by scanning electron microscopy and transmission electron microscopy. The liver tissue of P. canaliculata treated with 0.29 mg·mL~(-1) was lyophilized for metabonomics study. 【Result】The results of scanning electron microscopy showed that the liver surface of P. canaliculata was damaged after soaking for 48 h with different concentrations of PEEE, and the degree of damage was positively related to the concentration of PEEE treatment solution. High concentrations of PEEE could cause liver atrophy and loss of elasticity, wrinkle bulge erosion, body surface damage, flocs and loose texture. The results of transmission electron microscopy showed that the liver cells of P. canaliculata were damaged obviously after soaking for48 h with different concentrations of PEEE. The number of rough endoplasmic reticulum and mitochondria in liver cells of P.canaliculata decreased with high concentrations of PEEE, and the rough endoplasmic reticulum appeared swelling, breaking and degranulation. Mitochondria showed swelling and irregular shape. The results of metabonomic experiments showed that after PEEE treatment, the content of 60 metabolites in the liver of P. canaliculata changed, and several metabolic pathways were affected. The two pathways with the highest enrichment were glycosphingolipid metabolism and palmitoyl-CoA entering mitochondrial matrix pathway. 【Conclusion】PEEE of S. canadensis has great damage and destruction to the liver cells of P. canaliculata, so it has strong molluscicidal activity and is expected to be developed into a new natural molluscicidal drug with great potential development and application value.
【Objective】 On the basis of previous studies, the effect of petroleum ether extract of Solidago canadensis(PEEE)on the liver of Pomacea canaliculata was discussed in depth to provide a basis for further effective control the harm of P.canaliculata to agricultural production and realizing the resource utilization of the malignant alien invasive plant S. canadensis.【Method】Chlorinated tap water was used as the control group, and PEEE suspensions of 0.11, 0.18 and 0.29 mg·mL~(-1) were used as the experimental groups. In each group, 10 adult P. canaliculata of the same size and vigor were randomly placed, and the beaker was sealed with a layer of gauze and rubber band to prevent the P. canaliculata from escaping. At room temperature(23-27℃), the treatment was carried out for 48 h with 3 repetitions in each group. After 48 h, the P. canaliculata treated with dechlorinated tap water, 0.11, 0.18 and 0.29 mg·mL~(-1) were randomly selected, the shells of P. canaliculata were broken and the liver was taken out, and the ultrastructure of liver tissue was observed by scanning electron microscopy and transmission electron microscopy. The liver tissue of P. canaliculata treated with 0.29 mg·mL~(-1) was lyophilized for metabonomics study. 【Result】The results of scanning electron microscopy showed that the liver surface of P. canaliculata was damaged after soaking for 48 h with different concentrations of PEEE, and the degree of damage was positively related to the concentration of PEEE treatment solution. High concentrations of PEEE could cause liver atrophy and loss of elasticity, wrinkle bulge erosion, body surface damage, flocs and loose texture. The results of transmission electron microscopy showed that the liver cells of P. canaliculata were damaged obviously after soaking for48 h with different concentrations of PEEE. The number of rough endoplasmic reticulum and mitochondria in liver cells of P.canaliculata decreased with high concentrations of PEEE, and the rough endoplasmic reticulum appeared swelling, breaking and degranulation. Mitochondria showed swelling and irregular shape. The results of metabonomic experiments showed that after PEEE treatment, the content of 60 metabolites in the liver of P. canaliculata changed, and several metabolic pathways were affected. The two pathways with the highest enrichment were glycosphingolipid metabolism and palmitoyl-CoA entering mitochondrial matrix pathway. 【Conclusion】PEEE of S. canadensis has great damage and destruction to the liver cells of P. canaliculata, so it has strong molluscicidal activity and is expected to be developed into a new natural molluscicidal drug with great potential development and application value.
引文
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[9]吴淼生.江西农业外来有害生物及其防治.生物灾害科学,2013,36(4):434-437.WU M S.Prevention of agricultural exotic harmful organisms in Jiangxi Province.Biological Disaster Science,2013,36(4):434-437.(in Chinese)
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[12]ARCHIBALD R G.The use of the fruit of the tree Balanites?gyptiaca in the control of schistosomiasis in the Sudan.Transactions of the Royal Society of Tropical Medicine and Hygiene,1933,27(2):207-210.
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[14]刘席佑.加拿大一枝黄花抑螺活性及其挥发油的GC-MS分析研究[D].南昌:江西师范大学,2016.LI X Y.Studies on snails control activity and the GC-MS analysis of essential oil of Solidago canadensis[D].Nanchang:Jiangxi Normal University,2016.(in Chinese)
[15]WU J P,CHEN H C,HUANG D J.Histopathological and biochemical evidence of hepatopancreatic toxicity caused by cadmium and zinc in the white shrimp,Litopenaeus vannamei.Chemosphere,2008,73(7):1019-1026.
[16]PINHO G L,MOURA DA ROSA C,YUNES J S,LUQUET C M,BIANCHINI A,MONSERRAT J M.Toxic effects of microcystins in the hepatopancreas of the estuarine crab Chasmagnathus granulatus(Decapoda,Grapsidae).Comparative Biochemistry and Physiology Part C:Toxicology and Pharmacology,2003,135(4):459-468.
[17]DAI L P,WANG W X,DONG X J,HU R Y,NAN X Y.Molluscicidal activity of cardiac glycosides from Nerium indicum against Pomacea canaliculata and its implications for the mechanisms of toxicity.Environmental Toxicology and Pharmacology,2011,32(2):226-232.
[18]DE ASSUMP??O PEREIRA-DA-SILVA M,FERRI F A.1-Scanning electron microscopy//Nanocharacterization Techniques.William Andrew,2017:1-35.
[19]WANG Z L.Transmission electron microscopy of shape-controlled nanocrystals and their assemblies.The Journal of Physical Chemistry B,2000,104(6):1153-1175.
[20]NICHOLSON J K,LINDON J C.Systems biology:Metabonomics.Nature,2008,455(7216):1054-1056.
[21]NICHOLSON J K,CONNELLY J,LINDON J C,HOLMES E.Metabonomics:a platform for studying drug toxicity and gene function.Nature Reviews|Drug Discover,2002,1(2):153-161.
[22]SHEN X,WANG Z X,LIU L L,ZOU Z Z.Molluscicidal activity of Solidago canadensis L.extracts on the snail Pomacea canaliculata Lam.Pesticide Biochemistry and Physiology,2018,149:104-112.
[23]World Health Organization.Molluscicide screening and evaluation.Bulletin of the World Health Organization,1965,33(4):567-581.
[24]田硕,刘铜华,孙文,吴丽丽,秦灵灵,侯毅,董笑克,乐海波,洪明昭.夏枯草提取物对2型糖尿病ZDF大鼠肝糖原代谢的影响.中国实验方剂学杂志,2018,24(10):101-106.TIAN S,LIU T H,SUN W,WU L L,QIN L L,HOU Y,DONG X K,YUE H B,HONG M Z.Effect of Prunellae spica extracts on hepatic glycogen in ZDF rats.Chinese Journal of Experimental Traditional Medical Formulae,2018,24(10):101-106.(in Chinese)
[25]TELLA T,MASOLA B,MUKARATIRWA S.The effect of Psidium guajava aqueous leaf extract on liver glycogen enzymes,hormone sensitive lipase and serum lipid profile in diabetic rats.Biomedicine and Pharmacotherapy,2019,109:2441-2446.
[26]CHIANG J Y L.Liver physiology:Metabolism and detoxification//MCMANUS L M,MITCHELL R N.Pathobiology of Human Disease.Elsevier,2014:1770-1782.
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[28]ZHAO B L,FANG L,ZHANG J E,AN M,WANG R L,LUO M Z,LIANG K M.Effects of five exotic invasive plants extracts on the survival of the invasive snail Pomacea canaliculata(Lamarck).Allelopathy Journal,2015,36(1):123-132.
[29]闫小红,周兵,邹有,蒋平,许冬梅.16种植物提取物对福寿螺杀螺效果的研究.广东农业科学,2011(4):81-84.YAN X H,ZHOU B,ZOU Y,JIANG P,XU D M.Efficacy of the extracts from sixteen plants against Ampullaria gigas Spix.Guangdong Agricultural Science,2011(4):81-84.(in Chinese)
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[2]毕玉科,田旗,卢钟玲,李宏庆.舟山岛外来植物及其入侵性分析.福建林业科技,2015,42(1):151-159,172.BI Y K,TIAN Q,LU Z L,LI H Q.Analyses on invasiveness of exotic plants in Zhoushan Island,Zhejiang,China.Journal of Fujian Forestry Science and Technology,2015,42(1):151-159,172.(in Chinese)
[3]李小慧,胡隐昌,宋红梅,王培欣,汪学杰,牟希东,刘超,罗建仁.中国福寿螺的入侵现状及防治方法研究进展.中国农学通报,2009,25(14):229-232.LI X H,HU Y C,SONG H M,WANG P X,WANG X J,MU X D,LIU C,LUO J R.Invasion and monitoring methods of Pomacea canaliculata in China.Chinese Agricultural Science Bulletin,2009,25(14):229-232.(in Chinese)
[4]COWIE R H,BARKER G M.Apple snails(Ampullariidae)as agricultural pests:Their biology,impacts and management//Molluscs as Crop Pests.Wallingford,UK:CABI Publishing,2002:145-192.
[5]杨叶欣,胡隐昌,李小慧,汪学杰,牟希东,宋红梅,王培欣,刘超,罗建仁.福寿螺在中国的入侵历史、扩散规律和危害的调查分析.中国农学通报,2010,26(5):245-250.YANG Y X,HU Y C,LI X H,WANG X J,MU X D,SONG H M,WANG P X,LIU C,LUO J R.Historical invasion,expansion process and harm investigation of Pomacea canaliculate in China.Chinese Agricultural Science Bulletin,2010,26(5):245-250.(in Chinese)
[6]NAYLOR R.Invasions in agriculture:Assessing the cost of the golden apple snail in Asia.Ambio,1996,25(7):443-448.
[7]尉吉乾,莫建初,洪文英,徐文,许凤仙.广州管圆线虫中间宿主福寿螺的综合防治研究进展.中国媒介生物学及控制杂志,2010,21(5):512-514.WEI J Q,MO J C,HONG W Y,XU W,XU F X.Progress in integrated control of Pomacea canaliculata,intermediate host of Angiostrongylus cantonensis.Chinese Journal of Vector Biology and Control,2010,21(5):512-514.(in Chinese)
[8]钟锋.我国福寿螺防治技术研究进展.世界农药,2015,37(6):21-24.ZHONG F.Research progress on control technology of Pomacea canaliculata in China.World Pesticides,2015,37(6):21-24.(in Chinese)
[9]吴淼生.江西农业外来有害生物及其防治.生物灾害科学,2013,36(4):434-437.WU M S.Prevention of agricultural exotic harmful organisms in Jiangxi Province.Biological Disaster Science,2013,36(4):434-437.(in Chinese)
[10]毕研云,图立红,李枢强,刘月英.我国常用的灭螺方法.生物学通报,2006,41(11):17-18.BI Y Y,TU L H,LI S Q,LIU Y Y.The commonly used snail control methods in China.Bulletin of Biology,2006,41(11):17-18.(in Chinese)
[11]肖林飞,廖列文.新型灭螺剂的研究进展.广东化工,2012,39(4):26-27.XIAO L F,LIAO L W.Research progress of the new molluscicide.Guangdong Chemical Industry,2012,39(4):26-27.(in Chinese)
[12]ARCHIBALD R G.The use of the fruit of the tree Balanites?gyptiaca in the control of schistosomiasis in the Sudan.Transactions of the Royal Society of Tropical Medicine and Hygiene,1933,27(2):207-210.
[13]黄炳生,李明亚.植物源灭钉螺药的研究进展.中药材,2012,35(6):1010-1013.HUANG B S,LI M Y.Research progress of oncomelania killers from plants.Journal of Chinese Medicinal Materials,2012,35(6):1010-1013.(in Chinese)
[14]刘席佑.加拿大一枝黄花抑螺活性及其挥发油的GC-MS分析研究[D].南昌:江西师范大学,2016.LI X Y.Studies on snails control activity and the GC-MS analysis of essential oil of Solidago canadensis[D].Nanchang:Jiangxi Normal University,2016.(in Chinese)
[15]WU J P,CHEN H C,HUANG D J.Histopathological and biochemical evidence of hepatopancreatic toxicity caused by cadmium and zinc in the white shrimp,Litopenaeus vannamei.Chemosphere,2008,73(7):1019-1026.
[16]PINHO G L,MOURA DA ROSA C,YUNES J S,LUQUET C M,BIANCHINI A,MONSERRAT J M.Toxic effects of microcystins in the hepatopancreas of the estuarine crab Chasmagnathus granulatus(Decapoda,Grapsidae).Comparative Biochemistry and Physiology Part C:Toxicology and Pharmacology,2003,135(4):459-468.
[17]DAI L P,WANG W X,DONG X J,HU R Y,NAN X Y.Molluscicidal activity of cardiac glycosides from Nerium indicum against Pomacea canaliculata and its implications for the mechanisms of toxicity.Environmental Toxicology and Pharmacology,2011,32(2):226-232.
[18]DE ASSUMP??O PEREIRA-DA-SILVA M,FERRI F A.1-Scanning electron microscopy//Nanocharacterization Techniques.William Andrew,2017:1-35.
[19]WANG Z L.Transmission electron microscopy of shape-controlled nanocrystals and their assemblies.The Journal of Physical Chemistry B,2000,104(6):1153-1175.
[20]NICHOLSON J K,LINDON J C.Systems biology:Metabonomics.Nature,2008,455(7216):1054-1056.
[21]NICHOLSON J K,CONNELLY J,LINDON J C,HOLMES E.Metabonomics:a platform for studying drug toxicity and gene function.Nature Reviews|Drug Discover,2002,1(2):153-161.
[22]SHEN X,WANG Z X,LIU L L,ZOU Z Z.Molluscicidal activity of Solidago canadensis L.extracts on the snail Pomacea canaliculata Lam.Pesticide Biochemistry and Physiology,2018,149:104-112.
[23]World Health Organization.Molluscicide screening and evaluation.Bulletin of the World Health Organization,1965,33(4):567-581.
[24]田硕,刘铜华,孙文,吴丽丽,秦灵灵,侯毅,董笑克,乐海波,洪明昭.夏枯草提取物对2型糖尿病ZDF大鼠肝糖原代谢的影响.中国实验方剂学杂志,2018,24(10):101-106.TIAN S,LIU T H,SUN W,WU L L,QIN L L,HOU Y,DONG X K,YUE H B,HONG M Z.Effect of Prunellae spica extracts on hepatic glycogen in ZDF rats.Chinese Journal of Experimental Traditional Medical Formulae,2018,24(10):101-106.(in Chinese)
[25]TELLA T,MASOLA B,MUKARATIRWA S.The effect of Psidium guajava aqueous leaf extract on liver glycogen enzymes,hormone sensitive lipase and serum lipid profile in diabetic rats.Biomedicine and Pharmacotherapy,2019,109:2441-2446.
[26]CHIANG J Y L.Liver physiology:Metabolism and detoxification//MCMANUS L M,MITCHELL R N.Pathobiology of Human Disease.Elsevier,2014:1770-1782.
[27]宋春秀,章家恩,罗明珠,赵本良,何铭谦.薇甘菊浸提液对福寿螺主要器官组织损伤的扫描电镜观察.生态环境学报,2010,19(8):1876-1880.SONG C X,ZHANG J E,LUO M Z,ZHAO B L,HE M Q.Damage of extracts from Mikania micrantha on tissues of golden apple snails(Pomacea canaliculata).Ecology and Environmental Sciences,2010,19(8):1876-1880.(in Chinese)
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