苦楝叶对钉螺酶组织化学的影响  

The effect of Melia azedarach on enzyme histochemistry in Oncomelania hupensis

作　　者：顾文彪[1] 张仪[1] 夏尚光[2] 吕山[1] 朱丹[1] 吴缨[1] 刘和香[1] 王龙杰[1] 

机构地区：[1]中国疾病预防控制中心寄生虫病预防控制所、世界卫生组织疟疾、血吸虫病和丝虫病合作中心、卫生部寄生虫病重点实验室,上海200025 [2]安徽省林业科学研究院,安徽合肥230031

出　　处：《中国病原生物学杂志》2014年第1期16-19,共4页Journal of Pathogen Biology

基　　金：国家“十二五”国家科技计划项目(No.2011BAD38B070101);国家科技重大专项(No.2012ZX10004220,2008ZX10004-011)

摘　　要：目的钉螺经苦楝叶浸提液浸泡后,检测其体内三磷酸腺苷酶、琥珀酸脱氢酶、乳酸脱氢酶和一氧化氮合酶的活性,探讨苦楝叶杀螺机制。方法经苦楝叶浸提液处理48h的钉螺,冰冻切片,使用酶组织化学染色检测4种酶活性的变化,实验设未经药物处理对照组。结果实验组钉螺头足部和消化腺的Ca2+-ATPase灰度值分别为0.197±0.059和0.233±0.037,与对照组0.298±0.041和0.413±0.057比较差异有统计学意义(t=9.036和8.594,P<0.05);实验组钉螺头足部肌纤维的SDH灰度值为0.152±0.038,与对照组0.228±0.041比较差异有统计学意义(t=7.167,P<0.05);实验组钉螺神经中枢、足部和心脏的NOS灰度值分别为0.616±0.064、0.431±0.043和0.716±0.058,与对照组分别为0.514±0.049、0.302±0.042和0.503±0.045比较差异均有统计学意义(t分别为3.833、6.125和8.608,P<0.05)。结论经苦楝叶浸提液处理的钉螺NOS水平升高Ca2+-ATPase和SDH水平下降。因此推测钉螺苦楝叶杀螺机制可能是NOS升高增加了钉螺头足部NO的含量,从而抑制头足部有氧呼吸,降低钙泵的活性,引起钙超载,诱导细胞凋亡,最终导致钉螺死亡。Objective To explore the molluscicidal mechanism of Melia azedarach, the activity of Ca2＋-ATPase, succi- nate dehydrogenase （SDH）, lactate dehydrogenase （LDH）, and nitric oxide synthase （NOS） were detected in Oncomela nia hupensis that had been immersed in a leaf extract of Melia azedarach. Methods Snails in the experimental group were immersed in the leaf extract of M. azedarach for 48 hours and soft tissues were separated for preparation of frozen sections （6-μm slices）. Levels of Ca2＋-ATPase, SDH, LDH, and NOS activity were detected using an enzyme histo- chemical technique. Untreated snails served as controls. Results Ca2 ＋ -ATPase staining revealed that mean gray density values for the head foot and digestive gland differed significantly （P〈0.05） between the experimental group （0. 197± 0.059, 0. 233±0. 037） and control group （0.298±0.041, 0.413±0.057）. SDH staining suggested that the mean gray density value for muscle differed significantly （P〈0.05） between the experimental group （0. 152±0. 038） and control group （0. 228±0. 041）. NOS staining revealed that the mean gray density values for the ganglion, foot, and heart dif fered significantly （P〈0. 05） between the experimental group （0. 616±0. 064, 0. 431±0. 043, and 0. 716±0. 058） and control group （0.514±0.049, 0.302±0.042, and 0. 503±0. 045）. Conclusion The enzymatic activity of both Ca2＋- ATPase and SDH declined, and SDH activity rose in snails after treatment with a leaf extract of M. azedarach. The mol- luscicidal mechanism of M. azedarach is presumably by increasing the content of NO in the head-foot, thus inhibiting aer obic respiration by reducing calcium pump activity. This results in a calcium overload that induces apoptosis and eventual ly leads to death.

关 键 词：苦楝 钉螺 酶组织化学 三磷酸腺苷酶 琥珀酸脱氢酶 乳酸脱氢酶 一氧化氮合酶 

分 类 号：R184.38[医药卫生—流行病学]