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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 Snails after soaking in the extract of Melia azedarach leaves, to detect the activity of adenosine triphosphate enzyme, succinate dehydrogenase, lactate dehydrogenase and nitric oxide synthase. Methods 48h snails were collected from the extracts of Melia azedarach and frozen section, and the changes of four enzyme activities were detected by enzyme histochemical staining. The experimental group was treated with no drug. Results The gray value of Ca2 + -ATPase in the snail head and digestive gland of the experimental group was 0.197 ± 0.059 and 0.233 ± 0.037, respectively, which was significantly different from that of the control group (0.298 ± 0.041 and 0.413 ± 0.057) (t = 9.036 and 8.594, P <0.05). The gray value of SDH in experimental group was 0.152 ± 0.038, which was significantly different from that of control group (0.228 ± 0.041, t = 7.167, P <0.05) The gray value of NOS in foot and heart were 0.616 ± 0.064, 0.431 ± 0.043 and 0.716 ± 0.058, respectively, which were significantly different from those in control group (0.514 ± 0.049, 0.302 ± 0.042 and 0.503 ± 0.045, respectively 3.833, 6.125 and 8.608, P <0.05). Conclusion The levels of NOS and Ca2 + -ATPase and SDH in the snail treated with extract of Melia azedarach leaves decreased. Therefore, we speculated that the snail mechanism of Melia tomentosa could be caused by the increase of NOS, which increased the content of nitric oxide (NO) in the head of snail head, inhibited aerobic respiration in head and foot, decreased the activity of calcium pump, caused calcium overload and induced cell apoptosis, death.