Zhang Hui, Wang Qianqian, Xiao Liang, Zhang Liming
Department of Marine Biotechnology, Faculty of Naval Medicine, Second Military Medical University, Shanghai, China.
PeerJ. 2017 May 9;5:e3338. doi: 10.7717/peerj.3338. eCollection 2017.
Cations have generally been reported to prevent jellyfish venom-induced hemolysis through multiple mechanisms by spectrophotometry. Little attention has been paid to the potential interaction between cations and hemoglobin, potentially influencing the antagonistic effect of cations. Here, we explored the effects of five reported cations, La, Mn, Zn, Cu and Fe, on a hemolytic test system and the absorbance of hemoglobin, which was further used to measure their effects on the hemolysis of tentacle extract (TE) from the jellyfish . All the cations displayed significant dose-dependent inhibitory effects on TE-induced hemolysis with various dissociation equilibrium constant () values as follows: La 1.5 mM, Mn 93.2 mM, Zn 38.6 mM, Cu 71.9 μM and Fe 32.8 mM. The transparent non-selective pore blocker La did not affect the absorbance of hemoglobin, while Mn reduced it slightly. Other cations, including Zn, Cu and Fe, greatly decreased the absorbance with values of 35.9, 77.5 and 17.6 mM, respectively. After correction, the inhibitory values were 1.4 mM, 45.8 mM, 128.5 μM and 53.1 mM for La, Zn, Cu and Fe, respectively. Mn did not inhibit TE-induced hemolysis. Moreover, the inhibitory extent at the maximal given dose of all cations except La was also diminished. These corrected results from spectrophotometry were further confirmed by direct erythrocyte counting under microscopy. Our results indicate that the cations, except for La, can interfere with the absorbance of hemoglobin, which should be corrected when their inhibitory effects on hemolysis by jellyfish venoms are examined. The variation in the inhibitory effects of cations suggests that the hemolysis by jellyfish venom is mainly attributed to the formation of non-selective cation pore complexes over other potential mechanisms, such as phospholipases A2 (PLA2), polypeptides, protease and oxidation. Blocking the pore-forming complexes may be a primary strategy to improve the in vivo damage and mortality from jellyfish stings due to hemolytic toxicity.
一般报道称，阳离子可通过分光光度法的多种机制预防水母毒液诱导的溶血。阳离子与血红蛋白之间的潜在相互作用很少受到关注，这可能会影响阳离子的拮抗作用。在此，我们探究了五种已报道的阳离子（镧、锰、锌、铜和铁）对溶血测试系统和血红蛋白吸光度的影响，并进一步用于测量它们对水母触手提取物（TE）溶血的影响。所有阳离子对TE诱导的溶血均表现出显著的剂量依赖性抑制作用，其解离平衡常数（）值如下：镧1.5 mM、锰93.2 mM、锌38.6 mM、铜71.9 μM和铁32.8 mM。透明的非选择性孔道阻滞剂镧不影响血红蛋白的吸光度，而锰使其略有降低。其他阳离子，包括锌、铜和铁，分别使吸光度大幅降低，其值分别为35.9、77.5和17.6 mM。校正后，镧、锌、铜和铁的抑制值分别为1.4 mM、45.8 mM、128.5 μM和53.1 mM。锰不抑制TE诱导的溶血。此外，除镧外，所有阳离子在最大给定剂量下的抑制程度也有所降低。分光光度法的这些校正结果通过显微镜下直接红细胞计数得到进一步证实。我们的结果表明，除镧外，阳离子会干扰血红蛋白的吸光度，在检测它们对水母毒液溶血的抑制作用时应进行校正。阳离子抑制作用的变化表明，水母毒液引起的溶血主要归因于非选择性阳离子孔道复合物的形成，而非其他潜在机制，如磷脂酶A2（PLA2）、多肽、蛋白酶和氧化作用。阻断成孔复合物可能是改善水母蜇伤因溶血毒性导致的体内损伤和死亡率的主要策略。
