| Objective To research the mechanism of red blood cell damage(hemolytic activity)induced by the venom from the jellyfish Stomolophus meleagris,and to evaluate the effect of different concentrations of vitamins C on the hemolytic activity of the venom.Methods The nematocyst was separated from Stomolophus meleagris with the method of autolysis,and then we got the venom by breaking the nematocyst extracted by ultrasonic.The concentration of the venom protein was measured as a standard of bovine serum albumin.8 men and 8 women were selected as healthy volunteers,4ml of fresh blood was taken from each of them,red blood cells were washed for 2~3 times,and then20%(V/V)red blood cell suspension and 0.45%(V/V)human red blood cell suspension was dispensed.Different volume of the venom added into 1ml of 20% red blood cell suspension,and then added 0.9% Sodium Chloride Solution to 5ml,made the final concentration of the venom from Stomolophus meleagris of 0,2,4,8,16 μ g/ml.Malondialdehyde(MDA)level in hemolytic reaction system was detected after different concentration of the venom with the method of thiobarbituric(TBA).A certain volume of the venom was added into the 1ml of 0.45% red blood cell suspension,different volume of vitamin C was added,then added 0.9% Sodium Chloride Solution diluted to 5ml,the concentration of vitamin C was 0,0.5,1,1.5,2,2.5mmol/L,evaluated the effect of different concentrations of vitamins C on the hemolytic activity of the venom.Results Base on the data of this research,the concentration of the venom protein was395.45μg/ml.MDA level increased with dose the venom increased dependently in the hemolysis system of red blood cells by different concentrations of the venom.When the concentration of the venom is 16μg/ml,the concentration of MDA increased from baseline concentrations(0.79 ± 0.07 μmol/L)to(19.90 ± 1.22 μmol/L),there was statistical difference between the experimental group and the control group(p <0.05)by single factor analysis of variance and LSD-t test;there was statistical difference between experimental groups(p<0.05).A certain range of concentration of vitamin C could inhibit the activity of the venom,and the hemolytic activity decreased with the increase of the concentration of vitamin C.When the concentration of vitamin C was in the range of0~1mmol/L,the hemolytic activity of the venom decreased significantly from 75.79%(without adding vitamin C)to 31.95%;when the concentration of vitamin C was in the range of 1~2.5mmol/L,the hemolytic activity decreased from 31.95% to 26.66% with a slowly downward trend.There was statistical difference between the control group and the experimental group(p<0.05)by One way ANOVA and LSD-t test;the experimental group compared with each other,when the concentration of vitamin C is 1.5,2 and2.5mmol/L,there was no statistical difference(p>0.05);there was statistical difference between other groups(p<0.05).Conclusions The lipid peroxidation injury is one of the mechanisms of the hemolytic activity of the venom from Stomolophus meleagris.Vitamin C can inhibit the hemolytic activity of the venom to a certain extent,and it can be used as a candidate drug for the prevention and treatment of sting by the Stomolophus meleagris.There are some components which can produce lipid peroxidation damage to red blood cells,which provides experimental help for the purification of hemolysin protein.Significance To understand the mechanism of red cell injury caused by the venom from Stomolophus meleagris,and to provide experimental basis for the prevention and treatment of sting by the Stomolophus meleagris,so as to provide basic data for further study of bioactive substances in the venom. |
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