超低温冷冻对脊尾白虾精子几种酶活性的影响

Effects of cryopreservation on enzyme activity of Exopalaemon carinicauda spermatozoa

研究了超低温冷冻保存(-196℃)对脊尾白虾精子内琥珀酸脱氢酶(SDH)、乳酸脱氢酶(LDH)、Na+/K+-ATP酶、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽还原酶(GR)与顶体酶活性的影响,以期为提高脊尾白虾精子超低温冷冻效果提供理论依据。设置对照组(未添加抗冻剂)、3个实验组[分别添加DMSO(V/V)10.0%、12.5%、15.0%],于冷冻0、1、3、5、7、15 d取样测定各酶活性。结果表明,经冷冻后,除GR外,其他所测酶活性均出现显著下降(P<0.05),且以对照组酶活性下降幅度最大。GR活性在冷冻7 d内显著升高,且对照组明显高于实验组(P<0.05),在冷冻15 d时又出现下降。添加15.0%DMSO组所测酶活性均高于同期其它各组,表明15.0%DMSO对精子内酶的保护作用较好。冷冻15 d后15.0%DMSO组的SDH、LDH和Na+/K+-ATP酶活性由(28.500±1.453)U/mL、(1290.836±27.603)U/L和(2.605±0.232)μmol/(mg.h)分别降至(15.300±0.950)U/mL、(363.713±13.943)U/L和(0.542±0.186)μmol/(mg.h);SOD和CAT活性由(106.497±7.217)U/mL、(383.632±4.731)U/g分别降至(17.036±0.321)U/mL、(166.940±1.910)U/g;顶体酶活性从(3.521±0.010)μIU/106降至(1.212±0.043)μIU/106;而GR活性由(217.042±6.962)U/L上升至(302.787±24.558)U/L。从冷冻后各酶下降幅度来看,超低温冷冻对SOD活性的影响最大,其次是Na+/K+-ATP酶。

Exopalaemon carinicauda is one of the marine aquaculture species with high economic value in China.This paper studied the effects of cryopreservation（-196 ℃）on the enzyme activity（SDH,LDH,Na^＋K^＋-ATP,SOD,CAT,GR and Acrosin）in E.carinicauda spermatozoa,in order to provide a theoretical basis for cryopreservation effect on E.carinicauda spermatozoa,and lay a foundation for artificial propagation of E.carinicauda.We set up a control group（without cryoprotectant）,and three experimental groups（DMSO（V/V）10%,12.5%,15%）,detected the enzyme activities on day 0,1,3,5,7,15.The result showed that,after cryopreservation,in addition to GR,the other enzyme activities decreased significantly（P0.05）,with the biggest drop in the control group.GR activity significantly increased in 7 days,and the control group was significantly higher than the experimental group（P0.05）,declining again in 15 days.The group of 15% DMSO measured enzyme activities were higher than other groups,indicating that 15% DMSO had protective effect on enzyme of spermatozoa.15 days later,under the condition of 15% DMSO,the activities of SDH,LDH and Na^＋/K^＋-ATPase decreased from（28.500±1.453）U/mL,（1290.836±27.603）U/L and（2.605±0.232）μmol/（mg·h）to（15.300±0.950）U/mL,（363.713±13.943）U/L and（0.542±0.186）μmol/（mg·h）respectively;the activities of SOD and CAT decreased from（106.497±7.217）U/mL,（383.632±4.731）U/g to（17.036±0.321）U/mL and（166.940±1.910）U/g respectively;the activities of acrosin decreased from（3.521±0.010）μIU/106 to（1.212±0.043）μIU/106;but the activities of GR increased from（217.042±6.962）U/L to（302.787±24.558）U/L.From the decline range,cryopreservation had the biggest influence on SOD,followed by Na^＋/K^＋-ATPase.