放射线对体外培养棘球蚴及其Gadd45α mRNA表达的影响

Effects of radiation on echinococcus in vitro culture and its mRNA expression of Gadd45α

目的观察放射线对体外培养棘球蚴的杀伤作用及对其生长阻滞和DNA损伤45α(growth arrest and DNA damage 45 alpha,Gadd45α)mRNA表达的影响。方法体外培养自然感染的羊肝脏中的棘球蚴,分为7组,分别以0(对照)、20、40、60、80、100、120 Gy的6兆电子线(6 MeV)进行1次放射。放射线照射后培养1、3、5、7 d时,光镜下观察棘球蚴的生长情况;采用反转录-聚合酶链反应(RT-PCR)法测定放射线照射后培养7 d时,对照,20、40、60 Gy组棘球蚴Gadd45αmRNA表达水平。结果各剂量组放射线照射后1、3、5、7 d时,光镜下棘球蚴结构崩解,勾突脱落,且棘球蚴死亡率与放射线照射剂量呈正相关(r=0.81,P<0.05)。放射线照射后培养7 d时,与对照组(100.00±0.00)比较,20、40、60 Gy组棘球蚴Gadd45αmRNA表达水平明显升高(279.74±80.08、759.38±160.98、1666.68±316.36,P均<0.01),且与放射线剂量呈正相关(r=0.93,P<0.01)。结论放射线照射对体外培养的棘球蚴有一定的杀伤作用,且与放射线的剂量存在一定的量效关系;Gadd45α基因可能参与放射线所致体外杀伤棘球蚴的分子机制。

Objective To investigate the killing effect of radiation on echinococcus in vitro culture and its effect on the mRNA expression of growth arrest and DNA damage 45 alpha(Gadd45α)gene.Methods Echinococcus from naturally infected sheep liver was cultured in vitro and divided into 7 groups.The echinococcus was irradiated with 6 MeV at doses of 0(control),20,40,60,80,100,and 120 Gy,respectively.The growth of echinococcus was observed under light microscope at 1,3,5 and 7 d after the radiation.The expression of Gadd45αmRNA in control,20,40 and 60 Gy groups of echinococcus was detected by RT-PCR technique at 7 d after the radiation.Results The disintegration and exfoliation of echinococcus were observed under light microscope at 1,3,5 and 7 d after the radiation,and the death rate of echinococcus was positively correlated with the radiation dosages(r=0.81,P<0.05).After the radiation at 7 d,compared with the control group(100.00±0.00),the mRNA expression levels of Gadd45αin echinococcus of 20,40,and 60 Gy groups were significantly increased(279.74±80.08,759.38±160.98,1666.68±316.36,P<0.01),and it was positively correlated with the radiation dosages(r=0.93,P<0.01).Conclusion Radiation has a certain killing effect on echinococcus cultured in vitro,and there is a certain dose-effect relationship with the radiation dosages,and Gadd45αgene may be involved in the molecular mechanism of radiation-induced killing of echinococcus in vitro.