Abstract: In order to explore the effects of insect hormones on the alkaline phosphatase(ALP) activity and its gene expression level in Bombyx mori,using the 3rd day of the fifth instar silkworm(Dazao) as test materials,the ALP gene expression level and enzyme activity were detected in the hemolymph and the midgut at different time after injecting 20E and JH.The results showed that the ALP expression level was significantly up-regulated at 6 h,12 h and 24 h after 20E or JH treatment in the hemolymph of Bombyx mori,but it was up-regulated only at 12 h and 24 h in the midgut. In the 20E treatment group,the ALP relative expression level was the highest at 6 h in the hemolymph,which was 2.3 times of the control group;but it was the highest at 12 h in the midgut,which was 2.8 times of the control group.After JH treatment,the ALP expression level in the hemolymph and midgut were the highest at 24 h,which were 3.7 times and 4.7 times of the control group respectively. In addition,the ALP enzyme activities were significantly activated in the hemolymph and midgut by 20E and JH.The hemolymph ALP enzyme activity was significantly higher than the control group at 6 h,12 h and 24 h after 20E treatment,and the highest activity at 12 h was 33.34 U/mg;While the activity of ALP in the midgut were significantly higher than the control group only at 12 h and 24 h,their activities were 37.68 U/mg and 40.95 U/mg respectively.Meanwhile,the ALP activity were also significantly higher in the hemolymph and midgut at 6 h,12 h and 24 h after JH treatment,among them,the ALP activity in the hemolymph had the highest value of 42.29 U/mg at 6 h,while the enzyme activity was rosed to the highest value of 46. 84 U/mg at 12 h in the midgut.The above results indicated that the ALP expression level was regulated by 20E and JH in Bombyx mori,and the effects of hormones on the gene expression level were consistent with the change rule of enzyme activity.

Key words: Insect hormone, Bombyx mori, Alkaline phosphatase, Enzymatic activity, Gene expression level, 20-hydroxyecdysone, Juvenile hormone

摘要： 为了探究昆虫激素对家蚕碱性磷酸酶（ALP）活性及其基因表达水平的影响，以五龄第3天家蚕（大造）幼虫为试验材料，注射外源蜕皮激素20E和保幼激素JH，分析检测家蚕血淋巴和中肠ALP基因及ALP酶活性的变化情况。结果显示，20E或JH处理后6 h、12 h和24 h，家蚕血淋巴ALP均明显上调表达，而中肠ALP仅在12 h和24 h上调表达。其中20E处理组，血淋巴ALP在6 h的相对表达量最高，为对照组的2.3倍；中肠ALP在12 h表达量最高，为对照组的2.8倍。JH处理组，血淋巴和中场ALP均是在24 h的相对表达量最高，分别是对照组的3.7倍和4.7倍。另外，20E和JH均能显著激活家蚕血淋巴和中肠的ALP酶活性。20E处理组，血淋巴ALP酶活性在20E处理6 h、12 h和24 h显著高于对照组；在12 h活性最高，为33.34 U/mg。中肠ALP酶活性是在12 h和24 h极显著高于对照组，其活性分别为37.68 U/mg和40.95 U/mg。JH处理组，血淋巴和中肠ALP酶活性在6 h，12 h和24 h显著高于对照组，其中血淋巴的ALP酶活性在6 h出现最高值(42.29 U/mg)，而中肠ALP酶活性在12 h升至最高值(46.84 U/mg)。上述结果表明，家蚕ALP基因受激素20E和JH的调控，且激素对ALP基因表达水平的影响与ALP酶活性变化规律一致。

关键词: 昆虫激素, 家蚕, 碱性磷酸酶, 酶活性, 基因表达水平, 蜕皮激素, 保幼激素