2，4-二氯苯酚和壬基基酚对家蚕的环境激素作用研究
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摘要
目的 为了探讨环境激素对鳞翅目昆虫的影响,研究了环境激素2,4—二氯苯酚(2,4-DCP)和壬基酚(NP)对家蚕机体、BmN和Sf9培养细胞及家蚕培养造血器官的作用。
方法 通过添加含有环境激素2,4-DCP和NP的人工饲料来饲养家蚕,系统调查家蚕的生长速度、生长量、变态、生命力和生殖情况。TC-100培养基中添加2,4-DCP或NP,体外培养BmN、Sf9细胞和家蚕造血器官,进行培养BmN细胞的DNA损伤和造血器官功能调查,用MTT法调查细胞增殖。
结果 家蚕幼虫食下含0.8 mmol·L~(-1)以上浓度的2,4-DCP或0.500 mmol·L~(-1)以上浓度的NP饲料,生长速度、生长量、发育整齐度、生命力和生殖能力都明显下降;生殖能力所受影响应性别而不同;家蚕发育后期和蜕皮、化蛹、羽化等变态时期影响更加明显,在蚕体内可能有累积作用。体内和被污染后环境中的浓度水平2,4-DCP和NP对家蚕具有环境激素作用。
2,4-DCP和NP对BmN或Sf9细胞增殖的影响有一定差异,不同浓度的影响也不同;2种细胞对2,4-DCP的IC_(50)约为0.14 mmol·L~(-1),对NP约为0.05 mmol·L~(-1)。0.128mmol·L~(-1)以上浓度的NP,能诱导家蚕BmN细胞DNA链发生断裂,导致DNA损伤,而且具有较明显的浓度效应。NP的浓度超过0.032 mmol·L~(-1),体外培养的家蚕造血器官功能下降;浓度达到0.128 mmol·L~(-1)时,造血器官完全失去血球分泌能力,并逐渐死亡。
结论 初步证实2,4-DCP和NP对家蚕具有环境激素作用,并可能对其它鳞翅目昆虫具有类似的影响。
OBJECT: To investigate the effect of environment hormone on lepidopteron, studied of 2,4-Dichlorophenol (2,4-DCP) and P-nony-phenol (NP) act on the Bombyx silkworm in vivo, BmN cells, Sf9 cells and silkworm hemopoietic organ (HeO) in vitro. METHODS: Feed the Bombyx siliworm larvae with artificial diet which added the 2,4-DCP and NP, then survey its weight, growth speed, metamorphosis, bionergy and fecundity in strict conditions. Cultured the BmN and Sf9 cells include HeO with 2,4-DCP or NP-added TC-100 medium in vitro, then survey the DNA damage of BmN and the function of HeO, research the cell multiplication of BmN and Sf9 with method of 3-[45-dimethyl (thiazol-l-2-yl)]-2,5adiphenyltetrazolium bromide (MTT). RESULTS: The growth speed, weight, developmental uniformity, bionergy and fecuncity of the Bombyx silkworm descent dramatically as they eat the feed that the concentration of 2,4-DCP over 0.8 mmol L~-1 or NP exceed 0.500 mmol L~-1. But the effect on reproductive capacity vary with gender difference, it's more obvious on the late-stage of development and periods of metamorphosis such as ecolysis, pupation, edosion. There may be cumulative effect inside the Bombyx siliworm. The concentration level of 2,4-DCP and NP in the body and nature environment also have similar functions of environment hormone to silkworm. There have differences in the effect of 2,4-DCP and NP on cell multiplication of BmN and Sf9, so dose the concentration. The IC50 value of BmN and Sf9 cells to 2,4-DCP is about 0.14 mmol·L~-1, to NP about 0.05 mmol·L~-1. If the concentration of NP exceeded 0.128 mmol·L~-1, it can induce the DNA of BmN breakage, nevertheless, it also have concentration effect. The function of the HeO will decline as NP over 0.032 mml·L~-1, as come to 0.128 mmol·L~-1, it lost hemopoietic function completely, only go to death gradually.CONCLUSION: The 2,4-DCP and NP have effects of environment hormone function on Bombyx silkworm, moreover, have the similar influence to other lepidopters.
方法 通过添加含有环境激素2,4-DCP和NP的人工饲料来饲养家蚕,系统调查家蚕的生长速度、生长量、变态、生命力和生殖情况。TC-100培养基中添加2,4-DCP或NP,体外培养BmN、Sf9细胞和家蚕造血器官,进行培养BmN细胞的DNA损伤和造血器官功能调查,用MTT法调查细胞增殖。
结果 家蚕幼虫食下含0.8 mmol·L~(-1)以上浓度的2,4-DCP或0.500 mmol·L~(-1)以上浓度的NP饲料,生长速度、生长量、发育整齐度、生命力和生殖能力都明显下降;生殖能力所受影响应性别而不同;家蚕发育后期和蜕皮、化蛹、羽化等变态时期影响更加明显,在蚕体内可能有累积作用。体内和被污染后环境中的浓度水平2,4-DCP和NP对家蚕具有环境激素作用。
2,4-DCP和NP对BmN或Sf9细胞增殖的影响有一定差异,不同浓度的影响也不同;2种细胞对2,4-DCP的IC_(50)约为0.14 mmol·L~(-1),对NP约为0.05 mmol·L~(-1)。0.128mmol·L~(-1)以上浓度的NP,能诱导家蚕BmN细胞DNA链发生断裂,导致DNA损伤,而且具有较明显的浓度效应。NP的浓度超过0.032 mmol·L~(-1),体外培养的家蚕造血器官功能下降;浓度达到0.128 mmol·L~(-1)时,造血器官完全失去血球分泌能力,并逐渐死亡。
结论 初步证实2,4-DCP和NP对家蚕具有环境激素作用,并可能对其它鳞翅目昆虫具有类似的影响。
OBJECT: To investigate the effect of environment hormone on lepidopteron, studied of 2,4-Dichlorophenol (2,4-DCP) and P-nony-phenol (NP) act on the Bombyx silkworm in vivo, BmN cells, Sf9 cells and silkworm hemopoietic organ (HeO) in vitro. METHODS: Feed the Bombyx siliworm larvae with artificial diet which added the 2,4-DCP and NP, then survey its weight, growth speed, metamorphosis, bionergy and fecundity in strict conditions. Cultured the BmN and Sf9 cells include HeO with 2,4-DCP or NP-added TC-100 medium in vitro, then survey the DNA damage of BmN and the function of HeO, research the cell multiplication of BmN and Sf9 with method of 3-[45-dimethyl (thiazol-l-2-yl)]-2,5adiphenyltetrazolium bromide (MTT). RESULTS: The growth speed, weight, developmental uniformity, bionergy and fecuncity of the Bombyx silkworm descent dramatically as they eat the feed that the concentration of 2,4-DCP over 0.8 mmol L~-1 or NP exceed 0.500 mmol L~-1. But the effect on reproductive capacity vary with gender difference, it's more obvious on the late-stage of development and periods of metamorphosis such as ecolysis, pupation, edosion. There may be cumulative effect inside the Bombyx siliworm. The concentration level of 2,4-DCP and NP in the body and nature environment also have similar functions of environment hormone to silkworm. There have differences in the effect of 2,4-DCP and NP on cell multiplication of BmN and Sf9, so dose the concentration. The IC50 value of BmN and Sf9 cells to 2,4-DCP is about 0.14 mmol·L~-1, to NP about 0.05 mmol·L~-1. If the concentration of NP exceeded 0.128 mmol·L~-1, it can induce the DNA of BmN breakage, nevertheless, it also have concentration effect. The function of the HeO will decline as NP over 0.032 mml·L~-1, as come to 0.128 mmol·L~-1, it lost hemopoietic function completely, only go to death gradually.CONCLUSION: The 2,4-DCP and NP have effects of environment hormone function on Bombyx silkworm, moreover, have the similar influence to other lepidopters.
引文
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