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苏云金芽孢杆菌杀虫 Cry1ab 毒素不影响哺乳动物肠道上皮细胞的膜完整性:一项体外研究。

Bacillus thuringiensis insecticidal Cry1ab toxin does not affect the membrane integrity of the mammalian intestinal epithelial cells: An in vitro study.

作者信息

Shimada Nobuaki, Miyamoto Kazuhisa, Kanda Khozo, Murata Hideo

机构信息

National Institute of Animal Health, Tsukuba, Ibaraki 305-0856, Japan.

出版信息

In Vitro Cell Dev Biol Anim. 2006 Jan-Feb;42(1-2):45-9. doi: 10.1007/s11626-006-0011-0.

Abstract

The mammalian intestinal epithelium has been found, based on in vivo experiments, to be resistant to insecticidal Cry toxins, which are derived from Bacillus thuringiensis and fatally damage insect midgut cells. Thus, the toxins are commonly used as a genetic resource in insect-resistant transgenic plants for feed. However, Cry toxins bind to the cellular brush border membrane vesicle (BBMV) of mammalian intestinal cells. In this study, we investigated the affinity of Cry1Ab toxin, a lepidopteran-specific Cry1-type toxin, to the cellular BBMV of two mammalian intestinal cells as well as the effect of the toxin on the membrane potential of three mammalian intestinal cells compared to its effects on the silkworm midgut cell. We found that Cry1Ab toxin did bind to the bovine and porcine BBMV, but far more weakly than it did to the silkworm midgut BBMV. Furthermore, although the silkworm midgut cells developed severe membrane potential changes within 1 h following the toxin treatment at a final concentration of 2 mug/ml, no such membraneous changes were observed on the bovine, porcine, and human intestinal cells. The present in vitro results suggest that, although Cry1Ab toxin may bind weakly or nonspecifically to certain BBMV components in the mammalian intestinal cell, it does not damage the cell's membrane integrity, thus exerting no subsequent adverse effects on the cell.

摘要

基于体内实验发现,哺乳动物的肠道上皮细胞对杀虫性Cry毒素具有抗性,这种毒素源自苏云金芽孢杆菌,会致命地损伤昆虫中肠细胞。因此,这些毒素通常被用作抗虫转基因饲料作物的遗传资源。然而,Cry毒素会与哺乳动物肠道细胞的细胞刷状缘膜囊泡(BBMV)结合。在本研究中,我们研究了鳞翅目特异性Cry1型毒素Cry1Ab毒素对两种哺乳动物肠道细胞的细胞BBMV的亲和力,以及该毒素对三种哺乳动物肠道细胞膜电位的影响,并将其与对家蚕中肠细胞的影响进行了比较。我们发现Cry1Ab毒素确实会与牛和猪的BBMV结合,但结合能力远弱于对家蚕中肠BBMV的结合能力。此外,尽管在家蚕中肠细胞中,在最终浓度为2微克/毫升的毒素处理后1小时内就出现了严重的膜电位变化,但在牛、猪和人的肠道细胞中未观察到此类膜变化。目前的体外实验结果表明,尽管Cry1Ab毒素可能会与哺乳动物肠道细胞中的某些BBMV成分发生弱结合或非特异性结合,但它不会破坏细胞的膜完整性,因此不会对细胞产生后续的不利影响。

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