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高迁移率族蛋白B1(HMGB1)可增强在无牛血清白蛋白(BSA)条件下体外发育的猪孤雌胚胎。

High mobility group box 1 (HMGB1) enhances porcine parthenotes developing in vitro in the absence of BSA.

作者信息

Shen Xing-Hui, Jin Yong-Xun, Ko Yeoung-Gye, Chung Hak-Jae, Cui Xiang-Shun, Kim Nam-Hyung

机构信息

Department of Animal Science, Chungbuk National University, Gaesin-dong, Cheongju, Chungbuk 361-763, South Korea.

出版信息

Theriogenology. 2006 Dec;66(9):2077-83. doi: 10.1016/j.theriogenology.2006.05.019. Epub 2006 Jul 24.

Abstract

High mobility group box 1 (HMGB1) is considered a component of chromatin and membranes with a role in a variety of biologically important processes. The aim of this study was to determine the effects of HMGB1 on the viability and development of porcine diploid parthenotes cultured in vitro. In vitro derived 4-cell parthenotes were cultured to blastocysts, with or without recombinant HMGB1, in the presence or absence of BSA. The addition of 1, 10, 100 or 1000ng/mL HMGB1 into NCSU 23 medium containing 0.4% BSA did not enhance the development of 4-cell parthenotes to the blastocyst stage and did not change the total number of nuclei in the blastocysts. However, addition of 10 or 100ng/mL HMGB1 into NCSU 23 medium in the absence of BSA increased (P<0.05) both the development rate of parthenotes to the blastocyst stage and total cell numbers. When cultured in NCSU23 medium supplemented with 10 or 100ng/mL HMGB1 and without BSA, apoptosis in parthenotes at the blastocyst stage was decreased (P<0.05). Based on real time RT-PCR, the addition of HMGB1 to the culture medium in the absence of BSA decreased mRNA expression of pro-apoptotic genes Bak (P<0.005) or Caspase3 (Casp3, P<0.01), but not Bcl-xL (Bcl2l2). In conclusion, we inferred that recombinant HMGB1 in the culture medium in the absence of BSA prevented apoptosis of porcine parthenotes and enhanced porcine embryo viability.

摘要

高迁移率族蛋白B1(HMGB1)被认为是染色质和细胞膜的一个组成部分,在多种生物学重要过程中发挥作用。本研究的目的是确定HMGB1对体外培养的猪二倍体孤雌胚胎的活力和发育的影响。体外获得的4细胞期孤雌胚胎在有或无重组HMGB1、有或无牛血清白蛋白(BSA)的情况下培养至囊胚期。在含有0.4% BSA的NCSU 23培养基中添加1、10、100或1000 ng/mL HMGB1,并未提高4细胞期孤雌胚胎发育至囊胚期的比例,也未改变囊胚中的细胞核总数。然而,在无BSA的NCSU 23培养基中添加10或100 ng/mL HMGB1,孤雌胚胎发育至囊胚期的比例和总细胞数均增加(P<0.05)。当在补充有10或100 ng/mL HMGB1且无BSA的NCSU23培养基中培养时,囊胚期孤雌胚胎的凋亡减少(P<0.05)。基于实时逆转录聚合酶链反应(RT-PCR),在无BSA的情况下向培养基中添加HMGB1可降低促凋亡基因Bak(P<0.005)或半胱天冬酶3(Caspase3,P<0.01)的mRNA表达,但对Bcl-xL(Bcl2l2)无影响。总之,我们推断在无BSA的培养基中添加重组HMGB1可防止猪孤雌胚胎凋亡并提高猪胚胎活力。

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