Expression of Apoptosis-related Proteins in Placenta Tissue of Yak (Bos grunniens)
MA Yue1, FAN Jiang-Feng1,*, He Hong-Hong2, HAN Xiao-Hong1, YU Si-Jiu2, CUI Yan2, XU Geng-Quan2, WANG Li-Bin1, PAN Yang-Yang2, YU Yi-Teng1, ZHAO Sheng-Xian1, MA Jin-Biao1
1 College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070, China; 2 Gansu Cattle and Sheep Embryo Engineering Technology Research Center, Lanzhou 730070, China
Abstract：Placenta is the bond between the mother and the fetus during pregnancy, During pregnancy, it undergoes a dynamic process from formation, development to maturity and degradation, and its abnormal function may lead to abnormal fetal development or even abortion. Apoptosis is an important way for placental tissue to maintain homeostasis and function properly. Studying the dynamic changes and molecular mechanism of apoptosis in placenta tissue at various stages of yak (Bos grunniens)'s pregnancy. In this study, the placental tissue of calves at various stages of pregnancy were collected to detect the expression of apoptosis-related proteins Fas (factor associated suicide)/Fasl (factor associated suicide ligand), Bcl-2 (b-cell lymphoma-2)/Bax (b-cell lymphoma-2 assaciated X protein) and Caspase3 (cysteinyl aspartate specific proteinase3)/Caspase9 (cysteinyl aspartate specific proteinase 9) by immunohistochemistry. It was shown that Fas and Fasl protein were highly expressed in the placenta and early pregnancy, and showed a downward trend as the pregnancy progresses. Bcl-2 protein was mainly expressed in fetal villus epithelium and gradually decreasing with prolonged expression during pregnancy. Bax was highly expressed in fetal villus epithelium and early trophoblasts, it had a marked increase in late pregnancy. Caspase3 was mainly expressed in the endometrium, while Caspase9 was clearly expressed in the fetal trophoblast. Evidently apoptosis-related proteins Fas/Fasl, Bcl-2/Bax and Caspase3/Caspase9 had extensive cell-specific and time-specific expression patterns in placental tissues of yak during pregnancy. It was speculated that apoptosis-associated protein-mediated apoptosis would play a important role in the normal function of placental tissue during the dynamic changes of placental tissue formation, development, maturation and degeneration. This experiment provides a theoretical basis for studying and solving the problems in the production process of yak, such as abortion, fetal growth restriction and other anomalies.
[1] 樊江峰. 2012. 正常妊娠和分娩及药物流产牦牛胎盘组织的细胞凋亡及其调节机制研究[D]. 博士学位论文, 兰州: 甘肃农业大学, 导师: 余四九. pp. 5. (Fan J F. 2012. Apoptosis and regulation mechanism of placental tissue in normal pregnancy and childbirth and medical abortion [D]. Thesis for Ph.D., Gansu Agricultural University, Suppervisor: Yu S J. pp. 5.) [2] 高丽萍, 孙丽洲, 隋灿烂, 等. 2008. Caspase3、Bcl-2 蛋白在胎盘组织中的表达及其与羊水过少的相关性研究[J]. 徐州医学院学报, (09): 607-610. (Gao L P, Sun L Z, Sui C L, et al. Expression of Caspase3 and Bcl-2 protein in placenta tissue and its correlation with amniotic fluid[J]. Journal of Xuzhou Medical College, (09): 607-610.) [3] 韩小红, 樊江峰, 于艺腾, 等. 2018. 妊娠期和分娩前后牦牛胎盘组织形态学变化及细胞凋亡的研究[J]. 农业生物技术学报, 26(10): 1714-1722. (Han X H, Fan J F, Yu Y T, et al.2018. Morphological changes and apoptosis of placenta in yak during gestation and before and after de- livery[J]. Journal of Agricultural Biotechnology, 26(10): 1714-1722.) [4] 何玉林, 郭晓青, 冯继红. 2009. Fas/FasL、Bcl-2/Bax 蛋白在母胎界面的表达[J]. 细胞与分子免疫学杂志, 25(6): 550-552. (He Y L, Guo X Q, Feng J H.2009. Expression of Fas/FasL and Bcl-2/Bax proteins at the maternal-fetal interface[J]. Journal of Cellular and Molecular Immunology, 25(6): 550-552.) [5] 姬秋梅. 2001. 中国牦牛品种资源的研究进展[J]. 自然资源报,(06): 564-568+570-569. [6] 金峰, 乔宠, 尚涛. 2011. 胎儿生长受限患者血清和胎盘的Fas 与FasL 蛋白变化及意义[J]. 中华生殖与避孕杂志, 31(3): 205-207. (Jin F, Qiao C, Shang T.2011. Changes of Fas and FasL proteins in serum and placenta of patients with fetal growth restriction[J]. Chinese Journal of Reproduction and Contraception, 31(3): 205-207.) [7] 孙莉, 陈琳, 马继红. 2017. 妊娠早期复发性流产患者妊娠组织增殖与凋亡的研究[J]. 临床误诊误治,(1): 94- 97. (Sun L, Chen L, Ma J H. 2017. Study on proliferation and apoptosis of pregnancy in patients with recurrent abortion in early pregnancy[J]. Misdiagnosis and Misdiagnosis, 2017(1): 94-97.) [8] 司超. 2018. 乙肝病毒x 蛋白结合蛋白(HBXIP)和caspase 9 蛋白在肝细胞癌中的表达及意义[D]. 硕士学位论文, 西南医科大学, 导师: 姚辉华, pp. 12. (Si C. 2018. Expression and significance of hepatitis B virus x protein binding protein (HBXIP) and caspase 9 protein in hepatocellular carcinoma[D]. Thesis for M.S., Zhangzhou: Southwestern Medical University, Suppervisor: Yao H H, pp. 12.) [9] 王豫黔, 杨鹃, 张锚链, 等. 2007. 表皮生长因子与胎儿生长受限胎盘组织Bcl-2, Bax 基因表达相关性研究[J]. 中华妇幼临床医学杂志(电子版), 3(1): 23-25. (Wang Y Q, Yang J, Zhang M L, et al.2007. Relationship between epidermal growth factor and Bcl-2, Bax gene expression in placental tissues of fetal growth restriction[J]. Chinese Journal of Women and Child Clinical Medicine, 3(1): 23-25.) [10] 字向东, 钟金城. 牦牛繁殖科学研究进展[J]. 西南民族大学学报(自然科学版), 2007, 33(1):83-87. (Zi X D, Zhong J C.Prog-ress in the research of yak breeding[J]. Journal of South-west University For Nationalities: Natural Science Edition, 2007, 33(1):83-87.) [11] 张容昶. 1989. 中国的牦牛[M]. 兰州: 甘肃科学技术出版社. pp. 224. (Zhang R C. 1989. Chinese Yak[M]. Lanzhou: Gansu Sci- ence and Technology Press., Lanzhou, China,pp. 224) [12] 赵兴绪. 2002. 兽医产科学[M]. 中国农业出版社, pp. 130 (Zhao X X. Veterinary Obstetrics[M]. China Agriculture Press. Beijing, China, pp. 130) [13] 赵海, 陈明飞, 钱宁, 等. 2017. Caspase-3 参与调节孕鼠中后期胎盘细胞凋亡的机制研究[J]. 黑龙江畜牧兽医, (8): 234-236. (Zhao H, Chen M F, Qian N, et al.2017. Mech- anism of Caspase-3 involved in regulating apoptosis of placental cells in the middle and late stages of pregnant mice[J]. Heilongjiang Animal Husbandry and Veterinary Medicine, 2017(8): 234-236.) [14] 朱国萍, 娄阳, 徐冲. 2001. Bcl-2 蛋白家族与细胞凋亡[J]. 中国细胞生物学学报, 23(1): 20-23. (Zhu G P, Lou Y, Xu C.2001. Bcl-2 protein family and apoptosis[J]. Chinese Journal of Cell Biology, 23(1): 20-23.) [15] Atia T A.2017. Placental apoptosis in recurrent miscarriage[J]. Kaohsiung Journal of Medical Sciences, 33(9): 449. [16] Abrahams V M, Straszewski Chavez S L, Guller S, et al.2004. First trimester trophoblast cells secrete Fas ligand which induces immune cell apoptosis[J]. Molecular Human Reproduction, 10(1): 55-63. [17] Adams J M, Cory S.1998. The Bcl-2 protein family: Arbiters of cell survival[J]. Science, 281(5381): 1322-1326. [18] Aschkenazi S, Straszewski S, Verwer K M, et al.2002. Differential regulation and function of the Fas/Fas ligand system in human trophoblast cells[J]. Biology of Reproduction, 66(6):1853-1861. [19] Bellgrau D, Gold D, Selawry H, et al.1995. A role for CD95 ligand in preventing graft rejection[J]. Nature, 377(6550): 630-632. [20] De Falco M, Penta R, Laforgia V, et al.2005. Apoptosis and hu- man placenta: expression of proteins belonging to dif- ferent apoptotic pathways during pregnancy[J]. Journal of experimental ＆ Clinical Cancer Research, 24(1): 25-33. [21] Drachenberg C B, Papadimitriou J C, BalatonA J, et al.2001. The total test approach to standardization of immunohis- tochemistry[J]. Archives of Pathology and Laboratory Medicine, 125(4): 471. [22] Griffith T S, Brunner T, Fletcher S M, et al.1995. Fas ligand- induced apoptosis as a mechanism of immune privilege[J]. Science, 270(5239): 1189-1192. [23] Gross A, Yin X M, Wang K, et al.1999. Caspase cleaved BID targets mitochondria and is required for cytochrome c re- lease, while BCL-XL prevents this release but not tumor necrosis factor-R1/ Fas death[J]. Journal of Biological Chemistry, 274(2): 1156-1163. [24] Hammer A and Dohr G.2000. Expression of Fas-ligand in first trimester and term human placental villi[J]. Journal Of Reproductive Immunology, 46(2):83-90. [25] Huppertz B, Kadyrov M, Kingdom J C.2006. Apoptosis and its role in the trophoblast[J]. American Journal of Obstetrics and Gynecology, 195(1): 29-39. [26] Kauma S W, Huff T F, Hayes N, et al.1999. Placental Fas ligand expression is a mechanism for maternal immune tolerance to the fetus[J]. Journal of Clinical Endocrinology ＆ Metabolism,84(6): 2188-2194. [27] Kerr J F R, Wyllie A H, Currie A R.1972. Apoptosis: A basic biological phenomenon with wideranging implications in tissue kinetics[J]. British Journal of Cancer, 26(4): 239-257. [28] Klisch K, Leiser R.2003. In bovine binucleate trophoblast giant cells, regnancy-associated glycoproteins and placental prolactin-related protein-I are conjugated to asparagine-linked N-acetylgalactosaminyl glycans[J]. Histochemistry and Cell Biology, 119(3): 211-217. [29] Nagata S.Fas and Fas ligand: a death factor and its receptor[J].Advances In Immunology:Immunopathogenesis of Type 1 Dlabetes Mellitus,Vol 100 1994; 57: 129-135. [30] Pongcharoen S, Searle R F, Bulmer J N.2004. Placental Fas and Fas ligand expression in normal early, term and molar pregnancy[J]. Placenta, 25(4): 321-330. [31] Sobrido-Cameán D, Barreiro-Iglesias A.2018. Role of caspase-8 and Fas in cell death after spinal cord injury[J]. Frontiers in molecular neuroscience, 11: 101. [32] Song J, Sapi E, Brown W, et al.2000. Expression of Fas and Fas ligand during pregnancy, lactation and involution and its potential role during mammary gland remodeling[J]. Journal of Clinical Investigation, 106: 1209-1224. [33] Ushizawa K, Takahashi T, Kaneyama K, et al.2006. Cloning of the bovine antiapoptotic regulator, Bcl2-related protein a1, and its expression in trophoblastic binucleate cells of bovine placenta[J]. Biology of Reproduction, 74(2): 344-351. [34] Wooding F B, Roberts R M, Green J A.2005. Light and elec- tron microscope immunocytochemical studies of the dis- tribution of pregnancy associated glycoproteins ( PAGs) throughout pregnancy in the cow: Possible functional implications[J]. Placenta, 26(10): 807-827. [35] Yang J, Liu X, Bhalla K, et al.1997. Prevention of apoptosis by Bcl-2: Release of cytochrome c from mitochondria blocked.[J]. Science, 1997, 275(5303): 1129-1132. [36] Yu S J, Huang Y M, Chen B X.1993. Reproductive patterns of the yak. i. reproductive phenomena of the female yak[J]. The British Veterinary Journal, 149(6): 579-583. [37] Zou H, Li Y, Liu X, et al.1999. An Apaf-1. cytochrome c multimeric complex is a functional apoptosome that activates pro Caspase-9[J]. Journal of Biological Inorganic Chemistry, 274(17): 11549-5610.
