二硫化碳通过改变卵巢黄体巨噬细胞极化和功能导致胚胎植入障碍
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摘要
目的探讨二硫化碳(CS2)暴露对孕鼠卵巢黄体巨噬细胞极化和功能的影响,从免疫损伤角度解析巨噬细胞在CS2致胚胎植入障碍中的作用。方法建立围植入期不同时间点暴露于CS2的小鼠模型,3个染毒组分别于受孕第3(GD3)、4(GD4)和5(GD5)天经腹腔染毒CS2(0.4 LD50,631.4 mg/kg)1次,各组设立平行溶剂对照(等体积橄榄油)。各组动物染毒后于不同时间点设立连续观察终点,取卵巢组织进行HE染色、免疫组织化学染色及实时荧光定量PCR实验,检测孕鼠卵巢组织中巨噬细胞极化和功能的改变。结果 (1)卵巢黄体巨噬细胞极化平衡在CS2染毒后表现为M1型巨噬细胞为主导(M1>M2,如GD3染毒组GD4观察终点,t=10.000,P<0.001),后期转变为M2型巨噬细胞为主导(M1 Objective To investigate the polarization and functional changes of ovarian corpus luteum macrophages in pregnant mice exposed to carbon disulfide( CS2) and to explore the role of macrophages in implantation failure induced by CS2 from the perspective of immune injury. Methods The animal model of exposure to CS2 at different time points was established. Single dose of CS2( 0.4 LD50,631.4 mg/kg) was injected intraperitoneally at the third( GD3),fourth( GD4),and fifth( GD5) day of gestation,and the solvent control group was set up( equal volume of olive oil). The continuous observation endpoints were designed at different time points after exposure. Ovarian tissues were collected for HE staining,immunohistochemistry,and real-time quantitative PCR detection,and the polarization and functional changes of macrophages were also determined. Results(1) The polarization balance of ovarian corpus luteum macrophages changed from the M1 type of macrophages( M1 > M2,such as GD3-treated group at GD4 observation endpoint,t =10.000,P<0.001) to the M2 type of macrophages( M1
引文
[1]王姝婷.植入期CS2暴露致孕鼠自噬水平变化及膳食补充NCG对妊娠结局的影响[D].济南:山东大学,2016.WANG Shuting.The change of autophagy in mice after exposed to carbon disulfide at peri-implantation and the effect on the pregnancy outcome after mice diet supplement NCG[D].Jinan:Shandong University,2016.
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[4]李本庆,刘军,姜万桐,等.二硫化碳对女工生殖功能影响的调查[J].中国妇幼保健,1992,7(3):51-52.LI Benqing,LIU Jun,JIANG Wantong,et al.Investigation on the effect of carbon disulfide on reproductive function of female w orkers[J].Maternal and Child Health Care of China,1992,7(3):51-52.
[5]Pieleszek A.The effect of carbon disulphide on menopause,concentration of monoamines,gonadotropins,estrogens and androgens in w omen[J].Ann Acad Med Stetin,1997,43:255-267.
[6]王志萍.二硫化碳对作业女工早期妊娠影响及其作用机理的研究[D].济南:山东大学,2005.
[7]王志萍,侯光萍,李佩贤,等.二硫化碳对作业女工受孕时间的影响[J].中华劳动卫生职业病杂志,1997,15(5):34-36.WANG Zhiping,HOU Guangping,LI Peixian,et al.Time-to-pregnancy among w omen occupationally exposed to carbon disulfide[J].Chin J Ind Hyg Occup Dis,1997,15(5):34-36.
[8]Gnainsky Y,Dekel N,Granot I.Implantation:mutual activity of sex steroid hormones and the immune system guarantee the maternal-embryo interaction[J].Semin Reprod Med,2014,32(5):337-345.
[9]Care AS,Diener KR,Jasper MJ,et al.Macrophages regulate corpus luteum development during embryo implantation in mice[J].J Clin Invest,2013,123(8):3472-3487.
[10]Wang S,Sun Y,Wu Y,et al.Down-regulation of uterine LIF expression induced by the hormonal level disorder causes embryo implantation loss after mice exposed to carbon disulfide at peri-implantation[J].Biochem Biophys Res Commun,2015,467(1):7-13.
[11]张炳珍,吴艳玲,代炳芹,等.胚胎植入期二硫化碳染毒对孕鼠血清雌激素及子宫雌激素受体-α表达的影响[J].中华劳动卫生职业病杂志,2013,31(2):88-91.ZHANG Bingzhen,WU Yanling,DAI Bingqin,et al.Effects of carbon disulfide exposure during peri-implantation on estrogen receptor-αexpression in uterus and serum level of estrogen in pregnant mice[J].Chin J Ind Hyg Occup Dis,2013,31(2):88-91.
[12]杨柳,王志萍.围植入期暴露二硫化碳对小鼠子宫组织降钙素和黄体酮水平的影响[J].中华劳动卫生职业病杂志,2013,31(12):881-885.YANG Liu,WANG Zhiping.Effect of carbon disulfide exposure during peri-implantation period on levels of calcitonin and progesterone in uterus of mice[J].Chin JInd Hyg Occup Dis,2013,31(12):881-885.
[13]Azim HA Jr,Peccatori FA,Brohée S,et al.RANK-ligand(RANKL)expression in young breast cancer patients and during pregnancy[J].Breast Cancer Res,2015,17(1):24.doi:10.1186/s13058-015-0538-7.
[14]Miyamoto A,Shirasuna K,Shimizu T,et al.Impact of angiogenic and innate immune systems on the corpus luteum function during its formation and maintenance in ruminants[J].Reprod Biol,2013,13(4):272-278.
[15]Meidan R,Klipper E,Zalman Y,et al.The role of hypoxia-induced genes in ovarian angiogenesis[J].Reprod Fertil Dev,2013,25(2):343-350.
[16]Bae WY,Choi JS,Kim JE,et al.Cinnamic aldehyde suppresses hypoxia-induced angiogenesis via inhibition of hypoxia-inducible factor-1αexpression during tumor progression[J].Biochem Pharmacol,2015,98(1):41-50.
[17]Neeman M,Abramovitch R,Schiffenbauer YS,et al.Regulation of angiogenesis by hypoxic stress:from solid tumours to the ovarian follicle[J].Int J Exp Pathol,1997,78(2):57-70.
[18]Chen MC,Lee CF,Huang WH,et al.Magnolol suppresses hypoxia-induced angiogenesis via,inhibition of HIF-1α/VEGF signaling pathw ay in human bladder cancer cells[J].Biochem Pharmacol,2013,85(9):1278-1287.
[19]Zhang Y,Xu Y,Ma J,et al.Adrenomedullin promotes angiogenesis in epithelial ovarian cancer through upregulating hypoxia-inducible factor-1αand vascular endothelial grow th factor[J].Sci Rep,2017,7:40524.doi:10.1038/srep40524.
[20]Bishop CV,Xu F,Steinbach R,et al.Changes in immune cell distribution and their cytokine/chemokine production during regression of the rhesus macaque corpus luteum[J].Biol Reprod,2017,96(6):1210-1220.
[21]Riabov V,Gudima A,Wang N,et al.Role of tumor associated macrophages in tumor angiogenesis and lymphangiogenesis[J].Front Physiol,2014,5:75.doi:10.3389/fphys.2014.00075.e Collection 2014.
[22]Ribatti D.Mast cells and macrophages exert beneficial and detrimental effects on tumor progression and angiogenesis[J].Immunol Lett,2013,152(2):83-88.
[23]陈芯仪,孙瑞雪,张文燕,等.替代活化型巨噬细胞和多发性骨髓瘤早期治疗反应的关系及机制研究[J].中华血液学杂志,2017,38(6):480-486.CHEN Xinyi,SUN Ruixue,ZHANG Wenyan,et al.Molecular mechanisms and relationship of M2-polarized macrophages w ith early response in multiple myeloma[J].Chin J Hematol,2017,38(6):480-486.
[24]Badham HJ,Winn LM.In utero and in vitro effects of benzene and its metabolites on erythroiddifferentiation and the role of reactive oxygen species[J].Toxicol Appl Pharmacol,2010,244(3):273-279.
[25]Zhang B,Shen C,Yang L,et al.DNA damage and apoptosis of endometrial cells cause loss of the early embryo in mice exposed to carbon disulfide[J].Toxicol Appl Pharmacol,2013,273(2):381-389.
[26]Wan J,Winn LM.In utero,exposure to benzene increases embryonic c-Myb and Pim-1 protein levels in CD-1 mice[J].Toxicol Appl Pharmacol,2008,228(3):326-333.
[2]Sun Y,Dai B,Yang L,et al.Carbon disulfide exposure at peri-implantation disrupts embryo implantation by decreasing integrinβ3 expression in the uterine tissue of pregnant mice[J].Chem Biol Interact,2013,206(2):126-133.
[3]保毓书,蔡世雄,赵树芬,等.二硫化碳对女工生殖结局影响的研究[J].中华劳动卫生职业病杂志,1992,(1):3-7,63-64.BAO Yushu,CAI Shixiong,ZHAO Shufen,et al.Study on the effect of carbon disulfide on the reproductive outcome of female w orkers[J].Chin J Ind Hyg Occup Dis,1992,10(1):3-7,63-64.
[4]李本庆,刘军,姜万桐,等.二硫化碳对女工生殖功能影响的调查[J].中国妇幼保健,1992,7(3):51-52.LI Benqing,LIU Jun,JIANG Wantong,et al.Investigation on the effect of carbon disulfide on reproductive function of female w orkers[J].Maternal and Child Health Care of China,1992,7(3):51-52.
[5]Pieleszek A.The effect of carbon disulphide on menopause,concentration of monoamines,gonadotropins,estrogens and androgens in w omen[J].Ann Acad Med Stetin,1997,43:255-267.
[6]王志萍.二硫化碳对作业女工早期妊娠影响及其作用机理的研究[D].济南:山东大学,2005.
[7]王志萍,侯光萍,李佩贤,等.二硫化碳对作业女工受孕时间的影响[J].中华劳动卫生职业病杂志,1997,15(5):34-36.WANG Zhiping,HOU Guangping,LI Peixian,et al.Time-to-pregnancy among w omen occupationally exposed to carbon disulfide[J].Chin J Ind Hyg Occup Dis,1997,15(5):34-36.
[8]Gnainsky Y,Dekel N,Granot I.Implantation:mutual activity of sex steroid hormones and the immune system guarantee the maternal-embryo interaction[J].Semin Reprod Med,2014,32(5):337-345.
[9]Care AS,Diener KR,Jasper MJ,et al.Macrophages regulate corpus luteum development during embryo implantation in mice[J].J Clin Invest,2013,123(8):3472-3487.
[10]Wang S,Sun Y,Wu Y,et al.Down-regulation of uterine LIF expression induced by the hormonal level disorder causes embryo implantation loss after mice exposed to carbon disulfide at peri-implantation[J].Biochem Biophys Res Commun,2015,467(1):7-13.
[11]张炳珍,吴艳玲,代炳芹,等.胚胎植入期二硫化碳染毒对孕鼠血清雌激素及子宫雌激素受体-α表达的影响[J].中华劳动卫生职业病杂志,2013,31(2):88-91.ZHANG Bingzhen,WU Yanling,DAI Bingqin,et al.Effects of carbon disulfide exposure during peri-implantation on estrogen receptor-αexpression in uterus and serum level of estrogen in pregnant mice[J].Chin J Ind Hyg Occup Dis,2013,31(2):88-91.
[12]杨柳,王志萍.围植入期暴露二硫化碳对小鼠子宫组织降钙素和黄体酮水平的影响[J].中华劳动卫生职业病杂志,2013,31(12):881-885.YANG Liu,WANG Zhiping.Effect of carbon disulfide exposure during peri-implantation period on levels of calcitonin and progesterone in uterus of mice[J].Chin JInd Hyg Occup Dis,2013,31(12):881-885.
[13]Azim HA Jr,Peccatori FA,Brohée S,et al.RANK-ligand(RANKL)expression in young breast cancer patients and during pregnancy[J].Breast Cancer Res,2015,17(1):24.doi:10.1186/s13058-015-0538-7.
[14]Miyamoto A,Shirasuna K,Shimizu T,et al.Impact of angiogenic and innate immune systems on the corpus luteum function during its formation and maintenance in ruminants[J].Reprod Biol,2013,13(4):272-278.
[15]Meidan R,Klipper E,Zalman Y,et al.The role of hypoxia-induced genes in ovarian angiogenesis[J].Reprod Fertil Dev,2013,25(2):343-350.
[16]Bae WY,Choi JS,Kim JE,et al.Cinnamic aldehyde suppresses hypoxia-induced angiogenesis via inhibition of hypoxia-inducible factor-1αexpression during tumor progression[J].Biochem Pharmacol,2015,98(1):41-50.
[17]Neeman M,Abramovitch R,Schiffenbauer YS,et al.Regulation of angiogenesis by hypoxic stress:from solid tumours to the ovarian follicle[J].Int J Exp Pathol,1997,78(2):57-70.
[18]Chen MC,Lee CF,Huang WH,et al.Magnolol suppresses hypoxia-induced angiogenesis via,inhibition of HIF-1α/VEGF signaling pathw ay in human bladder cancer cells[J].Biochem Pharmacol,2013,85(9):1278-1287.
[19]Zhang Y,Xu Y,Ma J,et al.Adrenomedullin promotes angiogenesis in epithelial ovarian cancer through upregulating hypoxia-inducible factor-1αand vascular endothelial grow th factor[J].Sci Rep,2017,7:40524.doi:10.1038/srep40524.
[20]Bishop CV,Xu F,Steinbach R,et al.Changes in immune cell distribution and their cytokine/chemokine production during regression of the rhesus macaque corpus luteum[J].Biol Reprod,2017,96(6):1210-1220.
[21]Riabov V,Gudima A,Wang N,et al.Role of tumor associated macrophages in tumor angiogenesis and lymphangiogenesis[J].Front Physiol,2014,5:75.doi:10.3389/fphys.2014.00075.e Collection 2014.
[22]Ribatti D.Mast cells and macrophages exert beneficial and detrimental effects on tumor progression and angiogenesis[J].Immunol Lett,2013,152(2):83-88.
[23]陈芯仪,孙瑞雪,张文燕,等.替代活化型巨噬细胞和多发性骨髓瘤早期治疗反应的关系及机制研究[J].中华血液学杂志,2017,38(6):480-486.CHEN Xinyi,SUN Ruixue,ZHANG Wenyan,et al.Molecular mechanisms and relationship of M2-polarized macrophages w ith early response in multiple myeloma[J].Chin J Hematol,2017,38(6):480-486.
[24]Badham HJ,Winn LM.In utero and in vitro effects of benzene and its metabolites on erythroiddifferentiation and the role of reactive oxygen species[J].Toxicol Appl Pharmacol,2010,244(3):273-279.
[25]Zhang B,Shen C,Yang L,et al.DNA damage and apoptosis of endometrial cells cause loss of the early embryo in mice exposed to carbon disulfide[J].Toxicol Appl Pharmacol,2013,273(2):381-389.
[26]Wan J,Winn LM.In utero,exposure to benzene increases embryonic c-Myb and Pim-1 protein levels in CD-1 mice[J].Toxicol Appl Pharmacol,2008,228(3):326-333.