补肾安胎方对人绒毛膜外滋养层细胞HTR-8分泌功能及HLA-G基因表达的调控作用

齐丹; 谈勇

补肾安胎方对人绒毛膜外滋养层细胞HTR-8分泌功能及HLA-G基因表达的调控作用

齐丹^1,2,
谈勇^1,

1.
南京中医药大学第一临床医学院，江苏南京 210023
2.
南京医科大学附属南京市妇幼保健院，江苏南京 210004

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出版历程
- 收稿日期: 2015-12-21
- 修回日期: 2016-02-15
- 刊出日期: 2016-03-10

Regulatory Effect of Bushen Antai Recipe on the Secretion Function and HLA-G Expression of Human Extravillous Cytotrophoblasts HTR-8

QIDan^1,2,
TANYong^1
,

1.
The First Clinical Medical College, Nanjing University of Chinese Medicine, Nanjing, 210023, China
2.
Nanjing Maternity and Child Health Care Hospital Affiliated to Nanjing Medical University, Nanjing, 210004, China

摘要

摘要: 目的观察补肾安胎方对人绒毛膜外滋养层细胞HTR-8分泌功能及人类白细胞抗原G（HLA-G）基因表达的调控，探讨其免疫保胎机制。方法雌性SD大鼠灌胃制备空白血清和补肾安胎方含药血清。将人绒毛膜外滋养层细胞HTR-8细胞分为4组：空白对照组、低剂量组（5%中药血清组）、中剂量组（10%中药血清组）、高剂量组（20%中药血清组），培养24 h后，ELISA法测定培养上清液中雌二醇（E₂）、孕激素(P）、人绒毛膜促性腺激素(HCG)及辅助性T细胞（Th）调控因子白介素-4(IL-4) 和干扰素 -γ（IFN-γ）水平，RT-PCR法测定HLA-G mRNA表达。结果与空白对照组相比，低剂量组在促进E₂、P分泌方面无统计学意义（P>0.05），中、高剂量组可有效提高细胞上清E₂、P含量，且高剂量组高于中剂量组（P<0.05）。3组含药血清均能明显提高滋养细胞HCG的分泌水平（P<0.05），3组间差异无显著性意义（P>0.05）。低剂量组即可促进Th2型细胞因子IL-4分泌，但差异无统计学意义（P>0.05），中、高剂量组可有效促进Th2型细胞因子IL-4含量，组间差异无统计学意义（P>0.05）。3组含药血清均能显著抑制Th1型细胞因子IFN-γ（P<0.05），中、高剂量组均可增加HLA-G mRMA的表达，有显著差异（P<0.05），且高剂量组高于中剂量组（P<0.05）。结论补肾安胎方含药血清可以改善滋养细胞分泌功能，使母胎免疫向Th2型转化，并可能部分通过上调HLA-G基因表达实现。
- 补肾安胎方 /
- 滋养细胞 /
- 免疫耐受 /
- HLA-G
Abstract: OBJECTIVE To study the effects of Bushen Antai Recipe (BAR) on the secretion function and HLA-G expression of human extravillous cytotrophoblasts HTR-8. METHODS SD female rats were treated with BAR or normal saline to prepare for BAR-containing serum or control serum. After centrifugation of the serum， HTR-8 were randomly divided into 4 groups： control group， low-dose BAR (BAR-L) group， medium dose BAR (BAR-M) group and high dose BAR (BAR-H) group. The BAR-L， BAR-M and BAR-H group were treated with 5%， 10% or 20% BAR-containing serum， respectively. The supematants were harvested at 24 h of the culture， and then the levels of hormone， including E₂ (estradiol)， progesterone (P) and human chorionic gonadotropin (HCG)， and regulatory factors of Helper T cells (Th)， including interleukin -4 (IL-4) and interferon gamma (IFN-γ)， were measured by ELISA in culture supernatant. The mRNA of HLA-G was analyzed using quantitative RT-PCR. RESULTS The BAR-L group could promote the secretion of E₂ and P with no statistically significant difference， the BAR-M group and BAR-H group could effectively improve the secretion of E₂ and P， and the BAR-H group is higher than the BAR-M group（P<0.05）.The three groups could improve the HCG secretion effectively（P<0.05），while there were no significant difference among them（P>0.05）. The BAR-L group could promote the secretion of IL-4 with no statistically significant difference（P>0.05）.The BAR-M group and BAR-H group could effectively improve the IL-4 secretion， while there were no significant difference between them（P>0.05）. And the three groups could down-regulate the IFN-γ secretion effectively（P<0.05）. The BAR-M group and high dose group could improve the HLA-G expression effectively（P<0.05)， and the BAR-H group is higher than the BAR-M group（P<0.05）. CONCLUSION The BAR could elevate the secretion in human extravillous cytotrophoblasts and result in the bias response for Th2 type cytokine at the maternal-fetal interface， which might be partially achieved through up-regulating HLA-G expression.
- Bushen Antai Recipe /
- human extravillous cytotrophoblasts /
- immune tolerance /
- HLA-G

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参考文献(30)

[1]	Lemaoult J， Daouya M， Wu J， et al. Synthetic HLA-G proteins for therapeutic use in transplantation[J]. FASEB J， 2013， 27(9): 3643-3651.
[2]	刘淮，黄淑晖．妊娠期免疫系统的变化和免疫耐受机制[J]．中国实用妇科与产科杂志，2010， 26（6）：465-468．
[3]	Liu H， Huang SH.Changes of the immune system and the mechanism in immune tolerance during pregnancy[J]. Chin J Pract Gynecol Obstet， 2010， 26(6): 465-468.
[4]	陈奇．中药药理研究方法学[M]．北京：人民卫生出版社，1993：1167-1170．
[5]	Chen Q. Methodology on Chinese Medicinal Pharmacology[M]. Beijing: People's medical publishing house，1993：1167-1170．
[6]	Akhter A， Das V， Naik S， et al. Upregulation of HLA-G in JEG-3 cells by dexamethasone and hydrocortisone[J]. Arch Gynecol Obstet， 2012， 285(1): 7-14.
[7]	王文娟．CD4⁺T细胞在原因不明复发性流产中的研究进展[J]．国际妇产科学杂志，2013， 40（6）：525-529．
[8]	Wang WJ. The role of CD4⁺T cells in unexplained recurrent spontaneous abortion:an update[J]. J Intern Obstet Gynecol， 2013， 40(6): 525-529.
[9]	Ivanova-Todorova E， Mourdjeva M， Kyurkchiev D， et al. HLA-G expression is up-regulated by progesterone in mesenchymal stem cells[J]. Am J Reprod Immunol， 2009， 62(1): 25-33.
[10]	岳兰，谭继英，李少红．妊娠免疫耐受机制的研究进展[J]．中国妇幼保健，2012， 27（5）：784-786．
[11]	Yue L， Tan JY， Li SH.Research progress of the mechanism in immune tolerance during pregnancy[J]. Mater Child Health Care China， 2012， 27(5): 784-786.
[12]	Mukri F， Bourne T， Bottomley C， et al. Evidence of early first-trimester growth restriction in pregnancies that subsequently end in miscarriage[J]. BJOG， 2008， 115(10): 1273-1278.
[13]	Marengoni A， Winblad B， Fratiglioni L. In utero and early-life conditions and adult health and disease[J]. N Engl J Med， 2008， 359(14): 1523; author reply 1524.
[14]	Thornton CA. Immunology of pregnancy[J]. Proceed Nutr Society， 2010， 69(3): 357-365.
[15]	Yie SM， Li LH， Li GM， et al. Progesterone enhances HLA-G gene expression in JEG-3 choriocarcinoma cells and human cytotrophoblasts in vitro[J]. Hum Reprod， 2006， 21(1): 46-51.
[16]	Graham CH， Hawley TS， Hawley RG， et al. Establishment and characterization of first trimester human trophoblast cells with extended lifespan[J]. Exp Cell Res， 1993， 206(2): 204-211.
[17]	Morrish DW，Whitley GS，Cartwright JE，et al.In vitro models to study trophoblast function and dysfunction-a workshop report[J]. Placenta，2002，(SupplA):S144-S148.
[18]	Bechi N， Sorda G， Spagnoletti A， et al. Toxicity assessment on trophoblast cells for some environment polluting chemicals and 17β-estradiol[J]. Toxicol Vitro， 2013， 27(3): 995-1000.
[19]	邢长英，归绥琪，陶敏芳，等．补肾益气方影响人早孕绒毛滋养细胞分泌功能的研究[J]．生殖与避孕，2013， 33（8）：518-523．
[20]	Xing CY， Gui SQ， Tao MF， et al. Effect of kidney-replenishing herb on the secretion of human trophoblasts[J]. Reprod Contracept， 2013， 33(8): 518-523.
[21]	张玲，王海燕，王文君，等．补肾益气方对人蜕膜CD4⁺T细胞分泌IL-4和IFN-γ的调控作用[J]．中国免疫学杂志，2009， 25（5）：421-424．
[22]	Zhang L， Wang HY， Wang WJ， et al. The formula of tonifying kidney of vital essence modulates secretion of IL-4 and IFN-γ by CD4⁺ positive T cells of human decidual[J]. Chin J Immunol， 2009， 25(5): 421-424.
[23]	王英兰，苏放明，陈德珩，等．sHLA-G对复发性流产患者滋养细胞侵袭功能的影响[J]．中山大学学报:医学科学版，2012， 33（4）：524-527．
[24]	Wang YL， Su FM， Chen DH， et al. Effect and significance of exogenous soluble HLA-G intervention on trophoblast invasion in patients of recurrent spontaneous abortion[J]. J Sun Yat-sen Univ:Med Sci， 2012， 33(4): 524-527.
[25]	王力，闻姬，韩彩艳．寿胎丸调节原因不明复发性自然流产患者免疫失衡机制[J]．北京中医药大学学报，2012， 35（11）：781-785．
[26]	Wang L， Wen J， Han CY. Reversed effect of Shoutai Wan on immunologic dissonance through regulating mononuclear granulocytes in patients with unknown recurrent spontaneous abortion (type of kidney deficiency)[J]. J Beijing Univ Tradit Chin Med， 2012， 35(11): 781-785.
[27]	马红霞，尤昭玲，刘华，等．菟丝子总黄酮对早孕期人细胞滋养细胞增殖能力的影响及其信号机制[J]．中药材，2009， 32（6）：939-943．
[28]	Ma HX， You ZL， Liu H， et al. Effect of TFCC on proliferation of the first trimester human cytotrophoblast and its possible signal transduction pathway[J]. J Chin Med Mater， 2009， 32(6): 939-943.
[29]	董川川，陈亚琼．单味中药保胎的研究现状[J]．光明中医，2013， 28（2）：403-405．
[30]	Dong CC， Chen YQ. Current research on treatment of tocolysis by single Chinese herbal medicine[J]. Guangming J Chin Med， 2013， 28(2): 403-405.