[논문]Pro-inflammatory Cytokines and Their Receptors: Expression and Regulation in the Uterine Endometrium during the Estrous Cycle in Pigs

Yoo, Inkyu; Kim, Minjeong; Han, Jisoo; Jang, Hwanhee; Choi, Sun-Ho; Ka, Hakhyun

doi:10.12750/jet.2016.31.4.323

Pro-inflammatory Cytokines and Their Receptors: Expression and Regulation in the Uterine Endometrium during the Estrous Cycle in Pigs 원문보기

Journal of embryo transfer = 한국수정란이식학회지, v.31 no.4, 2016년, pp.323 - 333

Yoo, Inkyu (Department of Biological Science and Technology, Yonsei University) , Kim, Minjeong (Department of Biological Science and Technology, Yonsei University) , Han, Jisoo (Department of Biological Science and Technology, Yonsei University) , Jang, Hwanhee (Department of Biological Science and Technology, Yonsei University) , Choi, Sun-Ho (Swine Science Division, National Institute of Animal Science, Rural Development Administration) , Ka, Hakhyun (Department of Biological Science and Technology, Yonsei University)

Abstract ▼ AI-Helper

Pro-inflammatory cytokines, interleukin-$1{\beta}$(IL1B), IL6, and tumor necrosis factor-alpha (TNF), are known to play important roles in regulating the endometrial function in the uterus during the estrous cycle and pregnancy in several species. However, the expression and function of these cytokines and their receptors in the uterine endometrium during the estrous cycle have not been studied in pigs. Thus, this study determined the expression and regulation of IL1B, IL6, TNF and their respective receptors, IL1R1, IL1RAP, IL6R, GP130, TNFRSF1A, and TNFRSF1B during the estrous cycle in pigs. To analyze levels of each gene expression in the uterine endometrium we obtained from endometrial tissues on Days 0, 3, 6, 9, 12, 15, and 18 of the estrous cycle. Real-time RT-PCR analysis showed that levels of IL1B, IL1RAP, IL6R, GP130, TNF, TNFRSF1A, and TNFRSF1B mRNAs were highest on Day 15 or 18 of the estrous cycle, which corresponds to the proestrus period. Levels of IL1R1 were highest on Day 0, while levels of IL6 were biphasic with high levels on Day 6 and Day 15. The abundance of IL1B, IL6, IL6R, and TNF mRNAs was decreased by progesterone, while levels of GP130 were increased by progesterone in endometrial tissue explants. These results showed that expression of pro-inflammatory cytokines and their receptors changed stage-specifically during the estrous cycle and regulated by progesterone in the uterine endometrium in pigs, suggesting that these pro-inflammatory cytokines may be involved in the regulation endometrial function during the estrous cycle in pigs.

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가설 설정

In this study, our results showed that the expression of most IL1B, IL6, TNF and their receptors was changed in the uterine endometrium with higher levels at proestrus of the estrous cycle, which corresponds to the follicular phase of the ovarian cycle. The expression pattern of the pro-inflammatory cytokines and their receptors in this study made us to hypothesize that the transition of dominance of steroid hormones estrogen and progesterone during the estrous cycle is a critical factor for expression of the pro-inflammatory cytokines and their receptors. Thus, we took advantage of the explant cultures using uterine endometrial tissues from immature gilts to examine the effect of steroid hormones on cytokine expression and found that increasing doses of P₄decreased the levels of IL1B, IL6, IL6R, and TNF mRNAs and increased GP130 mRNA levels.

제안 방법

Data from real-time RT-PCR for IL1B, IL6, TNF and their receptor expression during the estrous cycle and from explant cultures were subjected to regression analysis using the General Linear Models procedures of SAS (Cary, NC). As sources of variation, the model included day to evaluate steady-state levels of IL1B, IL6, TNF and their receptor mRNAs during the estrous cycle and dose to evaluate effects of steroid hormones in explant cultures. Data are presented as means with SEM.
In this study we evaluated the expression of pro-inflammatory cytokines and their receptors in the uterine endometrium during the estrous cycle in pigs. Result of this study showed that the expression of pro-inflammatory cytokines IL1B and TNF mRNAs, but not IL6 mRNAs, changed in the uterine endometrium during the estrous cycle with higher levels at proestrus than diestrus.
Explant tissues were then harvested and total RNA was extracted for real-time RT-PCR analysis to determine expression levels for IL1B, IL6, TNF and their receptor mRNAs. These experiments were conducted using endometria from eight immature gilts.
The expression and function of IL1B, IL6, and TNF in the uterine endometrium under the normal and disease conditions have been reported in many species, but the precise role of these cytokines in the uterine endometrium during the estrous cycle has not been well understood. Thus, to initiate the study on the role of IL1B, IL6, and TNF in the uterine endometrium during the estrous cycle in pigs, this study determined the expression of pro-inflammatory cytokine IL1B, IL6, TNF and their receptors in the uterine endometrium during the estrous cycle and the effects of ovarian steroid hormones on endometrial expression of these molecules in pigs.

대상 데이터

Endometrial explants from immature gilts were cultured in DMEM/F-12 with increasing doses (0, 5, 50, 500, and 5000 pg/ml) of estradiol-17β (E₂) at 37℃ for 24 h. Experiments were conducted using endometrial tissues from eight immature gilts. Abundance of mRNA expression based on real-time RT-PCR analyses is relative to that for each mRNAs in the control group of endometrial explants after normalization of transcript amounts to RPL7, UBB and TBP mRNA.

데이터처리

Data from real-time RT-PCR for IL1B, IL6, TNF and their receptor expression during the estrous cycle and from explant cultures were subjected to regression analysis using the General Linear Models procedures of SAS (Cary, NC). As sources of variation, the model included day to evaluate steady-state levels of IL1B, IL6, TNF and their receptor mRNAs during the estrous cycle and dose to evaluate effects of steroid hormones in explant cultures.

이론/모형

The results are reported as expression relative to the level detected on Day 12 of the estrous cycle after normalization of the transcript amount to the endogenous RPL7, UBB and TBP control by the 2-ΔΔCT method (Livak and Schmittgen 2001).

To analyze levels of IL1B, IL6, TNF and their receptor mRNAs in the uterine endometrial tissues, real-time RT-PCR was performed using the Applied Biosystems StepOnePlus System (Applied Biosystems, Foster City, CA) using the SYBR Green method. Complementary DNAs were synthesized from 4 µg total RNA isolated from different uterine endometrial tissues, and newly synthesized cDNAs (total volume of 21 μl) were diluted 1:4 with nuclease-free water and then used for PCR.

성능/효과

In conclusion, this study showed that pro-inflammatory cytokine IL1B, IL6, TNF, and their receptors are expressed in the uterine endometrium with high levels during proestrus of the estrous cycle and progesterone decreases the expression of some of cytokines in pigs. These results indicate that the expression of endometrial pro-inflammatory cytokines is regulated depending the stage of the estrous cycle and pro-inflammatory cytokines may play an important role in regulation of endometrial function during the estrous cycle in pigs.

In this study, our results showed that the expression of most IL1B, IL6, TNF and their receptors was changed in the uterine endometrium with higher levels at proestrus of the estrous cycle, which corresponds to the follicular phase of the ovarian cycle. The expression pattern of the pro-inflammatory cytokines and their receptors in this study made us to hypothesize that the transition of dominance of steroid hormones estrogen and progesterone during the estrous cycle is a critical factor for expression of the pro-inflammatory cytokines and their receptors.

Although the expression and function of pro-inflammatory cytokines at the maternal-fetal interface have been reported, their function in the uterine endometrium during the estrous cycle is very limited. In this study, the finding that several pro-inflammatory cytokines and their receptor were highly expressed in the uterine endometrium at late diestrus and proestrus indicates that these cytokines may play a critical role in endometrial remodeling during this period. In primates, dramatic endometrial remodeling occurs at menstruation and is associated with TNF.

In this study we evaluated the expression of pro-inflammatory cytokines and their receptors in the uterine endometrium during the estrous cycle in pigs. Result of this study showed that the expression of pro-inflammatory cytokines IL1B and TNF mRNAs, but not IL6 mRNAs, changed in the uterine endometrium during the estrous cycle with higher levels at proestrus than diestrus. Levels of cytokine receptors IL1RAP, IL6R, GP130, TNFRSF1A, and TNFRSF1B mRNAs were also highest at proestrus phase during the estrous cycle.

However, estrogen did not have any effect on IL1B, IL6, TNF and their receptors in explant cultures. These data suggest that the endometrial expression of IL1B, IL6, IL6R, and TNF during the estrous cycle is regulated by progesterone rather than estrogen; during metestrus to diestrus, the progesterone-dominant period, progesterone down-regulates IL1B, IL6, IL6R, and TNF expression, while progesterone-mediated down-regulation of IL1B, IL6, IL6R, and TNF expression during late diestrus to proestrus is released due to the decreased levels of progesterone during this period. In pigs, infiltration and distribution of leukocytes in the uterine endometrium change during the estrous cycle depending on the levels of steroid hormones with the high number of lymphocytes and macrophages at diestrus and neutrophils at proestrus (Kaeoket et al.

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참고문헌 (50)

Anegon I, Cuturi MC, Godard A, Moreau M, Terqui M, Martinat-Botte F and Soulillou JP. 1994. Presence of leukaemia inhibitory factor and interleukin 6 in porcine uterine secretions prior to conceptus attachment. Cytokine 6(5):493-499.

상세보기

Blitek A, Morawska E and Ziecik AJ. 2012. Regulation of expression and role of leukemia inhibitory factor and interleukin-6 in the uterus of early pregnant pigs. Theriogenology 78(5):951-964.

상세보기

Chen HL, Yang YP, Hu XL, Yelavarthi KK, Fishback JL and Hunt JS. 1991. Tumor necrosis factor alpha mRNA and protein are present in human placental and uterine cells at early and late stages of gestation. Am. J. Pathol. 139(2): 327-335.

상세보기

Choudhuri R and Wood GW. 1993. Production of interleukin-1, interleukin-6, and tumor necrosis factor alpha in the uterus of pseudopregnant mice. Biol. Reprod. 49(3):596-603.

상세보기

Clark AR and Kruger JA. 2016. Mathematical modeling of the female reproductive system: from oocyte to delivery. Wiley Interdiscip. Rev. Syst. Biol. Med.

Correia-Alvarez E, Gomez E, Martin D, Carrocera S, Perez S, Peynot N, Giraud-Delville C, Caamano JN, Balseiro A, Sandra O, Duranthon V and Munoz M. 2015. Early embryonic and endometrial regulation of tumor necrosis factor and tumor necrosis factor receptor 2 in the cattle uterus. Theriogenology 83(6):1028-1037.

상세보기

De M, Sanford TH and Wood GW. 1992. Detection of Interleukin-1, Interleukin-6, and Tumor-Necrosis-Factor-Alpha in the Uterus during the 2nd-Half of Pregnancy in the Mouse. Endocrinology 131(1):14-20.

Diehl S and Rincon M. 2002. The two faces of IL-6 on Th1/Th2 differentiation. Mol. Immunol. 39(9):531-536.

상세보기

Franczak A, Wojciechowicz B, Zmijewska A, Kolakowska J and Kotwica G. 2013. The effect of interleukin 1beta and interleukin 6 on estradiol-17beta secretion in the endometrium of pig during early pregnancy and the estrous cycle. Theriogenology 80(2):90-98.

상세보기

Franczak A, Zmijewska A, Kurowicka B, Wojciechowicz B, Petroff BK and Kotwica G. 2012. The effect of tumor necrosis factor alpha (TNFalpha), interleukin 1beta (IL1beta) and interleukin 6 (IL6) on endometrial PGF2alpha synthesis, metabolism and release in early-pregnant pigs. Theriogenology 77(1):155-165.

상세보기

Fumuso E, Giguere S, Wade J, Rogan D, Videla-Dorna I and Bowden RA. 2003. Endometrial IL-1beta, IL-6 and TNF-alpha, mRNA expression in mares resistant or susceptible to post-breeding endometritis. Effects of estrous cycle, artificial insemination and immunomodulation. Vet. Immunol. Immunopathol. 96(1-2):31-41.

상세보기

Gray CA, Bartol FF, Tarleton BJ, Wiley AA, Johnson GA, Bazer FW and Spencer TE. 2001. Developmental biology of uterine glands. Biol. Reprod. 65(5):1311-1323.

상세보기

Haider S and Knofler M. 2009. Human tumour necrosis factor: physiological and pathological roles in placenta and endometrium. Placenta 30(2):111-123.

상세보기

Healy LL, Cronin JG and Sheldon IM. 2015. Polarized Epithelial Cells Secrete Interleukin 6 Apically in the Bovine Endometrium. Biol. Reprod. 92(6):151.

상세보기

Hirano T, Akira S, Taga T and Kishimoto T. 1990. Biological and clinical aspects of interleukin 6. Immunol. Today 11(12):443-449.

상세보기

Hondo E, Kokubu K, Kato K and Kiso Y. 2005. Localization of interieukin-6 receptor rnRNA in the pregnant and non-pregnant mouse uterus. J. Reprod. Develop. 51(6):777-781.

상세보기

Hunt JS, Chen HL, Hu XL and Tabibzadeh S. 1992. Tumor necrosis factor-alpha messenger ribonucleic acid and protein in human endometrium. Biol. Reprod. 47(1):141-147.

상세보기

Hunt JS, Miller L, Roby KF, Huang J, Platt JS and DeBrot BL. 1997. Female steroid hormones regulate production of pro-inflammatory molecules in uterine leukocytes. J. Reprod. Immunol. 35(2):87-99.

상세보기

Hunter CA and Jones SA. 2015. IL-6 as a keystone cytokine in health and disease. Nat. Immunol. 16(5):448-457.

상세보기

Jacobs AL, Sehgal PB, Julian J and Carson DD. 1992. Secretion and hormonal regulation of interleukin-6 production by mouse uterine stromal and polarized epithelial cells cultured in vitro. Endocrinology 131(3):1037-1046.

상세보기

Jana B, Kozlowska A, Andronowska A and Jedlinska-Krakowska M. 2008. The effect of tumor necrosis factor-alpha (TNF-alpha), interleukin (IL)-1 beta and IL-6 on chorioamnion secretion of prostaglandins (PG)F 2 alpha and E2 in pigs. Reprod. Biol. 8(1):57-68.

상세보기

Ka H, Jaeger LA, Johnson GA, Spencer TE and Bazer FW. 2001. Keratinocyte growth factor is up-regulated by estrogen in the porcine uterine endometrium and functions in trophectoderm cell proliferation and differentiation. Endocrinology 142(6):2303-2310.

상세보기

Kaeoket K, Persson E and Dalin AM. 2002. Corrigendum to "The sow endometrium at different stages of the oestrus cycle: studies on morphological changes and infiltration by cells of the immune system." [Anim. Reprod. Sci. 65 (2001) 95-114]. Anim. Reprod. Sci. 73(1-2):89-107.

상세보기

Kelly RW. 2002. Inflammatory mediators and cervical ripening. J. Reprod. Immunol. 57(1-2):217-224.

상세보기

Laird SM, Tuckerman EM, Cork BA and Li TC. 2000. Expression of nuclear factor kappa B in human endometrium; role in the control of interleukin 6 and leukaemia inhibitory factor production. Mol. Hum. Reprod. 6(1):34-40.

상세보기

Lim KJ, Odukoya OA, Ajjan RA, Li TC, Weetman AP and Cooke ID. 1998. Profile of cytokine mRNA expression in peri-implantation human endometrium. Mol. Hum. Reprod. 4(1):77-81.

상세보기

Livak KJ and Schmittgen TD. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 25(4):402-408.

상세보기

Mathew DJ, Newsom EM, Guyton JM, Tuggle CK, Geisert RD and Lucy MC. 2015. Activation of the transcription factor nuclear factor-kappa B in uterine luminal epithelial cells by interleukin 1 Beta 2: a novel interleukin 1 expressed by the elongating pig conceptus. Biol. Reprod. 92(4):107.

상세보기

Mathialagan N, Bixby JA and Roberts RM. 1992. Expression of interleukin-6 in porcine, ovine, and bovine preimplantation conceptuses. Mol. Reprod. Dev. 32(4):324-330.

상세보기

Modric T, Kowalski AA, Green ML, Simmen RC and Simmen FA. 2000. Pregnancy-dependent expression of leukaemia inhibitory factor (LIF), LIF receptor-beta and interleukin-6 (IL-6) messenger ribonucleic acids in the porcine female reproductive tract. Placenta 21(4):345-353.

상세보기

Palm F, Walter I, Budik S, Kolodziejek J, Nowotny N and Aurich C. 2008. Influence of different semen extenders and seminal plasma on PMN migration and on expression of IL-1beta, IL-6, TNF-alpha and COX-2 mRNA in the equine endometrium. Theriogenology 70(5):843-851.

상세보기

Prins JR, Gomez-Lopez N and Robertson SA. 2012. Interleukin-6 in pregnancy and gestational disorders. J. Reprod. Immunol. 95(1-2):1-14.

상세보기

Robertson SA and Seamark RF. 1990. Granulocyte macrophage colony stimulating factor (GM-CSF) in the murine reproductive tract: stimulation by seminal factors. Reprod. Fertil. Dev. 2(4):359-368.

상세보기

Robertson SA, Mayrhofer G and Seamark RF. 1992. Uterine Epithelial-Cells Synthesize Granulocyte-Macrophage Colony-Stimulating Factor and Interleukin-6 in Pregnant and Nonpregnant Mice. Biol. Reprod. 46(6):1069-1079.

상세보기

Robertson SA, Christiaens I, Dorian CL, Zaragoza DB, Care AS, Banks AM and Olson DM. 2010. Interleukin-6 is an essential determinant of on-time parturition in the mouse. Endocrinology 151(8):3996-4006.

상세보기

Roby KF and Hunt JS. 1994. Mouse endometrial tumor necrosis factor-alpha messenger ribonucleic acid and protein: localization and regulation by estradiol and progesterone. Endocrinology 135(6):2780-2789.

상세보기

Ross JW, Malayer JR, Ritchey JW and Geisert RD. 2003. Characterization of the interleukin-1beta system during porcine trophoblastic elongation and early placental attachment. Biol. Reprod. 69(4):1251-1259.

상세보기

Sanford TR, De M and Wood GW. 1992. Expression of colony-stimulating factors and inflammatory cytokines in the uterus of CD1 mice during days 1 to 3 of pregnancy. J. Reprod. Fertil. 94(1):213-220.

상세보기

Seo H, Choi Y, Shim J, Choi Y and Ka H. 2012. Regulatory mechanism for expression of IL1B receptors in the uterine endometrium and effects of IL1B on prostaglandin synthetic enzymes during the implantation period in pigs. Biol. Reprod. 87(2):31.

상세보기

Seo H, Choi Y, Shim J, Yoo I and Ka H. 2014. Comprehensive analysis of prostaglandin metabolic enzyme expression during pregnancy and the characterization of AKR1B1 as a prostaglandin F synthase at the maternal-conceptus interface in pigs. Biol. Reprod. 90(5):99.

상세보기

Simon C, Gimeno MJ, Mercader A, O'Connor JE, Remohi J, Polan ML and Pellicer A. 1997. Embryonic regulation of integrins beta 3, alpha 4, and alpha 1 in human endometrial epithelial cells in vitro. J. Clin. Endocrinol. Metab. 82(8):2607-2616.

Simon C, Valbuena D, Krussel J, Bernal A, Murphy CR, Shaw T, Pellicer A and Polan ML. 1998. Interleukin-1 receptor antagonist prevents embryonic implantation by a direct effect on the endometrial epithelium. Fertil. Steril. 70(5):896-906.

상세보기

Soede NM, Langendijk P and Kemp B. 2011. Reproductive cycles in pigs. Anim. Reprod. Sci. 124(3-4):251-258.

상세보기

Spencer TE, Wiley AA and Bartol FF. 1993. Neonatal age and period of estrogen exposure affect porcine uterine growth, morphogenesis, and protein synthesis. Biol. Reprod. 48(4):741-751.

상세보기

Subramaniam S, Stansberg C and Cunningham C. 2004. The interleukin 1 receptor family. Dev. Comp. Immunol. 28(5):415-428.

상세보기

Tabibzadeh S, Kong QF, Babaknia A and May LT. 1995. Progressive rise in the expression of interleukin-6 in human endometrium during menstrual cycle is initiated during the implantation window. Hum. Reprod. 10(10):2793-2799.

상세보기

Tarleton BJ, Wiley AA and Bartol FF. 1999. Endometrial development and adenogenesis in the neonatal pig: effects of estradiol valerate and the antiestrogen ICI 182,780. Biol. Reprod. 61(1):253-263.

상세보기

Woodward EM, Christoffersen M, Campos J, Betancourt A, Horohov D, Scoggin KE, Squires EL and Troedsson MH. 2013. Endometrial inflammatory markers of the early immune response in mares susceptible or resistant to persistent breeding-induced endometritis. Reproduction 145(3):289-296.

상세보기

Yelavarthi KK, Chen HL, Yang YP, Cowley BD, Jr., Fishback JL and Hunt JS. 1991. Tumor necrosis factor-alpha mRNA and protein in rat uterine and placental cells. J. Immunol. 146(11):3840-3848.

상세보기

Zhao H, Kalish F, Schulz S, Yang Y, Wong RJ and Stevenson DK. 2015. Unique roles of infiltrating myeloid cells in the murine uterus during early to midpregnancy. J. Immunol. 194(8):3713-3722.

상세보기

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