Time-dependent mRNA expression of selected pro-inflammatory factors in the endometrium of primiparous cows postpartum

Reproductive Biology and Endocrinology - Tập 8 - Trang 1-9 - 2010
Christoph Gabler1, Claudia Fischer1, Marc Drillich2,3, Ralf Einspanier1, Wolfgang Heuwieser2
1Institute of Veterinary Biochemistry, Freie Universität Berlin, Berlin, Germany
2Clinic for Animal Reproduction, Freie Universität Berlin, Berlin, Germany
3Clinic for Ruminants, Section for Herd Health Management, Vetmeduni Vienna, Wien, Austria

Tóm tắt

Inflammatory processes and infections of the uterine wall must be accepted as a physiological event in dairy cows after calving. This might result in clinical or subclinical endometritis which is assumed to impair reproductive performance in the current lactation. Several cytokines and acute phase proteins have been discussed as local and systemic mediators of these inflammatory processes. The aim of the present study was to investigate the endometrial mRNA expression of the chemokine CXC ligand 5 (CXCL5), interleukin 1β (IL1B), IL6, IL8, tumour necrosis factor alpha (TNF), prostaglandin-endoperoxide synthase 2 (PTGS2) and haptoglobin (HP) in the postpartum period. Endometrial samples were obtained from primiparous cows (n = 5) on days 10, 17, 24, 31, 38 and 45 postpartum (pp) using the cytobrush technique. Cytological smears were prepared from cytobrush samples to determine the proportion of polymorphonuclear neutrophils (PMN). Total RNA was extracted from endometrial samples, and real-time RT-PCR was performed. A time-dependent mRNA expression of the investigated factors was found for the course of the postpartum period. In detail, a significantly higher expression of these factors was observed on day 17 pp compared to day 31 pp. Furthermore, the proportion of PMN peaked between days 10-24 pp and decreased thereafter to low percentages (< 5%) on day 31 pp and thereafter. In addition, CXCL5, IL1B, IL8 and HP mRNA expression correlated significantly with the proportion of PMN (P < 0.05). A significantly higher CXCL5, IL1B, IL6, IL8, PTGS2 and TNF mRNA content was observed in samples from cows with an inflamed endometrium compared with samples from cows with a healthy endometrium (P < 0.05). These results show that inflammatory cytokines and acute phase proteins are expressed in the bovine endometrium in a time-related manner during the postpartum period, with a significant expression peak on day 17 pp as a possible mucosal immune response in the uterus. The evaluation of the expression patterns of such candidate genes may reveal more information than only determining the percentage of PMN to judge the severity of an inflammation.

Tài liệu tham khảo

Singh J, Murray RD, Mshelia G, Woldehiwet Z: The immune status of the bovine uterus during the peripartum period. Vet J. 2008, 175: 301-309. 10.1016/j.tvjl.2007.02.003. Sheldon IM, Lewis GS, Leblanc S, Gilbert RO: Defining postpartum uterine disease in cattle. Theriogenology. 2006, 65: 1516-1530. 10.1016/j.theriogenology.2005.08.021. Gilbert RO, Shin ST, Guard CL, Erb HN, Frajblat M: Prevalence of endometritis and its effects on reproductive performance of dairy cows. Theriogenology. 2005, 64: 1879-1888. 10.1016/j.theriogenology.2005.04.022. Kasimanickam R, Duffield TF, Foster RA, Gartley CJ, Leslie KE, Walton JS, Johnson WH: Endometrial cytology and ultrasonography for the detection of subclinical endometritis in postpartum dairy cows. Theriogenology. 2004, 62: 9-23. 10.1016/j.theriogenology.2003.03.001. LeBlanc SJ, Duffield TF, Leslie KE, Bateman KG, Keefe GP, Walton JS, Johnson WH: Defining and diagnosing postpartum clinical endometritis and its impact on reproductive performance in dairy cows. J Dairy Sci. 2002, 85: 2223-2236. 10.3168/jds.S0022-0302(02)74302-6. Davies D, Meade KG, Herath S, Eckersall PD, Gonzalez D, White JO, Conlan RS, O'Farrelly C, Sheldon IM: Toll-like receptor and antimicrobial peptide expression in the bovine endometrium. Reprod Biol Endocrinol. 2008, 6: 53-10.1186/1477-7827-6-53. Fischer C, Drillich M, Odau S, Heuwieser W, Einspanier R, Gabler C: Selected pro-inflammatory factor transcripts in bovine endometrial epithelial cells are regulated during the oestrous cycle and elevated in case of subclinical or clinical endometritis. Reprod Fertil Dev. 2010, 22: 818-829. 10.1071/RD09120. Sheldon IM, Cronin J, Goetze L, Donofrio G, Schuberth HJ: Defining postpartum uterine disease and the mechanisms of infection and immunity in the female reproductive tract in cattle. Biol Reprod. 2009, 81: 1025-1032. 10.1095/biolreprod.109.077370. Kasimanickam R, Duffield TF, Foster RA, Gartley CJ, Leslie KE, Walton JS, Johnson WH: A comparison of the cytobrush and uterine lavage techniques to evaluate endometrial cytology in clinically normal postpartum dairy cows. Can Vet J. 2005, 46: 255-259. Gabler C, Drillich M, Fischer C, Holder C, Heuwieser W, Einspanier R: Endometrial expression of selected transcripts involved in prostaglandin synthesis in cows with endometritis. Theriogenology. 2009, 71: 993-1004. 10.1016/j.theriogenology.2008.11.009. Odau S, Gabler C, Holder C, Einspanier R: Differential expression of cyclooxygenase 1 and cyclooxygenase 2 in the bovine oviduct. J Endocrinol. 2006, 191: 263-274. 10.1677/joe.1.06761. LeBlanc SJ, Duffield TF, Leslie KE, Bateman KG, Keefe GP, Walton JS, Johnson WH: The effect of treatment of clinical endometritis on reproductive performance in dairy cows. J Dairy Sci. 2002, 85: 2237-2249. 10.3168/jds.S0022-0302(02)74303-8. Gautam G, Nakao T, Koike K, Long ST, Yusuf M, Ranasinghe RM, Hayashi A: Spontaneous recovery or persistence of postpartum endometritis and risk factors for its persistence in Holstein cows. Theriogenology. 2009, 73: 168-179. 10.1016/j.theriogenology.2009.08.010. Bondurant RH: Inflammation in the bovine female reproductive tract. J Anim Sci. 1999, 77 (Suppl 2): 101-110. Barlund CS, Carruthers TD, Waldner CL, Palmer CW: A comparison of diagnostic techniques for postpartum endometritis in dairy cattle. Theriogenology. 2008, 69: 714-723. 10.1016/j.theriogenology.2007.12.005. Galvao KN, Frajblat M, Brittin SB, Butler WR, Guard CL, Gilbert RO: Effect of prostaglandin F2alpha on subclinical endometritis and fertility in dairy cows. J Dairy Sci. 2009, 92: 4906-4913. 10.3168/jds.2008-1984. Gijsbers K, Gouwy M, Struyf S, Wuyts A, Proost P, Opdenakker G, Penninckx F, Ectors N, Geboes K, Van Damme J: GCP-2/CXCL6 synergizes with other endothelial cell-derived chemokines in neutrophil mobilization and is associated with angiogenesis in gastrointestinal tumors. Exp Cell Res. 2005, 303: 331-342. 10.1016/j.yexcr.2004.09.027. Santos RL, Zhang S, Tsolis RM, Baumler AJ, Adams LG: Morphologic and molecular characterization of Salmonella typhimurium infection in neonatal calves. Vet Pathol. 2002, 39: 200-215. 10.1354/vp.39-2-200. Rudack C, Jorg S, Sachse F: Biologically active neutrophil chemokine pattern in tonsillitis. Clin Exp Immunol. 2004, 135: 511-518. 10.1111/j.1365-2249.2003.02390.x. Bannerman DD, Paape MJ, Lee JW, Zhao X, Hope JC, Rainard P: Escherichia coli and Staphylococcus aureus elicit differential innate immune responses following intramammary infection. Clin Diagn Lab Immunol. 2004, 11: 463-472. Van Damme J, Wuyts A, Froyen G, Van Coillie E, Struyf S, Billiau A, Proost P, Wang JM, Opdenakker G: Granulocyte chemotactic protein-2 and related CXC chemokines: from gene regulation to receptor usage. J Leukoc Biol. 1997, 62: 563-569. Hoch RC, Schraufstatter IU, Cochrane CG: In vivo, in vitro, and molecular aspects of interleukin-8 and the interleukin-8 receptors. J Lab Clin Med. 1996, 128: 134-145. 10.1016/S0022-2143(96)90005-0. Williams MA, Cave CM, Quaid G, Solomkin JS: Chemokine regulation of neutrophil function in surgical inflammation. Arch Surg. 1999, 134: 1360-1366. 10.1001/archsurg.134.12.1360. Zerbe H, Schuberth HJ, Engelke F, Frank J, Klug E, Leibold W: Development and comparison of in vivo and in vitro models for endometritis in cows and mares. Theriogenology. 2003, 60: 209-223. 10.1016/S0093-691X(02)01376-6. Butt BM, Senger PL, Widders PR: Neutrophil migration into the bovine uterine lumen following intrauterine inoculation with killed Haemophilus somnus. J Reprod Fertil. 1991, 93: 341-345. 10.1530/jrf.0.0930341. Cobb SP, Watson ED: Immunohistochemical study of immune cells in the bovine endometrium at different stages of the oestrous cycle. Res Vet Sci. 1995, 59: 238-241. 10.1016/0034-5288(95)90010-1. Murakami S, Miyamoto Y, Skarzynski DJ, Okuda K: Effects of tumor necrosis factor-alpha on secretion of prostaglandins E2 and F2alpha in bovine endometrium throughout the estrous cycle. Theriogenology. 2001, 55: 1667-1678. 10.1016/S0093-691X(01)00511-8. Herath S, Lilly ST, Santos NR, Gilbert RO, Goetze L, Bryant CE, White JO, Cronin J, Sheldon IM: Expression of genes associated with immunity in the endometrium of cattle with disparate postpartum uterine disease and fertility. Reprod Biol Endocrinol. 2009, 7: 55-10.1186/1477-7827-7-55. Konnai S, Usui T, Ohashi K, Onuma M: The rapid quantitative analysis of bovine cytokine genes by real-time RT-PCR. Vet Microbiol. 2003, 94: 283-294. 10.1016/S0378-1135(03)00119-6. Wijayagunawardane MP, Gabler C, Killian G, Miyamoto A: Tumor necrosis factor alpha in the bovine oviduct during the estrous cycle: messenger RNA expression and effect on secretion of prostaglandins, endothelin-1, and angiotensin II. Biol Reprod. 2003, 69: 1341-1346. 10.1095/biolreprod.103.017327.