اثر منیزیم جیره بر پارامترهای تولیدمثلی و بیان ژن‌های ماتریکس متالوپروتئیناز 2 و 9 و مهارکننده بافتی 2 در جفت گاوهای هلشتاین

نوع مقاله : مقاله پژوهشی

نویسندگان

گروه علوم دامی، دانشکده کشاورزی، دانشگاه زنجان، زنجان، ایران

چکیده

هدف از انجام این آزمایش بررسی اثر منیزیم جیره بر عملکرد تولید‌مثلی و بیان ژن‌های متالوپروتئیناز 2 (MMP-2)، متالوپروتئیناز 9  (MMP-9) و مهارکننده بافتی 2(TIMP-2) در جفت بود. تعداد 50 راس گاو در هر تیمار  از 3 هفته مانده به زایش به طور تصادفی به چهار تیمار آزمایشی اختصاص یافتند. تیمار‌ها شامل تیمار شاهد (منیزیم در سطح توصیه‌های (NRC)، تیمار سولفات ‌منیزیم، تیمار کربنات ‌منیزیم و تیمار اکسید ‌منیزیم (منیزیم 6/0 درصد ماده خشک جیره) بود. پس از زایش از جفت هر گاو نمونه‌برداری انجام شده و  بیان  نسبی ژن ها مورد بررسی قرار گرفت. نمونه‌گیری از خون پیش و پس از زایش انجام شده و متابولیت‌های پلاسما اندازه‌گیری شد. در تیمار اکسید منیزیم و سولفات منیزیم بیان ژن MMP-9 بالاتر بود و درصد جفت ماندگی در تیمارهای آزمایشی پائین‌تر از گروه شاهد بود (05/0>P ) تیمارهای آزمایشی تاثیر معنی‌داری بر بیان ژن‌هایMMP-2  و TIMP-2 نداشتند. جیره‌های آزمایشی باعث بهبود وضعیت منیزیم و انسولین پلاسما و کاهش معنی‌دار NEFA شدند (05/0>P) تیمارها تاثیری بر امتیاز برگشت رحمی، اولین فحلی و تعداد تلقیح نداشتند. تیمار اکسید منیزیم آبستنی در اولین تلقیح و آبستنی تا روز 210 پس از زایش را بهبود داد. دام ها در تیمارهای آزمایشی نسبت به گروه شاهد روزهای باز کمتری داشتند (05/0> P) نتایج نشان داد که افزایش منیزیم جیره در مقایسه با توصیه های  NRC (2001) می تواند منجر به کاهش جفت ماندگی و بهبود باروری شود.

کلیدواژه‌ها

موضوعات


عنوان مقاله [English]

Effect of dietary magnesium on reproductive parameters and expression of matrix metalloproteinase 2 and 9 and tissue inhibitor 2 in the placenta of Holstein cows

نویسندگان [English]

  • Forood Ehsanbakhsh
  • Hamid Amanlou
  • Mohammad Hossein Shahir
Department of Animal Sciences, Faculty of Agriculture, University of Zanjan, Zanjan, Iran.
چکیده [English]

The aim of this experiment was to evaluate the effect of dietary Mg on reproduction performance and expression of MMP-2, MMP-9 and TIMP-2 genes in the placenta. 50 cows in each treatments, from 3 weeks to calving were randomly assigned to four experimental treatments. Treatments included control treatment (magnesium at the level of NRC recommendations), magnesium sulfate, magnesium carbonate and magnesium oxide treatments (magnesium 0.6% of DM). Placenta samples were taken after calving and mRNA expression were investigated. Blood samples were collected pre and postpartum and plasma metabolites were measured. Relative mRNA expression of MMP-9 in magnesium oxide and magnesium sulfate treatments was higher and the percent of retained placenta in this treatments was lower than control group. Experimental treatments had no significant effect on the expression of MMP-2 and TIMP-2. Experimental diets had no effect on uterine involution score, first estrus and AI/conception. Treatments improved plasma Mg and insulin concentrations and reduced NEFA significantly. Magnesium oxide treatment improved pregnancy at first AI and pregnancy by 210 days postpartum. Cows in the experimental diets had fewer open days than the control group. The results showed that increasing dietary Mg compared to NRC (2001) recommendations could reduce retained placenta and improve fertility.

کلیدواژه‌ها [English]

  • Magnesium
  • Gene expression
  • Retained placenta
  • Pregnancy

Extended Abstract

Introduction

Maintaining health and production during transition period is an important challenge in dairy herds. Reproduction and performance of dairy cows are influenced by several factors. Metabolic diseases such as ketosis and retained placenta can have a negative effect on reproduction and cause economic losses in dairy farms. Magnesium is an important mineral in the diet. Lack of magnesium in the diet can have negative consequences on animal reproduction and health. Several studies have shown that increasing plasma magnesium reduces retained placenta and reduces first ovulation time. Higher plasma magnesium can improved conception rate. The purpose of this study was to investigate the effect of dietary magnesium and plasma magnesium level on reproductive performance and the expression of genes related to retained placenta.

 

 

 

Materials and Methods

200 multiparous Holstein cows with body weight 726± 31.2, body condition score 3.35± 0.32 were enrolled in a completely randomized design at 21 d before expected calving. Dietary treatments were control diet (CO; Mg at the level of conventional diets), magnesium sulfate diet (MgS; Mg= 0.6% of DM), magnesium carbonate diet (MgC; Mg= 0.6% of DM), and magnesium oxide diet (MgO; Mg= 0.6% of DM). All cows received the same postpartum diet.  Blood samples were collected 4 h after morning feeding from the on −21, −14, −7, −3, 1, 3, 7, 14 and 21 d relative to expected calving date. After calving, blood samples were taken weekly. Magnesium, insulin and NEFA of plasma were analyzed pre and postpartum. After calving, placenta samples were taken from cows and placed in liquid nitrogen to investigate MMP-2, MMP-9 and TIMP-2 genes expression. Uterine involution score, number of services per conception (AI/conception), first estrus time and days open were recorded for all of 200 cows.

 

Results and discussion

The results showed that pre and postpartum plasma magnesium and insulin were increased with supplementation of magnesium treatments. Magnesium oxide decreased plasma NEFA (P ≤ 0.05). Magnesium oxide decreased the percentage of retained placenta and increased the percentage of pregnancy at first AI and pregnancy up to 210 days after calving (P ≤ 0.05). Uterine involution score was not different between treatments. Experimental treatments had no significant effect on relative expression of genes MMP-2 and TIMP-2. Relative expression of MMP-9 was significantly higher in the magnesium oxide and sulfate diets than in the control group (P ≤ 0.05).

 

Conclusion

The results of this study showed that higher magnesium in diet and plasma increases magnesium and insulin of plasma and reduces blood NEFA. Also, magnesium treatments, especially magnesium oxide, reduce retained placenta and improve reproduction performance.

AOAC. (1990). Official Methods of Analysis. 15th ed. Association of Official Analytical Chemists, Arlington, VA.
Aungier, S. P., Roche, J. F., Diskin, M. G., Crowe, M. A. (2014). Risk factors that affect reproductive target achievement in fertile dairy cows. Journal of Dairy Science, 97, 3472– 3487.
Barbagallo, M., Dominguez, L. J., Galioto, A., Ferlisi, A., Cani, C., Pineo, M. A., Busardo, A & Paolisso, G. (2003). Role of magnesium in insulin action, diabetes and cardio-metabolic syndrome X. Molecular Aspects of Medicine, 24, 39-52.
Beagley, J. C., Whitman, K. J., Baptiste, K. E & Scherzer, J. (2010). Physiology and treatment of retained fetal membranes in cattle. J Vet Intern Med, 24, 261–268.
Chapinal, N., Carson, M. E., LeBlanc, S. J., Leslie, K. E., Godden, S., Capel, M., Santos, J. E., Overton, M. W. & Duffield, T. F. (2012). The association of serum metabolites in the transition period with milk production and early-lactation reproductive performance. Journal of Dairy Science, 95, 1301–1309.
De Koster, J. D. & Opsomer, G. (2013). Insulin resistance in dairy cows. Veterinary Clinics of North America: Food Animal Practice, 29, 299-322.
Dilly, M., Hambruch, N., Shenavai, S., Schuler, G., Froehlich, R., Haeger, J. D., Ozalp, G. R & Pfarrer, C. (2011). Expression of matrix metalloproteinase (MMP)-2, MMP-14 and tissue inhibitor of matrix metalloproteinase (TIMP)-2 during bovine placentation and at term with or without placental retention. Theriogenology, 75, 1104–1114.
Dubuc, J., Duffield, T. F, Leslie, K. E, Walton, J. S & S. J. LeBlanc, S. J. (2012). Risk factors and effects of postpartum anovulation in dairy cows. Journal of Dairy Science, 95, 1845–1854.
Geng, J., Huang, C & Jiang, S. (2016). Roles and Regulation of the Matrix Metalloproteinase System in Parturition. Mol. Reprod. Dev. DOI 10.1002/mrd.22626.
Grober, U., Schmidt, J. & Kisters, K. (2015). Magnesium in prevention and therapy. Nutrients, 7, 8199-8226.
Grohn, Y. T & Rajala-Schultz, P. J. (2000). Epidemiology of reproductive performance in dairy
Cows. Animal Reproduction Science, 61, 605- 614.
Goff, J. P. (2014). Calcium and magnesium disorders. Veterinary Clinics of North America: Food Animal Practice, 30, 359–381.
Jeong, J. K., Choi, I. S., Moon, S. H., Kang, H. G. & Kim I. H. (2018). Relationship between serum magnesium concentration during the transition period, peri- and postpartum disorders, and reproductive performance in dairy cows. Livestock Science, 213, 1-6.
Jittakhot S., Schonewille, J.T., Wouterse, H., Uijttewaal, A.W.J., Yuangklang, C. & Beynen, A.C.  (2004). Increasing magnesium intakes in relation to magnesium absorption in dry cows. Journal of Dairy Research, 71, 297–303.
Kizaki, K., Ushizawa, K., Takahashi, T., Yamada, O., Todorok, J., Sato, T., Ito, A & Hashizume, K. (2008). Gelatinase (MMP-2 and -9) expression profiles during gestation in the bovine endometrium. Reproductive Biology and Endocrinology, 6, 1-11.
Li, J. Y., Zhang, W. Y., Wang, X., & Zou, L.Y. (2016). Effect of magnesium sulfate on MMP-9 and AQP-9 protein in placenta of patients with hypertensive disorders complicating pregnancy. Zhonghua Yi Xue Za Zhi, 96, 2421-2423.
Leroy, J. L. M. R., Vanholder, T., Mateusen, B., Christophe, A., Opsomer, G., De Kruif, A., Genicot, G.  & Van Soom, A. (2005). Non-esterified fatty acids in follicular fluid of dairy cows and their effect on developmental capacity of bovine oocytes in vitro. Reproduction, 130, 485–495.
Lucy, M. C. (2001). Reproductive Loss in High-Producing Dairy Cattle: Where Will It End?. Journal of Dairy Science, 84, 1277- 1293.
Maj, J.G, & Kankofer. M. (1997). Activity of 72-kDa and 92-kDa matrix metalloproteinases in placental tissues of cows with and without retained fetal membranes. Placenta, 18, 683–687.
Ospina, P. A., Nydam, D. V., Stokol, T. & Overton, T. R. (2010). Associations of elevated nonesterified fatty acids and beta-hydroxybutyrate concentrations with early lactation reproductive performance and milk production in transition dairy cattle in the Northeastern United States. Journal of Dairy Science, 93, 1596–1603.
Pfaffl, M. W. (2001). A new mathematical model for relative quantification in real-time RT–PCR. Nucleic Acids Research, 29, 2002- 2007.
Qu, Y., Fadden, A. N., Traber, M. G. & Bobe, G. (2014). Potential risk indicators of retained placenta and other diseases in multiparous cows. Journal of Dairy Science, 97, 4151-65.
Rosner, J. Y., Gupta, M., McGill, M., Xue, X., Chatterjee, P. K., Yoshida-Hay, M., Robeson, W & Metz, C. N. (2016). Magnesium deficiency during pregnancy in mice impairs placental size and function. Placenta, 39, 87–93.
Thatcher, W. W., Bilby, T. R., Bartolome, J. A., Silvestre, F., Staples, C. R & Santos, J. E. P. (2006). Strategies for improving fertility in the modern dairy cow. Theriogenology, 65, 30–44.
Tsiamadis, V., Banos, G., Panousis, N., Kritsepi-Konstantinou, M., Arsenos, G & Valergakis, G.E. (2016). Genetic parameters of subclinical macromineral disorders and major clinical diseases in postparturient Holstein cows. Journal of Dairy Science, 99, 8901- 8914.
Tillard, E., Humblot, P., Faye, B., Lecomte, P., Dohoo, I & Bocquier, F. (2008). Postcalving factors affecting conception risk in Holstein dairy cows in tropical and sub-tropical conditions. Theriogenology, 69, 443–457.
Van Saun, R.J. (2014). Dairy nutrition. Veterinary Clinics of North America: Food Animal Practice, 30, 115-337.
Visse, R & Nagase, H. (2003). Matrix Metalloproteinases and Tissue Inhibitors of Metalloproteinases. Circulation Research, 92, 827-839.
Walter, I & Boos, A. (2001). Matrix Metalloproteinases (MMP-2 and MMP-9) and Tissue Inhibitor-2 of Matrix Metalloproteinases (TIMP-2) in the Placenta and Interplacental Uterine Wall in Normal Cows and in Cattle with retention of  Fetal Membranes. Placenta, 22, 473- 483.
Yue, H., Lee, J. D., Shimizu, H., Uzui, H., Mitsuke, Y & Ueda, T. (2003). Effects of magnesium on the production of extracellular matrix metalloproteinases in cultured rat vascular smooth muscle cells. Atherosclerosis, 166, 271-/277.
Yue, H., Uzui, H., Lee, J. D., Shimizu, H. & Ueda, T. (2004). Effects of magnesium on matrix metalloproteinase-2 production in cultured rat cardiac fibroblasts. Basic Res Cardiol, 99, 257- 263.