胎衣不下奶牛母体胎盘中血管内皮生长因子A异常表达的分析
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摘要
目的分析胎衣不下(retained fetal membrane,RFM)奶牛母体胎盘组织中血管内皮生长因子A(vascular endothelial growth factor-A,VEGFA)的表达情况。方法提取RFM及胎衣正常排出奶牛母体胎盘组织中的总蛋白,采用荧光定量PCR及Western blot法检测两组奶牛母体胎盘组织中VEGFA的mRNA转录及蛋白表达水平。结果与胎衣正常排出组相比,RFM组奶牛母体胎盘组织中VEGFA的mRNA转录及蛋白表达水平均显著降低,差异均有统计学意义(P <0. 01)。结论发生RFM的奶牛母体胎盘组织中VEGFA表达减少,提示VEGFA可能参与该病的发生发展。
Objective To analyze the expression of vascular endothelial growth factor-A(VEGFA) in placenta of cows with retained fetal membrane(RFM). Methods Total protein was extracted from the placenta tissues of cows with and without RFM,and determined for mRNA transcription and protein expression of VEGFA by fluorescent quantitative PCR and Western blot respectively. Results Both the mRNA transcription and protein expression levels of VEGFA in placenta tissue of cows with RFM were significantly lower than those without RFM(each P < 0. 01). Conclusion The expression level of VEGFA in placenta tissue of cows with RFM decreased,indicating the possible role of VEGFA in RFM.
Objective To analyze the expression of vascular endothelial growth factor-A(VEGFA) in placenta of cows with retained fetal membrane(RFM). Methods Total protein was extracted from the placenta tissues of cows with and without RFM,and determined for mRNA transcription and protein expression of VEGFA by fluorescent quantitative PCR and Western blot respectively. Results Both the mRNA transcription and protein expression levels of VEGFA in placenta tissue of cows with RFM were significantly lower than those without RFM(each P < 0. 01). Conclusion The expression level of VEGFA in placenta tissue of cows with RFM decreased,indicating the possible role of VEGFA in RFM.
引文
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[12] ECKHARD U, HUESGEN P F, SCHILLING O, et al. Active site specificity profiling of the matrix metalloproteinase family:Proteomic identification of 4300 cleavage sites by nine MMPs explored with structural and synthetic peptide cleavage analyses[J]. Matrix Biol, 2016, 49:37-60.
[13] OLIVEIRA D M, CHALFUN-JUNIOR A, CHIZZOTTI M L, et al. Expression of genes involved in lipid metabolism in the muscle of beef cattle fed soybean or rumen-protected fat, with or without monensin supplementation[J]. J Anim Sci, 2014,92(12):5426-5436.
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[21] THOTALA D, LASZLO A, HALLAHAN D E. Cytosolic phospholipase A2 and autotaxin inhibitors as potential radiosensitizers//Phospholipases in health and disease[M]. New York:Springer New York, 2014:159-176.
[22]CHO Y C,PARK J E,PARK B C,et al. Cell cycle-dependent Cdc25C phosphatase determines cell survival by regulating apoptosis signal-regulating kinase 1[J]. Cell Death Differ,2015,22(10):1605-1617.
[23] CAO T,XING Y,YANG Y,et al. Correlation between matrix metalloproteinase expression and activation of the focal adhesion kinase signaling pathway in herpes stromal keratitis[J].Exp&Ther Med, 2014, 7(1):280-286.
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[26] GAO Y B,LI C E. Expression and clinical significance of FAK and MMP-9 in papillary thyroid carcinoma[J]. J Sichuan Ordnance, 2015, 36(6):157-160.(in Chinese)高玉宝,李常恩.甲状腺乳头状癌中FAK和MMP-9的表达及临床意义[J].兵器装备工程学报,2015, 36(6):157-160.
[27] CUEVAS B D, ABELL A N, WITOWSKY J A, et al. MEKK1regulates calpain-dependent proteolysis of focal adhesion proteins for rear-end detachment of migrating fibroblasts[J]. Embo J, 2014, 22(13):3346-3355.
[28] MARZENA L. Altered maternal serum matrix metalloproteinases MMP-2, MMP-3, MMP-9, and MMP-13 in severe early-and late-onset preeclampsia[J]. BioMed Res Int, 2017, 2017:1-9.
[29] HELSTEN T, SCHWAEDERLE M, KURZROCK R. Fibroblast growth factor receptor signaling in hereditary and neoplastic disease:biologic and clinical implications[J]. Cancer&Metast Rev, 2015, 34(3):479-496.
[30] HU Y, XU S, JIN W, et al. Effect of the PTEN gene on adhesion, invasion and metastasis of osteosarcoma cells[J]. Oncol Rep,2014, 32(4):1741-1747.
[31] HAN S, HAN L, YAO Y, et al. Activated hepatic stellate cells promote hepatocellular carcinoma cell migration and invasion via the activation of FAK-MMP9 signaling[J]. Oncol Rep,2014, 31(2):641.
[32] TORNIN J, HERMIDAPRADO F, PADDA R S, et al. FUSCHOP promotes invasion in myxoid liposarcoma through a SRC/FAK/RHO/ROCK-dependent pathway[J]. Neoplasia, 2018,20(1):44-56.
[33] ZHU Z G, WANG L, ZHANG J Y,et al. Application of analysis of principal component and cluster in blood biochemical of cows with retained fetal membranes[J]. Acta Agriculturae Boreali-occidentalis Sinica, 2017, 26(5):685-692.(in Chinese)朱永刚,王磊,张景艳,等.主成分分析法与聚类分析法在胎衣不下奶牛血液生化研究中的应用[J].西北农业学报,2017, 26(5):685-692.
[34] PALOMARES R A, GUTIERREZ J C, PORTILLO G, et al.Oxytocin treatment immediately after calving does not reduce the incidence of retained fetal membranes or improve reproductive performance in crossbred Zebu cows[J]. Theriogenology,2010, 74(8):1414-1419.
[35] BAKAKOS P, PATENTALAKIS G, PAPI A. Vascular biomarkers in asthma and COPD[J]. Curr Top Med Chem, 2016, 16(14):1599-1609.
[36] WANG J S, HAN Z X. Mechanisms and research progress of antiangiogenesis cancer therapy induced hypertention[J]. J Modern Oncol, 2018(5):800-804.(in Chinese)王金爽,韩正祥.抗血管生成靶向药物治疗肿瘤引起高血压的机制及研究进展[J].现代肿瘤医学,2018(5):800-804.
[37] YANG X Q, TAN S. Research progress of vascular endothelial growth factor in pancreatic carcinoma[J]. Hainan Med J,2014(8):1157-1160.(in Chinese)杨小秋,谈顺.血管内皮生长因子研究进展及其与胰腺癌的相关研究[J].海南医学,2014(8):1157-1160.
[38] BORO P, KUMARESAN A, PATHAK R, et al. Alteration in peripheral blood concentration of certain pro-inflammatory cytokines in cows developing retention of fetal membranes[J].Animal Reprod Sci, 2015, 157:11-16.
[39] HANSEN T D, KOEPSEL J T, LE N N, et al. Biomaterial arrays with defined adhesion ligand densities and matrix stiffness identify distinct phenotypes for tumorigenic and nontumorigenic human mesenchymal cell types[J]. Biomater Sci, 2014,2(5):745-756.
[40] ITO S. Role of RhoA/Rho-kinase and calcium sensitivity in airway smooth muscle functions//Calcium signaling in airway smooth muscle cells[M]. Cham:Springer International Publishing, 2014:285-307.
[41] MAENG J, SHE VERDIN V, SHIN H, et al. Up-regulation of Rhoa/Rho kinase pathway by translationally controlled tumor protein in vascular smooth muscle cells[J]. Int J Mol Sci,2014, 15(6):10365-10376.
[42] LIU Y, ZHENG C Y, LUO C H, et al. Analysis and validation of the differentially expressed Rho A/Rho of cow with retained fetal membranes[J]. J Heilongjiang Bayi Agri Univ,2018,30(2):34-37.(in Chinese)刘瑶,郑程远,罗春海,等.奶牛产后胎盘组织中RhoA/Rho激酶的表达与胎衣不下的关系探讨[J].黑龙江八一农垦大学学报,2018, 30(2):34-37.
[2] ZHENG C Y, ZOU X, LIN H J, et al. miRNA-185 regulates the VEGFA signaling pathway in daily cows with retained fetal membranes[J]. Theriogenology,2018,110:116-121.
[3] ZINICOLA M, KORZEC H, TEIXEIRA A G V, et al. Effects of pegbovigrastim administration on periparturient diseases, milk production, and reproductive performance of Holstein cows[J].J Dairy Sci, 2018, 101(12):11199-11217.
[4] HOOSHMANDABBASI R, ZERBE H, BAUERSACHS S, et al.Pregnancy-associated glycoproteins in cows with retained fetal membranes[J].Theriogenology,2018,105:158-163.
[5] RAPACZ-LEONARD A, RAS A, CALKA J, et al. Expression of oxytocin receptors is greatly reduced in the placenta of heavy mares with retained fetal membranes due to secondary uterine atony[J]. Equine Vet J, 2015, 47(5):623-626.
[6] WANG G J,YANG X, HAO M. Research progress of vascular endothelial growth fact or-A in the pathogenesis of cervical cancer[J]. J Int Obstetrics&Gynecol, 2016, 43(5):519-523.(in Chinese)王广娇,杨欣,郝敏.血管内皮生长因子A在宫颈癌发病机制中的研究进展[J].国际妇产科学杂志,2016, 43(5):519-523.
[7] BORRAS D, PERALES-PUCHALT A, RUIZ SACEDON N, et al. Angiogenic growth factors in maternal and fetal serum in pregnancies complicated with intrauterine growth restriction[J]. J Obstetrics&Gynaecol, 2014, 34(3):218-220.
[8] DONG H C, ZHAI X F. The research progress on the mechanism of tumor angiogenesis-related signal proteins[J]. J Modern Oncol, 2014, 22(9):2231-2233.(in Chinese)董宏超,翟校枫.VEGFA-VEGFR2相关信号蛋白作用机制的研究进展[J].现代肿瘤医学,2014, 22(9):2231-2233.
[9] EGOROVA A, PETROSYAN M, MARETINA M, et al. Anti-angiogenic treatment of endometriosis via anti-VEGFA siRNA delivery by means of peptide-based carrier in a rat subcutaneous model[J]. Gene Ther, 2018, 25(8):548-555.
[10] DAS V, KC R, LI X, et al. Blockade of vascular endothelial growth factor receptor-1(Flt-1), reveals a novel analgesic for osteoarthritis-induced joint pain[J]. Gene Rep,2018,11:94-100.
[11] ADEYINKA F D, LAVEN R A, DE NICOLO G, et al. The use of stereology method to estimate the volume of feto-maternal exchange area of the bovine placentome during gestation[J].Anatomical Record, 2016, 299(11):1571-1577.
[12] ECKHARD U, HUESGEN P F, SCHILLING O, et al. Active site specificity profiling of the matrix metalloproteinase family:Proteomic identification of 4300 cleavage sites by nine MMPs explored with structural and synthetic peptide cleavage analyses[J]. Matrix Biol, 2016, 49:37-60.
[13] OLIVEIRA D M, CHALFUN-JUNIOR A, CHIZZOTTI M L, et al. Expression of genes involved in lipid metabolism in the muscle of beef cattle fed soybean or rumen-protected fat, with or without monensin supplementation[J]. J Anim Sci, 2014,92(12):5426-5436.
[14] GONG J, ZHU S, ZHANG Y, et al. Interplay of VEGFa and MMP2 regulates invasion of glioblastoma[J].Tumor Biol,2014, 35(12):11879-11885.
[15] SZEBENYI S A, OGURA T, SATHYANESAN A, et al. Increases in intracellular calcium via activation of potentially multiple phospholipase C isozymes in mouse olfactory neurons[J].Front Cell Neurosci, 2014, 8:336.
[16] BERNAL-QUIROS M, WU Y Y, ALARCON-RIQUELME M E,et al. BANK1 and BLK act through phospholipase C gamma2 in B-cell signaling[J]. PLoS One, 2013, 8(3):e59842.
[17] SEDIGHI O, MAKHLOUGH A, SHOKRZADEH M, et al. Association between plasma selenium and glutathione peroxidase levels and severity of diabetic nephropathy in patients with type two diabetes mellitus[J]. Nephrourol Mon, 2014, 6(5):e21355.
[18] ANISHKIN A, LOUKIN S H, TENG J, et al. Feeling the hidden mechanical forces in lipid bilayer is an original sense[J].Proc Natl Acad Sci USA, 2014, 111(22):7898-7905.
[19] HAYES T K, DER C J. Targeting the Raf-MEK-ERK mitogenactivated protein kinase cascade for the treatment of RAS,mutant cancers//Ras superfamily small C proteins:biology and mechanisms[M]. Vienna:Springer Vienna,2014:135-156.
[20] HENG Y J, LIONG S, PERMEZEL M, et al. Human cervicovaginal fluid biomarkers to predict term and preterm labor[J].Frontiers Physiol, 2015, 6:151.
[21] THOTALA D, LASZLO A, HALLAHAN D E. Cytosolic phospholipase A2 and autotaxin inhibitors as potential radiosensitizers//Phospholipases in health and disease[M]. New York:Springer New York, 2014:159-176.
[22]CHO Y C,PARK J E,PARK B C,et al. Cell cycle-dependent Cdc25C phosphatase determines cell survival by regulating apoptosis signal-regulating kinase 1[J]. Cell Death Differ,2015,22(10):1605-1617.
[23] CAO T,XING Y,YANG Y,et al. Correlation between matrix metalloproteinase expression and activation of the focal adhesion kinase signaling pathway in herpes stromal keratitis[J].Exp&Ther Med, 2014, 7(1):280-286.
[24] MTSUGAKI A, ARAMOTO G, NINOMIYA T, et al. Abnormal arrangement of a collagen/apatite extracellular matrix orthogonal to osteoblast alignment is constructed by a nanoscale periodic surface structure[J]. Biomaterials, 2015, 37:134-143.
[25] MUSIAL K, ZWOLIfiSKA D. Pleiotropic functions of TIMP-1 in patients with chronic kidney disease[J]. Cell&Mol Life Sci,2014, 71(8):1547-1548.
[26] GAO Y B,LI C E. Expression and clinical significance of FAK and MMP-9 in papillary thyroid carcinoma[J]. J Sichuan Ordnance, 2015, 36(6):157-160.(in Chinese)高玉宝,李常恩.甲状腺乳头状癌中FAK和MMP-9的表达及临床意义[J].兵器装备工程学报,2015, 36(6):157-160.
[27] CUEVAS B D, ABELL A N, WITOWSKY J A, et al. MEKK1regulates calpain-dependent proteolysis of focal adhesion proteins for rear-end detachment of migrating fibroblasts[J]. Embo J, 2014, 22(13):3346-3355.
[28] MARZENA L. Altered maternal serum matrix metalloproteinases MMP-2, MMP-3, MMP-9, and MMP-13 in severe early-and late-onset preeclampsia[J]. BioMed Res Int, 2017, 2017:1-9.
[29] HELSTEN T, SCHWAEDERLE M, KURZROCK R. Fibroblast growth factor receptor signaling in hereditary and neoplastic disease:biologic and clinical implications[J]. Cancer&Metast Rev, 2015, 34(3):479-496.
[30] HU Y, XU S, JIN W, et al. Effect of the PTEN gene on adhesion, invasion and metastasis of osteosarcoma cells[J]. Oncol Rep,2014, 32(4):1741-1747.
[31] HAN S, HAN L, YAO Y, et al. Activated hepatic stellate cells promote hepatocellular carcinoma cell migration and invasion via the activation of FAK-MMP9 signaling[J]. Oncol Rep,2014, 31(2):641.
[32] TORNIN J, HERMIDAPRADO F, PADDA R S, et al. FUSCHOP promotes invasion in myxoid liposarcoma through a SRC/FAK/RHO/ROCK-dependent pathway[J]. Neoplasia, 2018,20(1):44-56.
[33] ZHU Z G, WANG L, ZHANG J Y,et al. Application of analysis of principal component and cluster in blood biochemical of cows with retained fetal membranes[J]. Acta Agriculturae Boreali-occidentalis Sinica, 2017, 26(5):685-692.(in Chinese)朱永刚,王磊,张景艳,等.主成分分析法与聚类分析法在胎衣不下奶牛血液生化研究中的应用[J].西北农业学报,2017, 26(5):685-692.
[34] PALOMARES R A, GUTIERREZ J C, PORTILLO G, et al.Oxytocin treatment immediately after calving does not reduce the incidence of retained fetal membranes or improve reproductive performance in crossbred Zebu cows[J]. Theriogenology,2010, 74(8):1414-1419.
[35] BAKAKOS P, PATENTALAKIS G, PAPI A. Vascular biomarkers in asthma and COPD[J]. Curr Top Med Chem, 2016, 16(14):1599-1609.
[36] WANG J S, HAN Z X. Mechanisms and research progress of antiangiogenesis cancer therapy induced hypertention[J]. J Modern Oncol, 2018(5):800-804.(in Chinese)王金爽,韩正祥.抗血管生成靶向药物治疗肿瘤引起高血压的机制及研究进展[J].现代肿瘤医学,2018(5):800-804.
[37] YANG X Q, TAN S. Research progress of vascular endothelial growth factor in pancreatic carcinoma[J]. Hainan Med J,2014(8):1157-1160.(in Chinese)杨小秋,谈顺.血管内皮生长因子研究进展及其与胰腺癌的相关研究[J].海南医学,2014(8):1157-1160.
[38] BORO P, KUMARESAN A, PATHAK R, et al. Alteration in peripheral blood concentration of certain pro-inflammatory cytokines in cows developing retention of fetal membranes[J].Animal Reprod Sci, 2015, 157:11-16.
[39] HANSEN T D, KOEPSEL J T, LE N N, et al. Biomaterial arrays with defined adhesion ligand densities and matrix stiffness identify distinct phenotypes for tumorigenic and nontumorigenic human mesenchymal cell types[J]. Biomater Sci, 2014,2(5):745-756.
[40] ITO S. Role of RhoA/Rho-kinase and calcium sensitivity in airway smooth muscle functions//Calcium signaling in airway smooth muscle cells[M]. Cham:Springer International Publishing, 2014:285-307.
[41] MAENG J, SHE VERDIN V, SHIN H, et al. Up-regulation of Rhoa/Rho kinase pathway by translationally controlled tumor protein in vascular smooth muscle cells[J]. Int J Mol Sci,2014, 15(6):10365-10376.
[42] LIU Y, ZHENG C Y, LUO C H, et al. Analysis and validation of the differentially expressed Rho A/Rho of cow with retained fetal membranes[J]. J Heilongjiang Bayi Agri Univ,2018,30(2):34-37.(in Chinese)刘瑶,郑程远,罗春海,等.奶牛产后胎盘组织中RhoA/Rho激酶的表达与胎衣不下的关系探讨[J].黑龙江八一农垦大学学报,2018, 30(2):34-37.