胎衣不下奶牛母体胎盘钙离子调控机制的研究

发布时间：2018-03-03 18:02

本文选题：胎衣不下　切入点：钙离子　出处：《黑龙江八一农垦大学》2017年硕士论文　论文类型：学位论文

【摘要】：奶牛分娩结束12 h后胎衣未排出者即为胎衣不下,此病发病率高、能够继发乳房炎、蹄叶炎、子宫炎症等。为减轻奶牛业经济损失,研究者们对胎衣不下展开了大量的研究。研究者发现,饲喂含有钙离子的阴离子盐饲料可以有效降低胎衣不下的发病率。可见钙离子在生物过程调节中扮演着重要角色,它作用广泛,不但参与炎症反应、激素的调节,更重要的是它能够参与肌肉的收缩调节。因此,本实验从钙离子代谢调节机制着手,研究钙离子在奶牛母体胎盘组织细胞内的调节作用,为寻找胎衣不下发生的关键因素奠定基础,为找到治疗胎衣不下功效显著的方法提供线索。实验选取体况健康、体重、年龄、胎次接近并自然分娩的胎衣不下奶牛与胎衣正常排出奶牛(无其他疾病)各6头作为实验对象,分为胎衣不下组与对照组。采集胎衣不下组与对照组奶牛产前1~5 d及产后6 h、12 h、24 h的血液样本及母体胎盘组织,检测血浆中钙离子的浓度。运用荧光定量PCR方法检测胎衣不下奶牛与对照组奶牛母体胎盘组织中参与调节钙离子进入胞内的TRPC1、TPPV1、S100A1、SERCA、IP3R1、CACNA1D、Ry R1、PLCγ,参与钙离子流出胞外的PMCA、SLC8A1的m RNA表达量,以及调节钙离子储存的CASQ1与CALM的m RNA表达量。Western blot方法来验证部分蛋白(PLCγ、PMCA)的差异性表达与Q-PCR结果是否一致。检测调节肌肉收缩的蛋白(MLCK、MLC、Rho、Rho A)m RNA在胎衣不下与对照组奶牛的表达情况。以上实验结果显示,胎衣不下奶牛与对照组相比,产前2~5 d血浆钙离子浓度无显著差异(P0.05)产前1 d血浆钙离子浓度显著偏低(P0.05),产后6~24h血浆钙离子浓度极显著降低(P0.01)。参与调节钙离子流入细胞内的TPPV1、S100A1、SERCA、Ry R1、PLCγm RNA在胎衣不下奶牛母体胎盘组织中表达量极显著(P0.01)下调,TRPC1、IP3R1、CACNA1D m RNA表达量在胎衣不下奶牛母体胎盘组织中显著(P0.05)下调。参与调节钙离子流出胞外PMCA m RNA表达量在胎衣不下奶牛母体胎盘组织中极显著(P0.01)上调,SLC8A1 m RNA表达量在胎衣不下奶牛母体胎盘组织中显著(P0.05)上调。参与调节肌肉收缩的MLC m RNA表达量在胎衣不下奶牛母体胎盘组织中极显著(P0.01)下调,MLCK、Rho A、Rho m RNA表达量在胎衣不下奶牛母体胎盘组织中显著(P0.05)下调。说明在胎衣不下的发生过程中,以上调节钙离子进出细胞的蛋白与调控肌细胞收缩的蛋白m RNA表达发生了显著变化,通过调节母体胎盘中血管平滑肌收缩影响腺窝与绒毛的分离,影响胎衣不下的发生,并发挥重要作用。
[Abstract]:In order to alleviate the economic loss of dairy industry, the cow has a high incidence of mastitis, phlogistic, hysteritis, and so on after 12 hours of delivery, if the fetal coat is not discharged, the incidence of this disease is high, and the disease can be secondary to mastitis, phlogistic, inflammation of the uterus and so on. Researchers have carried out a lot of research on fetal coat. The researchers found that feeding anion salt diet containing calcium ions can effectively reduce the incidence of fetal coat. It is clear that calcium ion plays an important role in the regulation of biological process. It plays an important role not only in the regulation of inflammation, hormone, but also in the regulation of muscle contraction. To study the effect of calcium on the cellular regulation of placenta tissue of dairy cow, to lay a foundation for finding out the key factors of the occurrence of placenta, and to provide clues for finding the effective method of treating placenta. The age, birth order and natural delivery of the cows and the normal excretion of cows (no other diseases) 6 cows as experimental subjects, The blood samples and placenta tissue were collected from the cows of the control group and the control group on the 5th day of prepartum and the 6th hour of postpartum for 12 h and 24 h, respectively. The concentration of calcium in plasma was detected by fluorescence quantitative PCR. The expression of m RNA in placenta tissues of pregnant cows and control cows involved in regulation of calcium ions entering into the placenta was detected by means of TPPV1, S100A1, SERCACAA1, IP3R1, CACNA1D, Ry R1, PLC 纬, and the expression of m RNA in extracellular PMCACASLC8A1, which was involved in calcium exudation. Western blot method was used to verify whether the differential expression of partial protein PLC- 纬 -PMCA was consistent with Q-PCR results. The protein regulating muscle contractility was detected by MLCX, MLCX, Rho am RNA in placenta and control milk. The above results show that, Compared with the control group, There was no significant difference in plasma calcium concentration between 2 and 5 days postpartum (P0.05)) on the 1st day of prenatal, the plasma calcium concentration was significantly lower than that of P0.05, and the plasma calcium concentration at 6h postpartum significantly decreased P0.01. TPPV1 S100A1 + SERCARy R1 PLC 纬 m RNA involved in regulating the influx of calcium ions into the cells. The expression of TRPC1 / IP3R1 / CACNA1D m RNA was significantly down-regulated in placental tissues of dairy cows (P0.01). It is involved in regulating the expression of extracellular PMCA m RNA in maternal placenta. The expression of SLC8A1 m RNA was significantly up-regulated in placental tissues of dairy cows (P0.05). The expression of MLC m RNA involved in the regulation of muscle contraction was significantly increased in placenta tissues of pregnant cows (P0.01). The down-regulation of Rho Aho Rho m RNA expression in placenta tissues of dairy cows showed a significant down-regulation (P0.05), which indicated that the expression of Rho Rho m RNA in placenta of pregnant cows was significantly decreased during the development of placenta. The expression of the above proteins regulating calcium ion in and out of the cells and the protein m RNA regulating the contraction of myocytes were significantly changed. By regulating the contraction of vascular smooth muscle in the placenta of the mother, the separation of gland fossa and villi was affected, and the occurrence of fetal coat was affected. And play an important role.
【学位授予单位】：黑龙江八一农垦大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S858.23

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