TRPV6基因沉默对蛋鸡十二指肠和空肠钙离子跨细胞转运相关蛋白表达的影响

doi:10.11843/j.issn.0366-6964.2018.11.023

畜牧兽医学报 ›› 2018, Vol. 49 ›› Issue (11): 2505-2513.doi: 10.11843/j.issn.0366-6964.2018.11.023

TRPV6基因沉默对蛋鸡十二指肠和空肠钙离子跨细胞转运相关蛋白表达的影响

孟范勇¹, 邓益锋¹, 杨俊花², 崔军¹, 周振雷¹, 侯加法^1*

1. 南京农业大学动物医学院, 南京 210095;
2. 上海市农业科学院农产品质量标准与检测技术研究所, 上海 201403

收稿日期:2018-03-12 出版日期:2018-11-23 发布日期:2018-11-23
通讯作者: 侯加法,教授,博士生导师,E-mail:houjiafa@163.com
作者简介:孟范勇(1989-),男,山东高密人,硕士生,主要从事畜禽骨骼生物学研究,E-mail:845854077@qq.com
基金资助:
国家自然科学基金（31272618；30972234）

Effects of TRPV6 Gene Silencing on the Expression of Proteins Involved in Calcium Ions Transcellular Transport in Duodenum and Jejunum of Laying Hens

MENG Fan-yong¹, DENG Yi-feng¹, YANG Jun-hua², CUI Jun¹, ZHOU Zhen-lei¹, HOU Jia-fa^1*

1. College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China;
2. Institute for Agri-food Standard and Testing Technology, Shanghai Academy of Agricultural Sciences, Shanghai 201403, China

Received:2018-03-12 Online:2018-11-23 Published:2018-11-23

摘要/Abstract

摘要：

本试验旨在研究瞬时性受体电位通道香草酸受体6（transient receptor potential vanilloid receptor 6，TRPV6）基因沉默对蛋鸡十二指肠和空肠跨细胞钙离子转运相关蛋白表达、血浆钙磷浓度、PTH水平及骨密度的影响。将64只220日龄高产蛋鸡随机分为对照组和TRPV6基因沉默组，分别一次性注射生理盐水和pSIREN-TRPV6-3质粒，连续观察28 d。结果表明，与对照组相比，pSIREN-TRPV6-3质粒注射第7、14和21天，十二指肠和空肠中均检测到ZsGreen蛋白表达；TRPV6、CaBP-D28K mRNA和蛋白表达水平均显著降低（P<0.05或P<0.01）；血浆PTH浓度显著升高（P<0.05或P<0.01）；此外，血浆钙、磷浓度不受影响，股骨和胫骨骨密度有降低趋势，但与对照组相比差异不显著（P>0.05）。由此可见，siRNA TRPV6质粒抑制十二指肠和空肠TRPV6和CaBP-D28K的表达，但不影响血液中钙离子浓度和骨密度水平，表明在正常日粮钙水平条件，沉默TRPV6通道蛋白不影响蛋鸡体内正常的钙转运。

Abstract:

This study was conducted to investigate the effect of transient receptor potential vanilloid receptor 6 (TRPV6) gene silencing on expression of proteins involved in calcium ions transcellular transport in duodenum and jejunum, concentration of calcium and phosphorus and PTH level in plasma, and bone mineral density of femur and tibia in laying hens. Sixty-four laying hens at the peak stage (220 days old) were assigned to control and TRPV6 gene silencing groups, and injected, respectively with physiological saline or interference vector pSIREN-TRPV6-3 in one time, and continuously observed for 28 d. The results were as follows:compared with control group, the expression of ZsGreen protein as an indicator of transfection in duodenum and jejunum were successfully detected at the 7th, 14th and 21st day after injection. The mRNA and protein expression of TRPV6 and CaBP-D28K in duodenum and jejunum exposed to TRPV6 gene silencing were observably decreased at the 7th, 14th and 21st day (P<0.05 or P<0.01). Additionally, the level of PTH in plasma was markedly increased compared with control (P<0.05 or P<0.01). However, the concentration of calcium and phosphorus in plasma were not affected by injection of interference vector pSIREN-TRPV6-3, and the bone mineral density of femur and tibia in the group of TRPV6 gene silencing were slightly lower than control group (P>0.05). It was concluded that the transfection of pSIREN-TRPV6-3 plasmid could suppress the expression of TRPV6 and CaBP-D28K in duodenum and jejunum, but doesn't regulate the concentration of calcium in plasma and bone mineral density, it is implied that silence of TRPV6 gene does not affect normal calcium transport of laying hens under the condition of normal calcium content.

中图分类号:

S852.21

孟范勇, 邓益锋, 杨俊花, 崔军, 周振雷, 侯加法. TRPV6基因沉默对蛋鸡十二指肠和空肠钙离子跨细胞转运相关蛋白表达的影响[J]. 畜牧兽医学报, 2018, 49(11): 2505-2513.

MENG Fan-yong, DENG Yi-feng, YANG Jun-hua, CUI Jun, ZHOU Zhen-lei, HOU Jia-fa. Effects of TRPV6 Gene Silencing on the Expression of Proteins Involved in Calcium Ions Transcellular Transport in Duodenum and Jejunum of Laying Hens[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(11): 2505-2513.

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TRPV6基因沉默对蛋鸡十二指肠和空肠钙离子跨细胞转运相关蛋白表达的影响

Effects of TRPV6 Gene Silencing on the Expression of Proteins Involved in Calcium Ions Transcellular Transport in Duodenum and Jejunum of Laying Hens

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