摘要
为了探讨瞬时性受体电位通道香草酸受体5、6(TRPV5、TRPV6)在胎盘滋养层(CT)细胞钙转运中的作用。取孕7.5 d昆明小鼠胎盘组织分离纯化并鉴定CT细胞,重组慢病毒颗粒TRPV5 siRNA/shRNA/RNAi Lentivirus、TRPV6 siRNA/shRNA/RNAi Lentivirus感染CT细胞,实时荧光定量RT-PCR、Western Blot分别检测转染后细胞内TRPV5/6 mRNA及蛋白表达量,激光共聚焦扫描显微镜经钙离子成像分析细胞内Ca^(2+)浓度。结果显示:与无慢病毒载体感染的空白对照组比较,TRPV5/6 siRNA/shRNA/RNAi Lentivirus感染CT细胞后,TRPV5/6 mRNA及蛋白表达水平均下调(P<0.05),细胞内钙水平显著性降低(P<0.05),慢病毒转染的2个组间的细胞钙荧光强度未见显著差异(P>0.05)。结果表明,TRPV5和TRPV6参与了CT细胞的钙转运,对妊娠期胎盘钙转运具有重要的促进作用。
The Ca^(2+) transport effect of the calcium channel transient receptor potential vanilloid 5 and 6(TRPV5 and TRPV6)in placental cytotrophoblasts was investigated.The placental cytotrophoblast cells from pregnant day 7.5 mice were isolated and purified.Then,the cytotrophoblasts were transfected by lentivirus particles of TRPV5 siRNA/shRNA/RNAi Lentivirus and TRPV6 siRNA/shRNA/RNAi Lentivirus,respectively.Real-time quantitative reverse transcription PCR(qRT-PCR)and Western Blot were used to detect the expression of TRPV5/6 mRNA and proteins,respectively.Cellular calcium was determined by Rhod-2-AM calcium imaging using confocal laser scanning microscope(CLSM).The results revealed the significant down-regulation of both TRPV5 and TRPV6 mRNA and protein expression and intracellular calcium contents in comparison with the non-transfected cytotrophoblasts in the control groups(P<0.05).Thus,TRPV5 and TRPV6 may mediate the calcium transport of placental cytotrophoblast and may play a role in maternal-fetal calcium metabolism during pregnancy.
作者
季佳
黄小兰
尹红斌
李素华
黄景军
栾云鹏
JI Jia;HUANG Xiao-lan;YIN Hong-bin;LI Su-hua;HUANG Jing-jun;LUAN Yun-peng(College of Life Sciences,Southwest Forestry University,Kunming 650024,China;Animal Disease Inspection and Supervision Institution of Yunnan Province,Kunming 650051,China)
出处
《中国兽医杂志》
CAS
北大核心
2021年第8期42-47,共6页
Chinese Journal of Veterinary Medicine
基金
国家自然科学基金(31260630)。
