Abstract:
Inositol 1,4,5-trisphosphate receptor 1 (ITPR1) is an important intracellular channel for releasing Ca²⁺. In order to investigate the effects of the ITPR1 overexpression on Ca²⁺ concentration and lipid content in duck uterine epithelial cells and its effects on calcium transport-related genes, the structural domain of ITPR1 gene of duck was cloned into an eukaryotic expression vector and transfected into duck uterine epithelial cells. The overexpression of the ITPR1 gene, the concentration of Ca²⁺, the lipid content, and the expression of other 6 calcium transport-related genes was determined. The results showed that the concentration of Ca²⁺ in uterine epithelial cells was significantly reduced after transfection (P<0.05), the triglyceride content was significantly increased (P<0.01), and the high-density lipoprotein content was significantly decreased (P<0.01). The correlation analysis results showed that the overexpression of the C-terminal half of the ITPR1 gene was significantly positively correlated with the total cholesterol content (P<0.01), which was significantly positively correlated with the low-density lipoprotein content (P<0.05). The overexpression of the N-terminal half of the ITPR1 gene was significantly positively correlated with the triglyceride content (P<0.01), which was significantly negatively correlated with the concentration of Ca²⁺ (P<0.05). RT-qPCR results of 6 calcium transport-related genes showed that the overexpression of the C-terminal half of the ITPR1 gene significantly inhibited the expression of the IP3R2, VDAC2 and CAV1 genes, and the overexpression of the N-terminal half of the ITPR1 gene significantly promoted the expression of the IP3R3 and CACNA2D1 genes. In conclusion, the ITPR1 gene overexpression can promote Ca²⁺ release in duck uterus epithelial cells, promote the synthesis of triglyceride, low-density lipoprotein and cholesterol, and inhibit the production of high-density lipoprotein, and the ITPR1 gene overexpression affected the expression of all 6 calcium transport-related genes.
摘要:
肌醇1,4,5-三磷酸受体1(ITPR1)是释放Ca²的重要细胞内通道。为了探讨ITPR1过表达对鸭子宫上皮细胞Ca²+浓度和脂质含量的影响及其对钙转运相关基因的影响,将鸭ITPR1基因的结构域克隆到真核表达载体中,并转染到鸭子宫上皮细胞中。ITPR1基因的过表达,Ca²的浓度,脂质含量,并测定了其他6个钙转运相关基因的表达。结果表明,转染后子宫上皮细胞中Ca²的浓度明显降低(P<0.05),甘油三酯含量显著升高(P<0.01),高密度脂蛋白含量显著降低(P<0.01)。相关性分析结果显示,ITPR1基因C端一半的过表达与总胆固醇含量呈极显著正相关(P<0.01),与低密度脂蛋白含量呈显著正相关(P<0.05)。ITPR1基因N端一半过表达与甘油三酯含量呈显著正相关(P<0.01),与Ca²+浓度呈显著负相关(P<0.05)。6个钙转运相关基因的RT-qPCR结果显示,ITPR1基因C端一半的过表达显著抑制IP3R2、VDAC2和CAV1基因的表达,ITPR1基因N端一半的过表达显著促进IP3R3和CACNA2D1基因的表达。总之,ITPR1基因过表达可促进鸭子宫上皮细胞释放Ca²。促进甘油三酯的合成,低密度脂蛋白和胆固醇,抑制高密度脂蛋白的产生,ITPR1基因过表达影响了所有6个钙转运相关基因的表达。