ITPR1基因过表达对鸭子宫上皮细胞Ca2+浓度、脂质含量及钙转运相关基因的调控效应

Effects of the ITPR1 gene overexpression on Ca2+ concentration, lipid content and calcium transport-related genes in duck uterine epithelial cells

摘要:

肌醇1,4,5-三磷酸受体Ⅰ型 (Inositol 1,4,5-trisphosphate receptor 1，ITPR1) 是细胞内Ca2+释放的重要通道。为探讨ITPR1基因过表达对鸭子宫上皮细胞Ca2+浓度和脂质含量的调控效应及其对钙转运相关基因的影响，文中构建鸭ITPR1基因结构域的真核表达载体，转染鸭子宫上皮细胞，检测细胞ITPR1基因的过表达量、Ca2+浓度、细胞的脂质含量以及其他6个钙转运相关基因的表达量。结果显示，转染后子宫上皮细胞内的Ca2+浓度显著降低 (P<0.05)，甘油三酯含量极显著升高 (P<0.01)，高密度脂蛋白含量极显著降低 (P<0.01)；相关性分析结果显示，C端ITPR1基因过表达量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)，N端ITPR1基因过表达量与甘油三酯含量呈极显著正相关 (P<0.01)，与Ca2+浓度呈显著负相关 (P<0.05)；RT-qPCR结果显示，C端ITPR1基因过表达对IP3R2、VDAC2、CAV1基因的表达有显著抑制作用，N端ITPR1基因过表达对IP3R3、CACNA2D1基因的表达有显著促进作用。总之，ITPR1基因过表达能促进鸭子宫上皮细胞内Ca2+释放，促进甘油三酯、低密度脂蛋白和胆固醇的合成，抑制高密度脂蛋白的生成，且ITPR1基因过表达对6个钙转运相关基因的表达均产生影响。

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.