Department of Nutrition Science, Purdue University, West Lafayette, IN 47907-2059, USA.
J Bone Miner Res. 2012 Oct;27(10):2097-107. doi: 10.1002/jbmr.1662.
Transient receptor potential cation channel, subfamily V, member 6 (TRPV6) is an apical membrane calcium (Ca) channel in the small intestine proposed to be essential for vitamin D-regulated intestinal Ca absorption. Recent studies have challenged the proposed role for TRPV6 in Ca absorption. We directly tested intestinal TRPV6 function in Ca and bone metabolism in wild-type (WT) and vitamin D receptor knockout (VDRKO) mice. TRPV6 transgenic mice (TG) were made with intestinal epithelium-specific expression of a 3X Flag-tagged human TRPV6 protein. TG and VDRKO mice were crossed to make TG-VDRKO mice. Ca and bone metabolism was examined in WT, TG, VDRKO, and TG-VDRKO mice. TG mice developed hypercalcemia and soft tissue calcification on a chow diet. In TG mice fed a 0.25% Ca diet, Ca absorption was more than three-fold higher and femur bone mineral density (BMD) was 26% higher than WT. Renal 1α hydroxylase (CYP27B1) mRNA and intestinal expression of the natural mouse TRPV6 gene were reduced to <10% of WT but small intestine calbindin-D(9k) expression was elevated >15 times in TG mice. TG-VDRKO mice had high Ca absorption that prevented the low serum Ca, high renal CYP27B1 mRNA, low BMD, and abnormal bone microarchitecture seen in VDRKO mice. In addition, small intestinal calbindin D(9K) mRNA and protein levels were elevated in TG-VDRKO. Transgenic TRPV6 expression in intestine is sufficient to increase Ca absorption and bone density, even in VDRKO mice. VDR-independent upregulation of intestinal calbindin D(9k) in TG-VDRKO suggests this protein may buffer intracellular Ca during Ca absorption. © 2012 American Society for Bone and Mineral Research.
瞬时受体电位阳离子通道,亚家族 V,成员 6(TRPV6)是小肠中的一种顶端膜钙(Ca)通道,被认为是维生素 D 调节肠道 Ca 吸收所必需的。最近的研究对 TRPV6 在 Ca 吸收中的作用提出了质疑。我们直接在野生型(WT)和维生素 D 受体敲除(VDRKO)小鼠中测试了肠道 TRPV6 在 Ca 和骨代谢中的功能。TRPV6 转基因(TG)小鼠通过肠上皮细胞特异性表达 3X Flag 标记的人 TRPV6 蛋白制成。TG 和 VDRKO 小鼠被杂交以制造 TG-VDRKO 小鼠。在 WT、TG、VDRKO 和 TG-VDRKO 小鼠中检查 Ca 和骨代谢。TG 小鼠在普通饮食中出现高钙血症和软组织钙化。在 TG 小鼠喂食 0.25%Ca 饮食时,Ca 吸收增加了三倍以上,股骨骨密度(BMD)比 WT 高 26%。肾 1α羟化酶(CYP27B1)mRNA 和肠道天然小鼠 TRPV6 基因的表达降低至 WT 的<10%,但 TG 小鼠的小肠钙结合蛋白-D(9k)表达升高了 15 倍以上。TG-VDRKO 小鼠的 Ca 吸收很高,可防止 VDRKO 小鼠出现低血清 Ca、高肾 CYP27B1 mRNA、低 BMD 和异常骨微结构。此外,TG-VDRKO 中小肠钙结合蛋白-D(9K)mRNA 和蛋白水平升高。肠内 TRPV6 的转基因表达足以增加 Ca 吸收和骨密度,即使在 VDRKO 小鼠中也是如此。在 TG-VDRKO 中,VDR 不依赖的肠钙结合蛋白-D(9k)上调表明该蛋白在 Ca 吸收期间可能缓冲细胞内 Ca。©2012 美国骨矿研究协会。