The Effect Of Bone Resorption Inhibitor Alendronate On Bone Metabolism Of TRPV5^-/- Mice

Gene knockout is a self-developed since the late 80s a new type of molecular biology techniques, is the body through a certain way so that a particular gene inactivation or deletion of technology. Gene knockout technology, commonly used in the establishment of a specific gene deletion in animal models, which carry out related research. By gene knockout to determine the nature of specific genes and to study the impact of its body, whether it is understanding the root causes of the disease or to find the target goal of gene therapy are of great significance. At present, the gene knockout mice to study the occurrence of the disease mechanism, diagnosis and treatment of the molecular basis of important experimental material. TRPV5 is a calcium ion-selective channels involved in calcium absorption and re-absorption, its main expression in the kidney distal convoluted/collecting duct, duodenum, and bone osteoclasts fold edge. Alendronate is a third generation of bone resorption inhibitor, at the cellular level, alendronate, primarily by reducing the number of osteoclasts, inhibit osteoclast formation of edge folds, lower mature osteoclast lysosomal enzyme the release, which will reduce osteoclast function. In this study, the introduction of TRPV5 heterozygous knock-out mice, feeding and breeding in accordance with the requirements SPF-class standards of animal husbandry carried out three kinds of offspring phenotype may occur, namely, the wild-type TRPV5+/+, heterozygous TRPV5+/- and homozygous TRPV5-/-, thus a need to conduct genotyping of the offspring, thereby screening out the wild-type TRPV5+/+ and homozygous TRPV5-/-. In order to study of alendronate on TRPV5+/+ and TRPV5-/- mice with bone, kidney, duodenum calcium ion channel expression, this experiment will TRPV5+/+ and TRPV5-/- mice were divided into four groups kept in metabolic cages, in which two groups of mice of different genotypes were given alendronate weekly 2mg/kg once a row for 8 weeks, collecting blood and urine specimens and do biochemical analysis, mouse femur, kidney, duodenum sample preparation. Extracted from mouse femur bone broken cell culture, and its surface folds calcium channel on the edge of a quantitative polymerase chain reaction analysis, application uCT right femur scan, and calcium absorption peak for testing. Application of alendronate treated TRPV5+/+ mice increased the thickness of the femur, while the TRPV5-/- mouse femoral thickness reduced to normal, the expression of both TRPV5 was significantly increased, while the kidney and the duodenum calcium channel has not changed. Two kinds of genotypes of mice osteoclasts folds on the edge of CLC-7 protein and vacuolar H+-ATPase expression in no significant change, however, alendronate significantly increased TRPV5-/- mice, PTH hormone levels and expression. Osteoclast culture medium TRPV5, CLC-7, and the vacuolar H+-ATPase were not affected by the effects of alendronate, TRPV5-/- mice with bone marrow culture to reduce bone resorption, the two pairs of vitamin D, Reaction was no significant difference. Alendronate prompted TRPV5+/+ mice TRPV5 genes increases, which will reduce TRPV5-/- mice osteoclasts absorption shows TRPV5 functions in osteoclasts play a very important role. Studies have shown that TRPV5-/- mice, bone re-absorption process is still ongoing, because alendronate can TRPV5-/- mice, normalization of bone thickness.