Efficacy Of New Bisphosphonate To Inhibit Vascular Calcification In5/6Nephrectomized Rats

The world each year about10%-13%of people suffering from chronic kidney disease,cardiovascular disease is the leading cause of death in chronic kidney disease, end stagerenal failure complicated by cardiovascular disease mortality is age-matched generalpopulation of10-20times. End-stage renal failure complicated byhyperphosphatemia-mediated vascular calcification, and the resulting hardening of thearteries is an important cause of cardiovascular disease complications of chronic renalfailure, and the high incidence of cardiovascular disease in chronic kidney disease, highcase fatality rate is close related. Hyprphosphatemia-mediated mechanism of vascularcalcification in chronic kidney disease patients is not very clear, the clinical treatment ofchronic kidney disease patients with vascular calcification in drugs is extremely limited, tofind an effective therapy has become the clinical problems to be solved. Therefore, fromcell and animal experiments on six kinds of new bisphosphonates inhibit the efficacy ofvascular calcification study. In this study, divided into four parts: a rat aortic vascularsmooth muscle cells in vitro in primary and subcultured; two new bisphosphonateintervention in the high phosphorus cultured rat vascular smooth muscle cell calcificationin cell experiments screening;5/6nephrectomized rats heightening phosphorus dietinduced vascular calcification in animal models of chronic renal failure; efficacy of four, anew bisphosphonate to inhibit vascular calcification in5/6nephrectomized rats.一、Rat vascular smooth muscle cells in vitro in primary and subculturedRat vascular smooth muscle cells access to convenient sources of economic, suitablefor cultivation and experimental. Vascular smooth muscle cells in vitro and passage culture,the main enzyme digestion, tissue explants and adherent method, different trainingmethods have their own complexity, advantages, disadvantages and usefulness. Theenzyme digestion method cell digestive enzyme digestion time is difficult to control,expensive digestive enzymes, the cells acquire sometimes enough, but the availability ofwell-differentiated cell morphology and contractile phenotype cells; explants adherentmethod easily obtained, although the primary cells of can the shortcomings of passagedcells in a long time, the low success rate, vascular smooth muscle cells were subculturedreceive a large number of homogeneous cells, and can maintain the basic characteristics ofthe vascular smooth muscle cells in vitro, very suitable for the cells require a large amount of experimental. We use the explant adherent successfully cultured in vitro rat aorticvascular smooth muscle cells, vascular calcification is a good and practical cell material.二、New bisphosphonate intervention in the high phosphorus cultured rat aorticvascular smooth muscle cell calcification in cell experiments screeningWe choose to grow well in the3rd generation rat vascular smooth muscle cells, adjustcell density inoculated in6-well plates. Cell fusion induced by75-80%by adding inducedby high phosphorus (Pi2.5mmol/L), the calcification of vascular smooth muscle cellscultured for3days and at the same time points the concentration gradient of abisphosphonate intervention, intervention after cresol phthalate complexation ketoneQuantitative Determination of calcium content in the extracellular matrix, and rat aorticvascular smooth muscle cells of qualitative research von Kossa staining, observed a newtype of bisphosphonate calcification inhibition of rat aortic vascular smooth muscle cellscultured in high phosphorus and classic pairs bisphosphonates pamidronate compared withthe new bisphosphonate filter, screening out the new bisphosphonate further animalexperiments, bisphosphonates inhibit the high phosphorus cultured vascular smooth musclecell calcification mechanism to conduct a preliminary Discussion.三、5/6nephrectomized rats to heightening phosphorus diet for vascular calcificationcaused by chronic renal failure in animal modelsAccording to the etiology, pathogenesis and prevention of different studies, thepathogenesis of vascular calcification caused by the different models to clarify the differentcauses and prevention and treatment of vascular calcification and development of newdrugs is of great significance. Preparation of vascular calcification in animal models ofmethods such as drug-induced knockout inbred training of high-fat and high cholesterolfeeding, although the preparation process have their own complexity, but the pathogenicmechanism perspective does not apply to simulated chronic renal failure, calcium andphosphorus metabolism disorders caused by vascular calcification. Therefore, we followthe pathogenesis of ideas on the basis of SD rats one-step5/6kidney removal and at thesame time be the high-phosphorus diet simulate chronic renal failure in patients withcalcium and phosphorus metabolism disorders due to vascular calcification.28days afterthe surgery group, serum creatinine, blood urea nitrogen was significantly highersymptoms of hypocalcemia, hyperphosphatemia in chronic renal failure, and confirmed bypathological observation5/6nephrectomy rats with chronic renal failure models, modeling, through von Kossa staining, rat aortic vascular pathology observed the presence ofvascular calcification, and to lay the foundation for animal experiments.四、Efficacy of new bisphosphonates to inhibit vascular calcification in5/6nephrectomized rats.In drug experiments, in vitro and in vivo experiments have their own advantages anddisadvantages: in vitro experiments under the conditions of man-made environment andthe body a lot of difference, the advantage of being simple, fast, economic, and use a verysmall amount of drugs a short time a large number of experimental data, the drawback isthe false positive rate; in vivo screening experiments the drugs in the conditions of theenvironment as a whole, the advantage of the credibility of its results, predictability ofclinical efficacy, under normal circumstances. The in vivo experimental results, is alsoeffective in clinical practice. The in vivo experimental results and clinical observationsrelated to a higher rate of false positive rate and false negative rates are low, its drawbackis that the experimental procedures, long cycle, the high cost and large amount ofmedication.Cell experiments in the previous study, we screened a new bisphosphonate, ofbisphosphonates to inhibit the high phosphorus cultured rat aortic smooth muscle cellcalcification mechanism, and successfully established chronic renal failure, calciumphosphorus metabolism disorders vascular calcification in animal models. In this section,we will observe the new bisphosphonates in animals, the inhibition of vascularcalcification in5/6nephrectomized rats, as well as salts with classical diphosphinepamidronate calcification inhibitory effect of the comparison, and new bisphosphonates inthe reversal of vascular calcification in a preliminary study.In summary, through the cells of the new bisphosphonate experimental screening andanimal experiments efficacy observations confirm the new bisphosphonate to inhibit the5/6nephrectomy, the efficacy of vascular calcification is better than the classicbisphosphonate Pamidronate. Further research will help clarify the bisphosphonates inhibitthe mechanisms of vascular calcification, and clinically effective treatment of chronic renalfailure, calcium and phosphorus metabolism disorders mediated by the vascularcalcification provides a new option, provide a reference for the development of new drugs.