【Animal Modeling】-Urinary Tract Stone Rat Model

  [Step] The male Wistar rats weighed 180-220 g, were randomly divided into several groups, and fed with normal diet. Before the end of the experiment, a metabolic cage was used to collect and measure the 24-hour urine output and 24-hour oxalic acid (Ox). ) Fecal matter (hydrogen peroxide-hydrochloric acid). Phenylhydrazine method). Give 1% ethylene glycol (EG) drinking water and 0.5μgla-(OH)-VitD3 to the stomach every other day for 28 days. The killed rats were anesthetized and blood was collected by inferior vena cava puncture. After each heart was centrifuged at a speed of 3500 pm for 10 minutes, the upper serum was collected to detect the contents of BUN, Cr, P and Ca2 +. Cut both kidneys and cut vertically. The left kidney was taken and fixed with 10% neutral formalin. Perform conventional HE paraffin sections and von Kossa staining. Observe the morphological distribution of calcium oxalate crystals in the kidney tissue and the pathological changes of the kidney tissue. Absorption spectrometer is used to measure the content of Ca2 + and Mg2 + in kidney tissue and the total content of Ca2 + and Mg2 + in urine.

  [Result analysis] Most people's urinary stones are calcium-containing stones, and most of them are deposited in the urethra in the form of calcium oxalate. Provide animals with active vitamin D3 to increase the absorption of calcium ions in the intestine; provide ethylene glycol to increase the production of oxalic acid in the body and promote the excretion of oxalic acid in the urine. This causes calcium oxalate to deposit in the urethra and form stones. .. 7 days after the experiment, the excretion of calcium oxalate began to increase significantly, and the excretion of urinary phosphorus and urinary magnesium also began to increase significantly. After 14 days, the blood calcium level tended to rise significantly, and the blood phosphorus also tended to rise, but the blood magnesium did not change significantly. Blood urea nitrogen began to increase after 2 weeks and increased significantly after 3 weeks. At this time, the inulin clearance rate also decreased significantly. After 4 weeks, the kidney was significantly enlarged, stones were found in the renal pelvis and calyx, or radial calcium deposits were found in the junction of the renal cortex, bone marrow and papilla. Obvious renal tubule expansion can be seen under an optical microscope, and colorless transparent crystal deposits can be seen in the cavity; oxalic acid yellow crystals can be observed under a polarized light microscope; renal tubules in the distance can be seen by yonKossa staining , There is black calcium deposited in the collection tube lumen.

  This experiment is the result of a comparative study of the rat kidney calcium oxalate stone model induced by single and complex stone attractants commonly used at home and abroad. 1% EG drinking water + 0.5μg 1α-(OH)-VitD3 1% EG drinking water + 2% acetylcysteine (AC) 2 ml per day (4 weeks) intragastric administration (4 weeks) is a rat stone formation The most common method produces important, simple and stable effects.