动物造模,药效评价,高血压肾阴虚证 z-bio 2024-07-29 473

　　(1) Adult rats were replicated by removing one kidney and ligating 2/3 of the renal artery on the other side. Blood pressure in the tail artery of the rats was measured 2 weeks after surgery. Rats with systolic blood pressure>140mmHg (18.6kPa) were identified as hypertensive rats. Animals were euthanized at 4 time points: 2 weeks, 1 month, 2 months, and 4 months after surgery. Before euthanization, 24-hour urine samples were collected and 24-hour urine protein levels were measured; Then take blood from the heart and measure the concentration of creatinine and urea nitrogen; Execute the animal and immediately remove the kidney, prepare histopathological samples and observe relevant indicators.

　　(2) Model characteristics: Blood pressure of experimental animals increased at 2 weeks, 1 month, 2 months, and 4 months after surgery. At 2 and 4 months after surgery, blood creatinine, urea nitrogen concentrations, and 24-hour urine protein levels significantly increased, indicating severe damage to renal function. One month after surgery, the glomerular basement membrane (GBM) of animals showed slight thickening, focal mesangial cell proliferation, and increased mesangial matrix. Two months after the operation, the mesangial matrix increased significantly, especially the blood vessels. The deposition of PAS positive substances and foam like changes were seen. The number of mesangial cells increased significantly, and crescent formation was seen in some cases. At 4 months, there was an increase in the number of hardened glomeruli, with strong PAS positive glass like material deposition along the vascular wall, and significant thickening of GBM. Under ordinary electron microscopy, it can be seen that the mesangial area of animal glomeruli widens and tends to increase in matrix after 2 weeks of surgery. At 1 month after surgery, the mesangial matrix increases, the number of cells increases, and there is edema under the endothelial cells. The thickness of the basement membrane varies. At 2 months of the disease, the basement membrane generally thickens. At 4 months after surgery, the endothelial cells become edematous and degenerate, the endothelial cell pores open, the mesangial matrix increases significantly and hardens, and there is a proliferation of basement membrane like substances and dense deposits, resulting in significant widening of the mesangial area and thickening of the basement membrane. Small hill like protrusions can be formed towards the epithelial side, and foot process fusion can be observed. Two and four months after surgery, the GBM of hypertensive rats showed significant thickening. Histochemical electron microscopy reveals that the outer loose layer of GBM in normal animals contains PEI stained particles of uniform size and regular arrangement (this is the location where negative charge sites exist). There is a significant increase in PEI stained particles at the inflection point of the basement membrane, and there are also a small number of PEI stained particles with lighter staining, inconsistent size, and irregular arrangement in the loose layer of the basement membrane. PEI stained particles on GBM have a large quantity and high density. Scattered PEI stained particles can also be seen in the glomerular mesangial matrix. As the disease progresses, PEI stained particles gradually decrease in model animals. Four months after surgery, the number of PEI stained particles in the loose outer layer of GBM in the model animals decreased significantly, with sparse arrangement and irregular arrangement caused by particle loss. Polyimine (PEI) stained particles in the glomerular mesangial matrix also gradually decrease with the progression of the disease.

    (3) The theoretical basis of the comparative medicine model is mainly based on the fact that hypertension is generally characterized by a syndrome of kidney yin deficiency and internal heat. This model uses nourishing kidney yin formula to be effective, while using warming kidney yang formula can actually worsen the condition. The traditional Chinese medicine syndrome differentiation for clinical hypertension is mostly liver kidney yin deficiency syndrome or liver yang hyperactivity syndrome. This method simulates the pathogenesis of hypertension and replicates the hypertension disease model. However, whether the syndrome characteristics belong to simple kidney yin deficiency syndrome needs further evaluation, and it is difficult to determine based solely on the above experimental results.