| Objective:Kidney stones are a non-organic non-fatal urological disease that is common in our lives,accounting for 12%of the global population and this rate is still increasing,posing a significant burden on human health.Given its alarming recurrence rate,it makes it the leading cause of urological hospitalization.The formation of kidney stones is associated with 3 main groups of risk factors:metabolic disorders,defects in renal acidification and urinary tract infections.There is still no single pathogenesis that can correctly explain the pathogenesis of kidney stones.Common theories of kidney stone formation include the Randall’s plaque theory and the Randall’s peg theory,but these theories of kidney stone formation have not been clearly elaborated to date.Immune cells are widely involved in the regulation of the immune function of the body,and it has been shown that immunological alterations are thought to be closely related to the formation of Randall’s plaque,but their specific effects are unknown to us.Therefore,clarifying the relationship between immune cells and kidney stone formation may be key to the future treatment of kidney stones.In this study,we explored the role of immune cells in the formation of kidney stones to provide new ideas for the future treatment and prevention of kidney stones.Methods and Materials:1.Construction of SD rat kidney stone modelTwenty-eight SD rats aged 6-8 weeks were randomly divided into two groups,the stone group and the control group,with 14 rats in each group.The calcium oxalate kidney stone model was induced by freely drinking ethylene glycol solution and ammonium chloride solution in the stone group,while the control group was freely drinking distilled water and ammonium chloride solution.24-h urine was collected from SD rats after 28 d of modeling,and the kidney tissues were extracted by executing the rats.2.Verification of kidney stone formation in SD rat kidney stone model.The formation of urinary crystals was observed by microscopy,and the formation of calcium crystals and inflammatory infiltration around the kidney tissue in histopathological sections of the kidneys of SD rats in the kidney stone and control groups were detected by CT,Alizarin Red S staining,Von Kossa staining,Pizzolatto’s staining and HE staining of the kidneys.3.Gene expression profiles of kidney tissues of SD rats were analyzed by RNA-seq.RNA was extracted from the kidney tissues of the five kidney stone groups and the kidney tissues of the five control groups with significant differences,and RNAseq was done to observe the gene expression in the stone and control groups and to compare the gene expression differences between the kidney stone and control groups.4.Observe the expression of immune cells in kidney tissues of SD rats by multiple immunofluorescence staining.Paraffin sections of kidney tissues from 5 groups of kidney stone group and 5 groups of control group with significant differences were taken for multicolor immunofluorescence staining to observe the expression of relevant immune cells in the tissues of kidney stone group and control group,and the expression differences were compared.Results:1.By observing the results of urine crystals,kidney CT scans,Alizarin Red S staining,Pizzolatto’s staining,Von Kossa staining and H&E staining,we proved that the kidney stone model in SD rats was successfully constructed and confirmed the presence of inflammatory infiltration in the kidney tissue around the kidney stones.2.The RNA-seq results showed that there were significant inflammatory changes in the kidney tissues of SD rats in the kidney stone group,with more inflammatory factors and activation of inflammatory pathways compared to the kidney tissues of SD rats in the control group.3.Multicolor immunofluorescence results showed increased infiltration of CD45+immune cells,CD3+T cells,CD4+T cells,and F4/80+macrophages in the kidney tissues of SD rats in the kidney stone group compared to the kidney tissues of SD rats in the control group.Conclusions:In this paper,we used to construct SD rat calcium oxalate kidney stone model,and by analyzing the kidney tissue of SD rats,the results showed that immune remodeling had an important role in kidney stone formation in SD rats,in which CD45+immune cells,CD3+T cells,CD4+T cells and F4/80+macrophages were the key cells of immune remodeling in the kidney stone model of SD rats. |
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