| [Objective] To compare the differences and significance of rat kidney stone model established by nanobacteria(NB)and rat kidney stone model established by ethylene glycol(EG).[Methods] A total of 90 Wistar male rats were randomly divided into nanobacteria group(NB group),ethylene glycol group(EG group),and control group(NC group).At the end of the first week of modeling,three rats were randomly selected from each group once a week for ten times.Blood and urine metabolism were measured.After sacrifice,stone formation was observed.The stone composition was analyzed by infrared spectroscopy.Ultrastructural changes of the kidney were observed by electron microscopy and pathology.[Results] Compared with NB group,EG group had less mental fatigue,activity and food intake.The kidney to body weight ratio of rats in the NB group began to be higher t han that of the NC group at the 8th week,while that of the EG group began to be higher than that of the NC group at the 3rd week;the biochemical metabolic parameters of rats in the NB group increased at the 3rd week and would reach to normal at the 9th week,while those of rats in the EG group increased at the 2nd week and did not return to normal at the later stage,and the serum creatinine level of rats in the EG group was significantly higher than that of rats in the NB group at the 7th week.The difference was statistically significant.The stone formation rate in NB group and EG group was 52.4% and 66.7%,respectively,which was slightly lower in NB group,but there was no significant difference in stone formation rate between the two groups.[Conclusion] Both methods were able to form stones.There was no statistically significant difference in stone formation rate between the two methods.In NB group,stone formation was slightly later.However,NB model had less damage to rat kidney,and the process of stone formation was more consistent with the natural course of kidney stones.It was more meaningful to study the etiology of kidney stones.[Objective] To study the role of nanobacteria in the formation of renal calculi.[Methods] A total of 90 clean Wistar male rats were randomly divided into negative control group,experimental group and interference group.From the end of the first week of modeling,once a week for ten consecutive times,three rats in each group were randomly selected to detect the blood and urine metabolism.After sacrifice,the formation of stones was observed.The ultrastructure of the kidney was observed by electron microscopy and pathology.[Results] Compared with the control group,the gross structure of the kidney was changed: At the 4th week after modeling,the rats in the nanobacteria group had significantly enlarged kidneys and increased kidney-to-body ratio,and the difference had statistical significance(P<0.05).The color of the kidney profile was dark,the structure of the skin pulp was less clear,and the accumulation of yellowish particles was observed at the junction of the skin pulp.The metabolism of blood and urine: the creatinine,uric acid,urea nitrogen and urinary calcium of the rats in the nanobacteria group began to increase at the 3rd week,and the difference between the 3rd and 8th week had statistical significance(P<0.05);after the 8th week,the difference between the three groups had no statistical significance.At the 4th week,there was calculus formation,which was mainly distributed in the renal tubules and surrounding tissues.The stone formation rate was 52.4% in nanobacteria group and 27.8% in interference group,and the difference had statistical significance(P < 0.05).Ultrastructure: From the 4th week,the renal tissues in nanobacteria group showed expanded renal tubules,swollen renal tubular epithelium,granular degeneration,shedding and lymphocyte infiltration of renal tubular epithelial cells,and a small amount of calcium salt crystals in renal tubules.[Conclusion] 1.The formation of renal calculi began in the 4th week after the model was established,and the crystals were mostly located in the renal tubules.2.During the formation of renal calculi,the renal tubular epithelial cells were damaged,as follows: granular degeneration of renal tubular epithelial cells,a small amount of calcium salt crystal accumulation in the renal tubules,accompanied by a few renal tubules began to expand and epithelial swelling,granular degeneration,necrosis and shedding of renal tubular epithelial cells,lymphocyte infiltration in the renal interstitium,a small amount of calcium salt crystal deposition in the renal tubules,aggravated with time;3.The serum creatinine,serum uric acid,urea nitrogen and urinary calcium levels increased with time from the 3rd week,and returned to normal after the 8th week.[Objective] To study the expression of Ca SR and claudin-14 proteins in kidney tissues of experimental rats induced by nanobacteria.[Methods] A total of 120 clean wistar male rats were randomly divided into negative control group(NC group,normal saline+normal saline gavage),experimental group(NBS group,NB+normal saline gavage),interference group(NBT group,NB+tetracycline gavage)and positive control group(EG group,ethylene glycol gavage).After the modeling,the rats were sacrificed and their kidneys were extracted.The expressions of Ca SR and claudin-14 in renal tissues were detected by immunohistochemistry,q RT-PCR and Western blotting,respectively.[Results] Compared with the NC group,Ca SR and claudin-14 in each group were expressed more strongly at 3rd week in NBS,NBT and EG groups,and the expression became more obvious with the extension of time.Ca SR was consistently expressed in the NC group,while claudin-14 was not significantly expressed in the NC group.Immunohistochemistry,qrt-PCR and Western blot showed: The expressions of Ca SR and claudin-14 in NBS group were significantly higher than those in NBT group and NC group,with statistically significant differences(P<0.05).The expressions of Ca SR and claudin-14 in EG group were significantly higher than those in NBS group,NBT group and NC group,with statistically significant differences(P <0.05).[Conclusion] From the 3rd week,the protein expression of Ca SR and claudin-14 in each group was up-regulated,which became more obvious with the extension of action time.Ca SR and claudin-14 expression in NBT group was lower than that in EG group,which may be related to the fact that the nanobacterium had less renal d amage than glycol.[Objective] This study was designed to investigate the mechanism of nanobacteria in the formation of kidney stones by detecting the injury of renal tubular epithelial cells and the adhesion to crystals.[Methods] Renal tubular epithelial cells were selected and randomly divided into 4 groups: negative control group(Control group),nanobacteria group(NB group),interference group(TET + NB group),and positive control group(COM group).The cells were cultured in the same environment.The injury indicators of renal tubular epithelial cells were collected in different time periods.The cells were collected.The morphological changes of renal tubular epithelial cells were observed by transmission electron microscopy.The adhesion of cells to crystals was observed by confocal laser scanning microscopy.[Results] Compared with the control group,HE staining results: In the NB group,at 6 h after the start of the experiment,some cells showed increased cell body expansion,loose nuclei and indistinct nuclear membrane,and the injury was gradually aggravated with the extension of the action time.Transmission electron microscopy results: in the NB group,the microvillus structure was slightly disordered,the local villus disappeared,the nucleus was deformed,the chromatin was slightly unevenly distributed,the local area of the nuclear outer membrane was degraded,some mitochondrial structures were slightly swollen,and myeloid bodies were occasionally found in the cytoplasm.Damage factor index test results: after 12 h and 24 h of co-culture,the Na+-K+-ATPase and Ca2+-Mg2+-ATPase activities of the NB group and COM cells were lower than those of the control group(P < 0.05).The H2O2 and MDA contents in the culture medium of NB group and COM group were significantly higher than those of the control group(P < 0.05).Adhesion results: at 6 h of co-culture,the adhesion amount in the NB group was less than that in the COM group.At 12 h and 24 h,the adhesion amount of crystals in the NB group was further increased than that before.The difference was statistically significa nt(P < 0.001)[Conclusion] 1 Nanobacteria can damage HK-2 cells through lipid peroxidation reaction,and the adhesion to crystals is increased after cell injury.The amount of crystal adhesion is proportional to the time of nanobacteria action.Anti-nanobacteria can protect HK-2 cells and reduce their adhesion to crystals.2 During the damage of HK-2 cells by nanobacteria,the activity of Ca2+-Mg2+-ATPase is decreased,the transport of Ca2+ is blocked,the concentration of extracellular Ca2+ is increased. |
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