Yang Heng, Cheng Xiaofeng, Chen Yujun, Zeng Zhenhao, Wang Gongxian
1 Affiliated Hospital, Jiangxi Medical College, Nanchang University, China.
Arch Med Sci Atheroscler Dis. 2024 Jan 30;9:e1-e15. doi: 10.5114/amsad/177534. eCollection 2024.
The aim of the study was to study the role of nanobacteria in the formation of renal calculi and the underlying mechanism.
A total of 90 clean Wistar male rats were randomly divided into a negative control group, an experimental group, and an interference group. From the end of the first week of modelling, 10 consecutive times once a week, 3 rats in each group were randomly selected to measure the biochemical blood markers and urine metabolism. After sacrifice, the formation of kidney stones was assessed by observing the ultrastructure of the kidney by electron microscopy and pathohistology. Finally, the expression of calcium-sensitive receptor (CaSR) and claudin-14 protein in the kidney tissue was examined by western blotting.
Compared with the control group, the gross structure of the kidney was changed in the model group. At the fourth week of modelling, the rats in the nanobacteria group had significantly enlarged kidneys and increased kidney-to-body ratio, and the difference had statistical significance ( < 0.05). The colour 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 cortical pulp. The creatinine, uric acid, urea nitrogen, and urinary calcium of the rats in the nanobacteria group began to increase at the third week, and the difference between the third and eighth week had statistical significance ( < 0.05). However, the difference between the 3 groups had no statistical significance after the eighth week. At the fourth week, we observed the formation of calculi, which were mainly distributed in the renal tubules and surrounding tissues. The kidney stone formation rate was 52.4% in the nanobacteria group and 27.8% in the interference group, and the difference had statistical significance ( < 0.05). Ultrastructure observations revealed that from the fourth week, the renal tissues in the 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. At the third week, the expression of CaSR and Claudin-14 protein in the nanobacteria group increased, and the difference had statistical significance ( < 0.05). The expression of CaSR and Claudin-14 was positively correlated with urinary calcium ( < 0.05).
The formation of renal calculi began in the fourth week after the model was established, and the crystals were mostly located in the renal tubules. During the formation of renal calculi, the renal tubular epithelial cells were damaged, showing granular degeneration and small amounts of calcium salt crystals, accompanied by a few renal tubules beginning to expand and epithelial swelling, granular degeneration, necrosis and shedding of renal tubular epithelial cells, lymphocyte infiltration in the renal interstitium, and small amounts of calcium salt crystals in the renal tubules, which aggravated with time. The serum creatinine, serum uric acid, urea nitrogen, and urinary calcium levels increased with time from the third week and returned to normal after the eighth week. The expression of CaSR and Claudin-14 protein was upregulated and positively correlated with the 24-h urinary calcium excretion value.
本研究旨在探讨纳米细菌在肾结石形成中的作用及其潜在机制。
将90只清洁级Wistar雄性大鼠随机分为阴性对照组、实验组和干扰组。从建模第一周结束起,连续10周,每周一次,每组随机选取3只大鼠检测血液生化指标和尿液代谢指标。处死后,通过电子显微镜观察肾脏超微结构和病理组织学评估肾结石的形成情况。最后,采用蛋白质免疫印迹法检测肾组织中钙敏感受体(CaSR)和紧密连接蛋白14(Claudin-14)的表达。
与对照组相比,模型组大鼠肾脏大体结构发生改变。建模第四周时,纳米细菌组大鼠肾脏明显增大,肾体比增加,差异具有统计学意义(P<0.05)。肾脏剖面颜色变深,皮髓质结构欠清晰,在皮质髓质交界处可见淡黄色颗粒样物质堆积。纳米细菌组大鼠肌酐、尿酸、尿素氮及尿钙在第三周开始升高,第三周与第八周差异具有统计学意义(P<0.05)。但第八周后三组间差异无统计学意义。第四周时观察到结石形成,主要分布于肾小管及周围组织。纳米细菌组肾结石形成率为52.4%,干扰组为27.8%,差异具有统计学意义(P<0.05)。超微结构观察显示,从第四周起,纳米细菌组肾组织出现肾小管扩张、肾小管上皮细胞肿胀、颗粒样变性、脱落及淋巴细胞浸润,肾小管内可见少量钙盐结晶。第三周时,纳米细菌组CaSR和Claudin-14蛋白表达增加,差异具有统计学意义(P<0.05)。CaSR和Claudin-14表达与尿钙呈正相关(P<0.05)。
模型建立后第四周开始形成肾结石,结晶多位于肾小管。肾结石形成过程中,肾小管上皮细胞受损,表现为颗粒样变性及少量钙盐结晶,伴有部分肾小管开始扩张、上皮肿胀、颗粒样变性、坏死及脱落,肾间质淋巴细胞浸润,肾小管内可见少量钙盐结晶,且随时间加重。血清肌酐、血清尿酸、尿素氮及尿钙水平自第三周起随时间升高,第八周后恢复正常。CaSR和Claudin-14蛋白表达上调,且与24小时尿钙排泄值呈正相关。