Escherichia coli Aggravates Calcium Oxalate Stone Formation via PPK1/Flagellin-Mediated Renal Oxidative Injury and Inflammation

Escherichia coli (E. coli) is closely associated with the formation of kidney stones. However, the role of E. coli in CaOx stone formation is not well understood. We explored whether E. coli facilitate CaOx stone formation and its mechanism. Stone and urine cultures were reviewed from kidney stone formers. The ability of calcium oxalate monohydrate (COM) aggregation was detected to evaluate the influence of uropathogenic E. coli, then gel electrophoresis and nanoLC-MS/MS to detect the crystal-adhered protein. Flagellin (Flic) and polyphosphate kinase 1 (PPK1) were screened out following detection of their role on crystal aggregation, oxidative injury, and inflammation of HK-2 cell in vitro. By transurethral injection of wild-type, Ppk1 mutant and Flic mutant strains of E. coli and intraperitoneally injected with glyoxylate in C57BL/6J female mice to establish an animal model. We found that E. coli was the most common bacterial species in patients with CaOx stone. It could enhance CaOx crystal aggregation both in vitro and in vivo. Flagellin was identified as the key molecules regulated by PPK1, and both of them could facilitate the crystal aggregation and mediated HK-2 cell oxidative injury and activated the inflammation-related NF-κB/P38 signaling pathway. Wild-type strain of E. coli injection significantly increased CaOx deposition and enhanced oxidative injury and inflammation-related protein expression, and this effect could be reversed by Ppk1 or Flic mutation. In conclusion, E. coli promotes CaOx stone formation via enhancing oxidative injury and inflammation regulated by the PPK1/flagellin, which activated NF-κB/P38 pathways, providing new potential drug targets for the renal CaOx calculus precaution and treatment.

Antiurolithiatic activity of Berberis asiatica by In vitro calcium oxalate crystallization methods

The primary objective of this research was to investigate the antiurolithiatic effect of the aqueous Heartwood extract of Berberis asiatica (AEBA) on in vitro crystallization methods. The antiurolithiatic behaviour was carried out in the presence and absence of AEBA at the concentration range of 100-1000 μg/ml by employing crystal nucleation, crystal aggregation, and crystal growth assay methods. Standard drug Cystone was made use of positive control in the concentration range of 100-1000 μg/ml. Inhibition efficiency of AEBA on crystal nucleation, crystal aggregation and crystal growth was spectrophotometrically validated. The percentage inhibition rate of crystal nucleation, crystal aggregation and crystal growth by AEBA and standard drug cystone was endorsed to be dose-dependent in nature. The half maximal inhibitory concentration (IC50) values of standard drug cystone on crystal nucleation, crystal aggregation and crystal growth were estimated to be 415.30±21.35, 573.7±65.53 and 566.20±62.06 μg/ml, respectively, while the AEBA, IC50 values were reckoned to be 839±69.13, 927.10±69.98 and 851±86.60 μg/ml, respectively. The findings of in vitro crystallization study disclosed that an aqueous Heartwood extract of Berberis asiatica possesses calcium oxalate crystal inhibition activity on crystal nucleation, crystal aggregation, and crystal growth recommended it as a potent and promising antiurolithiatic activity.

Crystal aggregation into periodically grating-banded assemblies in phthalic acid modulated by molten poly(ethylene oxide)

A small-molecule compound, phthalic acid (PA), crystallized in the presence of poly(ethylene oxide) (PEO) with various compositions was utilized as a model to investigate the morphology and crystal assembly of periodically ordered structures in banded spherulites.

ROLE OF TAMM-HORSFALL PROTEIN (THP) IN LITHOGENIC PROCESS : ANIMAL STUDIES

One of the defenses against nephrolithiasis is provided by macromolecules that modulate the nucleation, growth, aggregation and retention of crystals in the kidneys.According to its well-known physico-chemical properties.THP has a dual role in modifying crystal aggregation: at high pH and low ionic strength (IS),THP is a powerful crystal aggregation inhibitor.Upon lowering pH and rasing ionic strength THP viscosity increases,leading to reduced crystal aggregation inhibition.For this purpose eight guinea pigs were made hyperoxaluric.The treatment was given for fifteen days;then urine samples were collected before treatment ;then on 5,10,15and 25 day( after treatment) and in vitro addition of THP on 30th day which was isolated from hyperoxaluric and normal animals.The effect of EG+GM on urinary oxalate,THP and TBAR levels increases but after treatment the urine chemistry revert to normal profile,though plasma TBAR levels were appreciably high.The crystallization of calcium was almost double when THP was isolated from hyperoxaluric animals rather than normals.Our study suggested that THP act as a promoter