Human primary epithelial cells of renal pelvis was established to investigate the adherence of uropathogenic Escherichia coli (UPEC) to this cell line, in which the primary cell culture was performed by using cultiv...Human primary epithelial cells of renal pelvis was established to investigate the adherence of uropathogenic Escherichia coli (UPEC) to this cell line, in which the primary cell culture was performed by using cultivation of the normal epithelium of renal pelvis in keratinocyte serum free medium (K-SFM) with epidermal growth factor (EGF) and bovine pituitary extract (BPE). Both UPEC132 obtained from urine specimen of patients with pyelonephritis and the pilus-free representative strain E. coli K-12p678- 54 were used to study the adherence of these strains on human primary epithelial cells of renal pelvis. The UPEC adherence was performed with observation on the morphological changes of the adhered cells, while the adhesion rates and indices were calculated in different times of experiment. In addition, the virulence genes hly and cnfl of UPEC132 were detected by multiplex PCR assay. In this study, the human primary epithelial cells of renal pelvis was found to exhibit the character of the transitional epithelial cells. Compared with the control group, the adhesion rates and indices began to increase from 15 min of the experiment time and reached its peak in 120 min. The adhesion rate and index of UPEC132 to human primary epithelial cells of renal pelvis were 74.4% and 34.0 respectively. Many microscopic changes in the primary cells adhered with UPEC132 could be detected, such as rounding or irregularity in shape, unevenness in staining and the cytoplasmic and nuclear changes. It suggests that human primary epithelial cells of renal pelvis can be used for the experiment on UPEC adhesion, thus providing a basis for the further study on the pathogenesis of UPEC.展开更多
Adhesion of P-type and type-1 fimbriated uropathogenic E.coli(UPEC)to uroepithelial cells initiates urinary tract infections(UTIs).This research aimed to evaluate the capacities of selected cranberry polyphenols and t...Adhesion of P-type and type-1 fimbriated uropathogenic E.coli(UPEC)to uroepithelial cells initiates urinary tract infections(UTIs).This research aimed to evaluate the capacities of selected cranberry polyphenols and their microbial metabolites to inhibit such adhesion in vitro using a modified fluorometric method.Data showed that the inhibition capacity of myricetin increased with concentration and plateaued at 70%.It had IC50 values of 13.2μM against P-type E.coli and 5.50μM against type-1 E.coli.Quercetin showed similar anti-adhesion capacities to myricetin.Procyanidin A2 and B2 had weaker anti-adhesion activities than myricetin and quercetin,with maximal inhibition capacities of 20%-30%against UPEC.Hippuric acid,a major metabolite of cranberry polyphenols in human urine,showed a maximal inhibition of 20%at 558μM against type-1 E.coli adhesion,whereas no anti-adhesion activity against P-type E.coli was detected.The fractions of cranberry fruit powder enriched with proanthocyanidin polymers showed the highest anti-adhesion activities compared to the fractions enriched with anthocyanins,flavonols,or proanthocyanidin oligomers.Overall,the anti-adhesion activities of cranberry polyphenols and metabolites depend on their structures and the types of fimbriae on E.coli.展开更多
基金This work was supported by the grants from the National Natural Science Foundation of China (30470096).
文摘Human primary epithelial cells of renal pelvis was established to investigate the adherence of uropathogenic Escherichia coli (UPEC) to this cell line, in which the primary cell culture was performed by using cultivation of the normal epithelium of renal pelvis in keratinocyte serum free medium (K-SFM) with epidermal growth factor (EGF) and bovine pituitary extract (BPE). Both UPEC132 obtained from urine specimen of patients with pyelonephritis and the pilus-free representative strain E. coli K-12p678- 54 were used to study the adherence of these strains on human primary epithelial cells of renal pelvis. The UPEC adherence was performed with observation on the morphological changes of the adhered cells, while the adhesion rates and indices were calculated in different times of experiment. In addition, the virulence genes hly and cnfl of UPEC132 were detected by multiplex PCR assay. In this study, the human primary epithelial cells of renal pelvis was found to exhibit the character of the transitional epithelial cells. Compared with the control group, the adhesion rates and indices began to increase from 15 min of the experiment time and reached its peak in 120 min. The adhesion rate and index of UPEC132 to human primary epithelial cells of renal pelvis were 74.4% and 34.0 respectively. Many microscopic changes in the primary cells adhered with UPEC132 could be detected, such as rounding or irregularity in shape, unevenness in staining and the cytoplasmic and nuclear changes. It suggests that human primary epithelial cells of renal pelvis can be used for the experiment on UPEC adhesion, thus providing a basis for the further study on the pathogenesis of UPEC.
基金funded in part by University of Florida Research Foundation.TEM images were acquired by Nicole J.Machi and Rodolfo Alvarado in the Interdisciplinary Center for Biotechnology Research at University of Florida.
文摘Adhesion of P-type and type-1 fimbriated uropathogenic E.coli(UPEC)to uroepithelial cells initiates urinary tract infections(UTIs).This research aimed to evaluate the capacities of selected cranberry polyphenols and their microbial metabolites to inhibit such adhesion in vitro using a modified fluorometric method.Data showed that the inhibition capacity of myricetin increased with concentration and plateaued at 70%.It had IC50 values of 13.2μM against P-type E.coli and 5.50μM against type-1 E.coli.Quercetin showed similar anti-adhesion capacities to myricetin.Procyanidin A2 and B2 had weaker anti-adhesion activities than myricetin and quercetin,with maximal inhibition capacities of 20%-30%against UPEC.Hippuric acid,a major metabolite of cranberry polyphenols in human urine,showed a maximal inhibition of 20%at 558μM against type-1 E.coli adhesion,whereas no anti-adhesion activity against P-type E.coli was detected.The fractions of cranberry fruit powder enriched with proanthocyanidin polymers showed the highest anti-adhesion activities compared to the fractions enriched with anthocyanins,flavonols,or proanthocyanidin oligomers.Overall,the anti-adhesion activities of cranberry polyphenols and metabolites depend on their structures and the types of fimbriae on E.coli.