姜黄素对慢性尿路感染大鼠TLR4/NF-κB通路及尿路上皮屏障影响

尤小明; 施安娜; 金百冶

doi:10.11847/zgggws1127780

姜黄素对慢性尿路感染大鼠TLR4/NF-κB通路及尿路上皮屏障影响

Effect of curcumin on TLR4/NF-κB pathway and urinary epithelial barrier in rats with chronic urinary tract infection

摘要

摘要:
目的探讨姜黄素对慢性尿路感染大鼠尿路上皮屏障及Toll样受体4（TLR4）/核因子 – κB（NF-κB）通路影响。
方法大鼠随机分为假手术组、模型组、阳性对照组、姜黄素（低、中、高）剂量组，每组8只；除假手术组外，其余各组0.1 mL大肠杆菌菌液（密度为1 × 10⁷个/mL）注射入尿道建立慢性尿路感染大鼠模型，阳性对照组大鼠灌胃3.67 g/kg三金片，姜黄素（低、中、高）剂量组分别灌胃（0.075、0.150、0.300）g/kg姜黄素，假手术组和模型组灌胃等体积生理盐水，连续14 d。灌胃后收集6 h尿液并量取体积；收集大鼠静脉血和膀胱组织，酶联免疫吸附（ELISA）检测血清中白介素（IL）-6、IL-8水平；计数尿白细胞；蛋白免疫印迹检测膀胱组织中TLR4、NF-κB、钙黏素（E-cadherin）、闭锁小带蛋白 – 1（ZO-1）水平。
结果与假手术组比较，模型组大鼠尿量（3.47 ± 0.65）vs（2.46 ± 0.35）mL、膀胱组织中E-cadherin（0.69 ± 0.07）vs（0.18 ± 0.02）、ZO-1（0.43 ± 0.04）vs（0.08 ± 0.02）蛋白水平降低（P < 0.05）；血清中IL-6（0.13 ± 0.03）vs（0.26 ± 0.05）、IL-8水平（0.53 ± 0.16）vs（1.32 ± 0.33）、尿白细胞数量（23.48 ± 3.58）vs（115.69 ± 14.89）、膀胱组织中TLR4（0.23 ± 0.04）vs（1.05 ± 0.11）、NF-κB（0.09 ± 0.01）vs（0.86 ± 0.09）蛋白表达升高（P < 0.05）。与模型组比较，姜黄素中剂量组大鼠膀胱组织中E-cadherin（0.35 ± 0.06）、ZO-1（0.35 ± 0.05）蛋白水平升高（P < 0.05）；中高剂量姜黄素组大鼠血清中IL-6（0.18 ± 0.02）、IL-8（0.68 ± 0.14）水平，尿白细胞数量（64.83 ± 7.43），膀胱组织中TLR4（0.35 ± 0.04）、NF-κB（0.33 ± 0.04）蛋白水平降低（P < 0.05）。
结论姜黄素对尿路感染大鼠具有一定保护作用，其机制可能与下调TLR4/NF-κB通路表达有关。

Abstract:
Objective To investigate the effect of curcumin on urinary barrier and Toll-like receptor 4 (TLR4)/nuclear factor-κB (NF-κB) pathway in rats with chronic urinary tract infection.
Methods Totally 48 male Sprague-Dawley rats were randomly divided into a sham operation group and a model group (with intragastric administration of saline), a positive drug group (with intragastric administration of Sanjin Tablet – a Chinese traditional medicine for treatment of urinary tract infections at the dosage of 3.67 g/kg), and low, moderate and high dose curcumin groups (with intragastric administration of curcumin at dosages of 0.075, 0.15, 0.3 g/kg). The treatments were conducted once a day continuously for 14 days. The chronic urinary tract infection model was established by urethra injection of 0.1 ml Escherichia coli suspension (concentration: 1 × 10⁷/ml) in all the rats except for those of sham operation group. By the end of last treatments, the rats′ urine in six hours, specimens of venous blood and bladder tissue were collected for detections of urine volume, urinary leukocyte count, serum interleukin (IL)-6 and IL-8 with enzyme-linked immunosorbent assay (ELISA), and tissular NF-κB/E-cadherin/zonula occludens protein 1 (ZO-1) with Western blot.
Results Compared with the sham-operation rats, the model rats had significantly decreased urine volume (2.46 ± 0.35 vs. 3.47 ± 0.65), serum IL-6 (0.13 ± 0.03 vs. 0.26 ± 0.05) and IL-8 (0.53 ± 0.16 vs. 1.32 ± 0.33), bladder tissular E-cadherin (0.18 ± 0.02 vs. 0.69 ± 0.07) and ZO-1 (0.08 ± 0.02 vs. 0.43 ± 0.04) (all P < 0.05) but significantly increased urinary leukocyte count (115.69 ± 14.89 vs. 23.48 ± 3.58) and bladder tissular TLR4 (1.05 ± 0.11 vs. 0.23 ± 0.04) and NF-κB (0.86 ± 0.09 vs. 0.09 ± 0.01) (P < 0.05 for all). Significantly increased bladder tissular E-cadherin (0.35 ± 0.06) and ZO-1 (0.35 ± 0.05) were detected in the rats with moderate curcumin but decreased urinary leukocyte count (64.83 ± 7.43), serum IL-6 (0.18 ± 0.02) and IL-8 (0.68 ± 0.14), and bladder tissular TLR4 (0.35 ± 0.04) and NF-κB (0.33 ± 0.04) in the rats with moderate and high curcumin in comparison with those in the model rats (P < 0.05 for all).
Conclusion Curcumin may has anti-inflammatory effect by down-regulating TLR4/NF-κB pathway in rats with bacteria- induced chronic urinary tract infection.

HTML全文

参考文献(20)

施引文献

资源附件(0)