芍药甘草汤灌肠治疗大鼠氯胺酮相关性膀胱炎的实验研究

Effect of enema of Shaoyao Gancao Decoction in ketamine cystitis rats

关键词:  芍药甘草汤  氯胺酮相关性膀胱炎  氧化应激  炎症因子

Key words:  Shaoyao Gancao Decoction  ketamine cystitis  oxidative stress  inflammatory factor

摘要:

[目的] 探讨芍药甘草汤灌肠治疗氯胺酮相关性膀胱炎（KC）模型大鼠的疗效及其作用机制。[方法] 选取SD雄性大鼠40只，随机分为空白组、模型组、左氧氟沙星组、芍药甘草汤组，每组10只。采用氯胺酮连续腹腔注射12周建立KC模型。芍药甘草汤组给予芍药甘草汤水煎剂灌肠治疗，以左氧氟沙星为阳性对照组，给予左氧氟沙星灌肠治疗，空白组给予等量生理盐水灌肠处理，每日1次，持续4周。治疗结束行膀胱尿动力学检测，比较各组大鼠膀胱残余尿量、最大膀胱容量、最大膀胱压力及膀胱顺应性变化情况；疼痛检测比较各组大鼠疼痛阈值；采用苏木精-伊红染色法和酶联免疫吸附法（ELISA）检测各组大鼠膀胱组织病理学改变及血清中白介素-1β（IL-1β）、白介素-6（IL-6）及肿瘤坏死因子-α（TNF-α）的表达变化，蛋白质免疫印迹法检测大鼠膀胱组织中核因子-κB p65（NF-κB p65）、核因子E2相关因子2（Nrf2）、环氧化酶-2（COX-2）蛋白表达水平，TransAMTM ELISA法检测NF-κB p65、Nrf2的DNA结合活性。[结果] 与模型组相比，芍药甘草汤组大鼠膀胱最大容量、残余尿量及膀胱顺应性明显降低，最大膀胱内压明显升高，疼痛阈值明显高于模型组，差异有统计学意义（P<0.05）。芍药甘草汤组大鼠膀胱组织病理损伤明显改善，大鼠血清中IL-1β、TNF-α、IL-6炎症因子的表达明显降低，大鼠膀胱组织中NF-κB p65、COX-2的表达明显降低，Nrf2的蛋白表达显著上调，同时降低了NF-κB p65的活性，增高了Nrf2活性，差异有统计学意义（P<0.05）。[结论] 芍药甘草汤能够通过调控NF-κB/COX-2/Nrf2信号通路抑制KC模型大鼠氧化应激及炎症反应，改善膀胱功能，减轻膀胱组织损伤，从而起到治疗KC的作用。

Abstract:

[Objective] To investigate the effect and mechanism of Shaoyao Gancao Decoction enema on ketamine associated cystitis(KC) model rats. [Methods] Forty male SD rats were randomly divided into blank group,model group,levofloxacin group and Shaoyao Gancao Decoction group,with 10 rats in each group. KC model was established by intraperitoneal injection of ketamine for 12 weeks. After successful modeling,Shaoyao Gancao Decoction group was treated by Enema with 5 mL/kg,the positive control group was given levofloxacin,once a day for four weeks. Bladder urodynamic test was performed to compare the changes of residual urine volume,maximum bladder volume,maximum bladder pressure and bladder compliance in each group;pain threshold was detected and compared;He staining and ELISA were used to detect the pathological changes of bladder tissue and the levels of IL-1 β,IL-6 and TNF-α in serum. Western-blot was used to detect the protein expression of NF-κB p65,Nrf2 and COX-2 in bladder tissue. TransAMTM ELISA was used for the detection of DNA binding activity of NF-κB p65 and Nrf2. [Results] Compared with the model group,the maximum bladder volume,residual urine volume and bladder compliance in the Shaoyao Gancao Decoction group were significantly decreased,the maximum bladder internal pressure was increased,and the pain threshold was higher than model group (P<0.05). And the pathological damage of bladder tissue was improved,the expression of IL-1 β,TNF-α,IL-6 in serum and NF-κB p65,COX-2 protein was decreased,the expression of Nrf2 was up-regulated,while NF-κB p65 activity was decreased and Nrf2 activity was increased (P<0.05). [Conclusion] Shaoyao Gancao Decoction can inhibit oxidative stress and inflammatory reaction in KC model rats by regulating NF-KB/COX-2/Nrf2 signaling pathway,improve bladder function and reduce bladder tissue damage.