环孢素A在透明质酸诱导大鼠间质性膀胱炎/膀胱疼痛综合征中的治疗作用

Therapeutic effect of cyclosporin A on hyaluronidase- induced interstitial cystitis/bladder pain syndrome(IC/BPS)in rats

目的观察环孢素A（Cs A）在透明质酸酶诱导的大鼠间质性膀胱炎/膀胱疼痛综合征（IC/BPS）中的治疗作用。方法将45只成年雌性Sprague-Dawley（SD）大鼠（200-250 g）,分为对照组15只（膀胱灌注生理盐水）,模型组15只（膀胱灌注透明质酸酶）,Cs A治疗组15只（膀胱灌注透明质酸酶＋Cs A）。采用长期（1个月）间歇灌注透明质酸酶（4 g/L）构建大鼠IC/BPS模型,尿流动力学检测膀胱功能,Von Frey刷检测泌尿生殖区疼痛变化,硝酸还原酶法测定膀胱组织NO的含量,逆转录聚合酶链反应（RT-PCR）检测i NOS、IL-6、IL-10及IL-17 m RNA表达,ELISA法检测膀胱组织IL-6、IL-10及IL-17含量。结果与对照组相比,模型组膀胱大鼠膀胱i NOS、IL-6、IL-10、IL-17 m RNA表达均显著升高（P〈0.001）,在予以Cs A治疗后,其m RNA表达均显著下调（P〈0.05）。模型组膀胱NO含量为9.73±2.62μmol/g;IL-6含量为125.4±11.25μg/g;IL-10含量为64.05±6.26μg/g;IL-17含量为90.61±10.49μg/g;排尿时间间隔为148.23±40.75 s;膀胱容量为0.41±0.06 m L。对照组膀胱NO含量为1.31±0.55μmol/g;IL-6含量为55.18±5.07μg/g;IL-10含量为32.12±3.82μg/g;IL-17含量为44.45±4.92μg/g;排尿时间间隔为441.90±34.96 s;膀胱容量为1.27±0.10 m L。与对照组比较,模型组大鼠膀胱NO、IL-6、IL-10、IL-17含量均明显升高,排尿时间间隔缩短,膀胱容量降低,疼痛评分显著增加（P〈0.05）。与模型组相比,Cs A治疗组大鼠的膀胱组织NO、IL-6、IL-10、IL-17含量均显著减少（P〈0.001）,分别为3.97±1.71μmol/g;62.29±6.68μg/g;33.51±5.77μg/g;51.88±6.67μg/g,排尿时间间隔及膀胱容量明显增加（P〈0.05）,分别为422.06±42.22 s、1.14±0.15 m L,疼痛评分也明显下调（P〈0.05）。结论在透明质酸酶诱导的IC/BPS大鼠模型中,Cs A膀胱灌注治疗能显著改善大鼠的尿流动力学及疼痛症状,这可能与其下调i NOS、NO、IL-6、IL-10及IL-17等炎症介质的表达有关。

Objective To evaluate the therapeutic effect of cyclosporin A（Cs A）on hyaluronidaseinduced interstitial cystitis/bladder pain syndrome（IC/BPS）in rats. Methods Forty- five Sprague-Dawley rats were randomized into control group（15 rats,intravesical normal saline）,model group（15rats,intravesical hyaluronidase）and Cs A- treated group（15 rats,intravesical hyaluronidase ＋ Cs A）.Voiding patterns were investigated by cystometrography. The changes of pain in urogenital area were detected by Von-Frey Hairs. The NO level of bladder tissue were measured by using nitric acid deoxidizeenzyme method. The m RNA 1evels of i NOS,IL-6,IL-10,IL-17 were analyzed with real time reverse transcription polymerase chain reaction. The IL- 6,IL- 10 and IL- 17 level were measured by ELISA.Results Compared with control,the m RNA level of i NOS,IL-6,IL-10,IL-17 increased significantly（all P values 〈0.001）. The Cs A-treated group showed decreased m RNA levels of i NOS,IL-6,IL-10,IL-17（all P values〈 0.05）. In model group,the NO level was（9.73±2.62）μmol/g;the IL-6 level was（125.4±11.25）μg/g;the IL-10 level was（64.05±6.26）μg/g;the IL-17 level was（90.61±10.49）μg/g. The intercontraction intervals was（148.23±40.75）s;The bladder capacity was 0.41±0.06 ml. In control group,the NO level was 1.31±0.55 μmol/g;the IL-6 level was 55.18±5.07 μg/g. The IL-10 level was（32.12±3.82）μg/g;the IL-17 level was（44.45±4.92）μg/g. The intercontraction intervals was 441.90±34.96 s;The bladder capacity was 1.27±0.10 m1. Compared with control,the rats of model group showed high NO,IL-6,IL-10 and IL-17 level,decreased intercontraction intervals and bladder capacity,high pain score（all P〈0.05）. After administration of Cs A,the NO,IL-6,IL-10 and IL-17 level of bladder reduced（P〈0.001）,which was 3.97 ± 1.71 μmol/g,62.29 ± 6.68 μg/g,33.51 ± 5.77 μg/g and 51.88 ± 6.67 μg/g,respectively. Both intercontraction intervals and bladder capacity increased significantly（both P〈0.05）.And they were 422.06±42.22 s and 1.14±0.15 ml,respectively. The pain score also decreased significantly（P〈0.05）. Conclusion In the IC/BPS rat model induced by hyaluronidase,intravesical Cs A can significantly ameliorate the frequent urination and bladder pain,which may be related to the down-regulation of i NOS,NO,IL-6,IL-10 and IL-17.