aldh2基因表达对氯胺酮诱导的小鼠膀胱炎症及纤维化的影响

Effect of Aldh2 gene expression on Ketamine-induced bladder inflammation and fibrosis in mice

目的:探究乙醛脱氢酶2(aldh2)基因表达对氯胺酮诱导的小鼠膀胱炎症及纤维化的影响。方法:首先将8周龄ICR雄性野生型(WT)小鼠共20只随机分为氯胺酮(50 mg/kg)组和生理盐水组,分别诱导4周和8周。取出各组小鼠膀胱,运用实时荧光定量聚合酶链反应(qPCR)法检测各组WT小鼠aldh2 mRNA的表达情况。随后再选取ICR雄性aldh2基因敲除型(KO)与WT型小鼠各20只,分别随机分为生理盐水对照组和氯胺酮诱导组(50 mg/kg)。在第4周和8周时,取出膀胱组织用于检测其炎症和纤维化水平。结果:qPCR显示经氯胺酮处理后的WT小鼠膀胱aldh2 mRNA的相对表达量(1.84±0.24)显著高于生理盐水组(1.02±0.15)(P<0.01)。进一步在氯胺酮诱导组的比较中发现,KO小鼠较WT小鼠体重增加缓慢(P<0.01),并且KO炎症细胞因子白细胞介素1β(IL-1β)及6(IL-6)的表达显著增加(P<0.05);病理组织提示KO小鼠膀胱黏膜屏障消失、炎症细胞浸润、黏膜下层和肌层纤维化增加;蛋白质印迹(Western blot)结果也提示KO小鼠胶原蛋白COL-I的表达量(1.16±0.10)较WT小鼠(0.73±0.07)明显增加(P<0.01);而在生理盐水组中,KO与WT小鼠的炎症和纤维化水平比较,无统计学差异(P>0.05)。结论:aldh2基因可在氯胺酮相关性膀胱炎(KIC)的发生发展中发挥一定的抗炎抗纤维化的保护作用。

Objective: To investigate the effect of acetaldehyde dehydrogenase 2(aldh2)gene expression on bladder inflammation and fibrosis induced by ketamine in mice.Methods: Firstly,twenty 8-week-old ICR male wild type(WT)mice were randomly divided into ketamine(50 mg/kg)group and normal saline group,which were induced for 4 weeks and 8 weeks respectively.The bladder of each group of mice were taken out and the expression of aldh2 mRNA in WT mice of each group were detected by real-time fluorescence quantitative polymerase chain reaction(qPCR).Then 20 ICR male aldh2 knockout(KO)mice and 20 WTT mice were randomly divided into normal saline control group and ketamine induction group(50 mg/kg).At the 4 th and 8 th week,bladder tissues were removed to detect the level of inflammation and fibrosis.Results: qPCR showed that the relative expression of aldh2 mRNA(1.84±0.24)in the bladder of WT mice treated with ketamine was significantly higher than that in the saline group(1.02±0.15)(P<0.01).Futher comparison of ketamine-induced group showed that KO mice gained weight more slowly than WT mice(P<0.01),and the expression of inflammatory cytokines IL-1β and IL-6 in KO mice were significantly increased(P<0.05).Pathology showed that the bladder mucosal barrier of KO mice disappeared,inflammatory cells infiltrated,submucosa and muscle layer fibrosis increased.The results of Western blot also showed that the expression of collagen COL-I in KO mice(1.16 ± 0.10)was significantly higher than that in WT mice(0.73 ± 0.07)(P<0.01),but there was no significant difference in inflammation and fibrosis between KO mice and WT mice in normal saline group(P>0.05).Conclusion: Aldh2 gene may play a protective role in anti-inflammation and anti-fibrosis in the occurrence and development of in ketamine-associated cystitis(KIC).