格列本脲对大鼠膀胱炎性衰老的影响及其可能的机制研究

Effect of Glyburide on Inflamm-aging Bladder of Rats and Underlying Mechanism

目的:探讨格列本脲对衰老膀胱炎症的影响及其可能机制。方法:将SD大鼠根据月龄随机分为3组:2月龄青年组、24月龄衰老对照组和24月龄衰老格列本脲组(10 mg·kg^(-1)·d^(-1),口服1个月),每组10只。采用HE染色观察膀胱形改变,天狼猩红染色检测胶原沉积,尿动力学测定膀胱功能,Western Blot检测膀胱组织中NOD样受体家族含pyrin结构域蛋白3(NLRP3)和白细胞介素-1β(IL1β)分子表达和衰老相关蛋白P16表达,免疫荧光检测α-SMA在膀胱黏膜下血管中的表达,免疫组化检测NLRP3和IL1β表达。结果:衰老对照组和衰老格列本脲组的NLRP3和IL1β表达量较青年组增加,而衰老格列苯脲组增加的程度低于衰老对照组(P <0. 05)。HE和天狼腥红染色结果显示衰老对照组和衰老格列本脲组膀胱黏膜和平滑肌层结构重塑,胶原沉积明显增加,且衰老格列本脲组的胶原蛋白肌纤维面积比衰老对照组减少(P <0. 05)。衰老对照组和衰老格列本脲组中膀胱黏膜下血管壁结构形态紊乱,管腔较青年组狭窄。衰老相关蛋白p16在衰老对照组和衰老组中较青年组显著增加(P <0. 05)。尿动力学未检测到明显的不稳定收缩,但衰老对照组排尿时间延长,残余尿量较青年组和衰老格列苯尿组增加。结论:格列本脲可以一定程度缓解衰老膀胱功能,作用机制可能与抑制衰老膀胱中NLRP3/IL-1β炎症小体活化相关。

Objective:To investigate the inhibitory effect of glyburide on the activations of NLRP3 inflammasome in aging bladder to relieve lower urinary tract symptoms.Methods:SD rats were divided into 3 groups according to the age:2-month-old young group,24-month-old senile control group and 24-month-old senile+glibenclamide group(10 mg·kg^-1·d^-1,oral administration for one month)with 10 rats in each.HE staining was used to observe the change of bladder remolding.Sirius red staining was used to detect collagen deposition.Urodynamics was used to detect bladder function.Western blot was used to detect the expression of NLRP3/IL-1βinflammatsome and senescence-associated protein P16 in bladder tissue.Immunofluorescence was used to detect the expression ofα-SMA in bladder submucous vessels.Immunohistochemistry was used to detect the expressions of NLRP3 and IL-1β.Results:The expressions of NLRP3 and IL-1βin the senile control group and the senile+glibenclamide group were higher than those in the young group,and the extent of expression increase in the senile+glibenclamide group was lower than that in the senile control group(P<0.05).HE and sirius red staining showed that the bladder mucosa and the collagen deposition were significantly increased in the senile control group and the senile+glibenclamide group,and the area of collagen fibers in the senile+glibenclamide group was less than that in the senile control group(P<0.05).In the aging control group and the senile+glibenclamide group,the structure of submucosal blood vessel wall was disordered,and the lumen was narrower than that in the young group.The aging-related protein p16 was significantly increased in the aging control group and the senile+glibenclamide group when compared with that in the young group(P<0.05).Urodynamics showed no significantly unstable contraction,while the urination time was prolonged in the senile control group,and the residual urine volume in the senile control group was higher than that in the young group and the senile+glibenclamide group.Conclusion:Glyburide can inhibit the activation of NLRP3 inflammasome in aging bladder and relieve bladder overactivity to a certain extent,and its mechanism may be related to the activation inhibition of NLRP3/IL-1βinflammasome in aging bladder.