联合膀胱灌注对大鼠间质性膀胱炎治疗研究

Study on the Treatment of Interstitial Cystitis in Rats with Combined In⁃travesical Instillation

目的通过膀胱灌注两种黏膜保护剂戊聚糖多硫酸钠(pentosanpolysulphate,PPS)及透明质酸联用替代葡萄糖氨基聚糖(glycosaminoglycan,GAG)层,研究大鼠膀胱间质炎模型中大鼠膀胱黏膜及黏膜下层肥大细胞(mastcells,MC)表达的变化情况。方法本研究选取2021年1-2月成年SD雌性大鼠30只,随机分为空白组,IC组,干预IC1组,干预IC2组,干预IC3组,每组6只。空白组:经尿道膀胱灌注生理盐水,不做任何处理。IC组:经尿道膀胱灌注硫酸鱼精蛋白、脂多糖、氯化钾溶液,排尽药物后用磷酸缓冲液(phosphatebufferedsa⁃line,PBS)冲洗。干预IC组:C1组(联合用药组)在IC组的基础上,拔除尿管4 h后应用透明质酸、PPS;C2组(透明质酸组)在IC组的基础上,拔除尿管4 h后应用透明质酸;C3组(戊聚多硫酸钠组)在IC组的基础上,拔除尿管4 h后应用PPS。1周后处死3组大鼠,取膀胱组织送病理。结果空白组膀胱黏膜和黏膜下层都无破损,IC组及IC干预组膀胱黏膜均可以看出明显损坏,且黏膜层存在连续性破损伴黏膜下层脱落的情况,组内大鼠可以明显看出膀胱壁组织存在大面积的出血、充血情况。空白组膀胱黏膜固有层(2.64±1.02)/HP逼尿肌(0.99±0.63)/HP、浆膜层(2.36±0.97)/HP与IC组相比MC数显著增加,差异有统计学意义(P<0.05)。空白组和干预C1组黏膜固有层[干预C1组(2.77±1.11)/HP]、逼尿肌[干预C1组(1.18±0.59)/HP]和浆膜层肥大细胞数[干预C1组(1.62±0.84)/HP]对比,差异无统计学意义(P>0.05)。对各组MC观察研究发现,空白组膀胱黏膜处有较少MC存在,IC组MC有明显增生出现,而不同方式干预的3个干预组膀胱黏膜也发生较明显的MC增生情况,且明显高于空白组,差异有统计学意义(P<0.05)。各组大鼠膀胱肌层处可见少量MC,各组对比差异无统计学意义(P>0.05)。结论GAG替代的黏膜保护剂能够降低大鼠的炎症反应,即减少大鼠膀胱黏膜及膀胱肌层MC。

Objective To replace the glycosaminoglycan(GAG)layer by intravesical infusion of two mucosal protec⁃tive agents,pentosan polysulphate(PPS)and hyaluronic acid,to study the model of interstitial inflammation in rats with large bladder changes in the expression of MC in the bladder mucosa and submucosa of rats.Methods In this study,30 adult female SD rats from January to February 2021 were randomly divided into blank group,IC group,in⁃tervention IC1 group,intervention IC2 group and intervention IC3 group,with 6 rats in each group.IC3 group(n=6).Blank group:transurethral bladder perfusion with normal saline without any treatment.IC group:protamine sulfate,lipopolysaccharide and potassium chloride solution were infused into the urinary bladder through urethra,and washed with phosphate buffer solution(PBS)after drug drainage.Intervention IC group:on the basis of IC group,group C1(combined medication group)was treated with hyaluronic acid and PPS 4 hours after catheter removal;group C2(hyaluronic acid group)was treated with hyaluronic acid 4 hours after catheter removal on the basis of IC group;on the basis of IC group,PPS was applied in C3 group(pentopoly sodium sulfate group)4 hours after catheter removal.One week later,the three groups of rats were sacrificed,and the bladder tissues were collected for pathol⁃ogy.Results There was no damage to the bladder mucosa and submucosa in the blank group,and significant damage to the bladder mucosa in the IC group and the IC intervention group,and there was continuous damage to the muco⁃sal layer and submucosa shedding,and large area of bleeding and hyperemia in the bladder wall tissue of the rats in the group could be obviously observed.The number of MC in bladder mucosa lamina proper(2.64±1.02)/HP,detru⁃sor muscle(0.99±0.63)/HP and serosal layer(2.36±0.97)/HP in blank group was significantly increased compared with IC group,and the difference was statistically significant(P<0.05).There was no statistically significant differ⁃ence in the number of MC in mucosa lamina propria[intervention group 1C(2.77±1.11)/HP],detrusor muscle[inter⁃vention group 1C(1.18±0.59)/HP]and serosal layer[intervention group 1C(1.62±0.84)/HP]between the blank group and intervention C1 group(P>0.05).The observation and study of MC in each group showed that there were less MC in the bladder mucosa of the blank group,and significant MC hyperplasia appeared in the IC group,while significant MC hyperplasia also occurred in the bladder mucosa of the three intervention groups with different intervention meth⁃ods,which was significantly higher than that of the blank group,the difference was statistically significant(P<0.05).A small amount of MC was found in the bladder muscle layer of rats in each group,and there was no statistically sig⁃nificant difference among all groups(P>0.05).Conclusion GAG-substituted mucosal protective agent can reduce the inflammatory response in rats,that is,reduce the MC in the bladder mucosa and bladder muscle layer of rats.