间歇性低氧对膀胱逼尿肌细胞凋亡的影响及益智仁调节机制  

Effect of intermittent hypoxia on bladder detrusor cell apoptosis and regulatory mechanism of Alpiniae oxyphyllae Fructus

作　　者：苏苗赏[1] 薛思忱 徐莉[1] 任锡凯 黄楠[1] 唐祝亲 徐漫欢 SU Miao-shang;XUE Si-chen;XU Li;REN Xi-kai;HUANG Nan;TANG Zhuqin;XU Man-huan(The Second Affiliated Hospital and Yuying Children's Hospital,The Second Medical College,Wenzhou Medical University,Wenzhou 325027,China;College of Laboratory Medicine and Life Science,Wenzhou Medical University,Wenzhou 325035,China)

机构地区：[1]温州医科大学附属第二医院育英儿童医院、第二临床医学院,浙江温州325027 [2]温州医科大学检验医学院、生命科学学院,浙江温州325035

出　　处：《中国病理生理杂志》2020年第12期2212-2219,共8页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81870075);浙江省自然科学基金资助项目(No.LY17H010003)。

摘　　要：目的:观察间歇性低氧(IH)对人膀胱逼尿肌细胞凋亡和钙离子通道的影响,并初步探讨中药益智仁(AOF)的调节机制。方法:建立人膀胱逼尿肌细胞IH模型,IH模式为5%O260 min-20%O230 min,6个循环。体外培养人膀胱逼尿肌细胞,随机分为6组,每组设8孔,分别为P2X3受体拮抗剂+IH组(A组)、M3受体拮抗剂+IH组(B组)、β3受体拮抗剂+IH组(C组)、AOF+IH组(D组)、生理盐水+IH对照组(NC组)和空气模拟对照(AC)组。干预结束后普通光镜下细胞板计数比较各组细胞密度,免疫荧光法观察细胞形态改变,流式细胞术检测细胞凋亡,膜片钳法测定细胞钙离子表达变化。结果:(1)与AC组细胞比较,NC组细胞密度增高、反应活跃,部分细胞出现突起、变圆,细胞边界模糊;A组和D组细胞密度较NC组有显著减低(P<0.05);(2)免疫荧光检测细胞α-肌动蛋白(α-SMA)表达情况:与AC组比较;NC组的平均吸光度(MA)显著增高(F=3.25,P<0.05);A组、D组细胞分别与NC组比较均有不同程度显著降低(P<0.05)。(3)与AC组细胞比较,NC组细胞凋亡率显著减低(P<0.05);A组与D组的细胞凋亡率分别与NC比较有显著增高(P<0.05);(4)与AC组细胞比较,NC组细胞钙离子通道表达显著减低(P<0.05);与NC组细胞比较,AOF组(100 mg/L)和AOF组(50 mg/L)钙离子通道表达显著增高(P均<0.05)。结论:IH可通过P2X3膀胱神经受体调节逼尿肌细胞增殖和凋亡,中、高剂量AOF可改变钙离子通道并对IH诱导的细胞损害起保护作用。AIM:To investigate the effect of intermittent hypoxia(IH)on bladder detrusor cells apoptosis and calcium channel,and to discuss the regulatory mechanism of Alpiniae oxyphyllae Fructus(AOF).METHODS:IH model of bladder detrusor cells was established by treating the cells with 6 cycles of 5%O2 for 60 min and 20%O2 for 30 min.Human bladder detrusor cells were cultured in vitro,randomly divided into 6 groups,each group had 8 holes.P2X3 receptor antagonist+IH(A)group,M3 receptor antagonist+IH(B)group,β3 receptor antagonist+IH(C)group,AOF+IH(D)group,saline+IH control(NC)group and air simulation control(AC)group were set up.The cells density and morphology were identified by the methods of counting chamber and immunofluorescence light microscopy(LM)after interventions.The apoptosis was analyzed by flow cytometry.Calcium channel expression was detected by patch clamp.RESULTS:(1)Compared with the cells in AC group,the cells density and activity were significantly increased in NC group(P<0.05);some cells appeared protrusions,turned round and blur in cell borders.(2)The results of immunofluorescence for detectingα-SMA protein expression showed that,compared with the cells in group AC,the mean absorbance(MA)in group NC was significantly increased(F=3.25,P<0.05);compared with the cells in group NC,that in group A and group D was both decreased significantly(P<0.05).(3)Compared with the cells in group AC,the apoptotic rate was significantly decreased in group NC(P<0.05);Compared with the cells in group NC,the apoptotic rates in group A and group D were both significantly increased(P>0.05).(4)Compared with the cells in group AC,calcium ion channel expression was significantly decreased(P<0.05).Compared with the cells in group NC,calcium ion channel expression in AOF(100 mg/L)and AOF(50 mg/L)group was significantly increased(P<0.05).CONCLUSION:IH regulates bladder detrusor cells proliferation and apoptosis through P2X3 bladder nerve receptors,high or moderate dose of AOF may change calcium channel and play a protective role in

关 键 词：间歇性低氧 膀胱逼尿肌 细胞凋亡 钙离子通道 

分 类 号：R563[医药卫生—呼吸系统] R363.2[医药卫生—内科学]