电针气海、中极、关元穴改善压力性尿失禁尿道括约肌线粒体损伤机制研究

Effect of electroacupuncture at Qihai,Zhongji and Guanyuan points on mitochondrial damage of urethral sphincters in stress urinary incontinence

目的:观察电针对压力性尿失禁(SUI)环境下尿控的影响,并基于线粒体动力学探讨相关作用机制。方法:对SD大鼠进行随机分组,空白组未采取任何处理,假手术组完成手术解剖未干预,模型组行双侧卵巢切除+阴道扩张,假针组予非穴位电针,电针组进行穴位电针。尿动力学仪测量各组大鼠的最大膀胱容量(MBC)和腹腔漏尿点压力(ALPP)和漏尿点压力(LPP);HE染色观察尿道括约肌肌纤维形态改变;ELISA试剂盒检测线粒体膜电位、ATP和氧化应激相关指标;电镜观察尿道括约肌线粒体形态;Western blot检测尿道括约肌线粒体动力学相关因子线粒体融合素1(Mfn1)、Mfn2、线粒体动力相关蛋白1(Drp1)、线粒体分裂因子(MFF)、沉默信息调节因子1(SIRT1)和过氧化物酶体增殖物激活受体γ共激活因子-1α(PGC-1α)蛋白表达。结果:与假手术组比较,模型组大鼠尿控指标MBC、ALPP和LPP显著降低(均P<0.05),尿道括约肌纤维断裂、排列紊乱,线粒体明显肿胀,嵴增宽;线粒体膜电位、ATP、琥珀酸脱氢酶(SDH)和超氧化物歧化酶(SOD)水平显著降低(均P<0.05),丙二醛(MDA)水平显著升高(P<0.05);SIRT1、PGC-1α、Mfn1和Mfn2蛋白表达显著降低(均P<0.05),Drp1和MFF蛋白表达显著升高(均P<0.05)。与假针组比较,电针组大鼠的MBC、ALPP和LPP显著升高(均P<0.05),尿道括约肌纤维断裂、排列紊乱及线粒体的肿胀、嵴增宽情况显著改善;线粒体膜电位和ATP水平以及SDH和SOD活性显著升高(均P<0.05),MDA水平显著降低(P<0.05);SIRT1、PGC-1α、Mfn1、Mfn2蛋白表达显著升高(均P<0.05),Drp1和MFF蛋白表达显著降低(均P<0.05)。结论:电针可通过调控线粒体动力学改善SUI环境下尿道括约肌肌纤维形态和线粒体形态及功能障碍。

Objective:To observe the effect of electroacupuncture in urinary control and further explore the relevant mechanism based on mitochondrial dynamics.Methods:SD rats were randomly divided into groups.The control group did not take any treatment,the sham surgery group completed the surgical anatomy without intervention,the model group received bilateral ovariectomy+vaginal dilatation,the sham acupuncture group received non acupoint electroacupuncture,and electroacupuncture group receives acupoint electroacupuncture.The maximum bladder capacity(MBC),abdominal leak point pressure(ALPP) and leak point pressure(LPP) of rats in each group were measured by urodynamic testing.HE staining was used to observe the morphological changes of urethral sphincters muscle fibers.ELISA kit was used to detect mitochondrial membrane potential,ATP and oxidative stress related indicators.The morphology of mitochondria in urethral sphincters was observed by electron microscope.Western blot method was used to detect mitochondrial fusion protein 1(Mfn1),Mfn2,mitochondrial dynamics related protein 1(Drp1),mitochondrial fission factor(MFF),silent information regulator 1(SIRT1) and peroxisome proliferators-activated receptor γ coactivator-1α(PGC-1α) protein expression.Results:Compared with the sham surgery group,the urinary control indexes(MBC,ALPP and LPP) in the model group were significantly lower(all P<0.05),the urethral sphincters fibers were broken and disordered,the mitochondria were swollen and cristae widened,the mitochondrial membrane potential,ATP,succinate dehydrogenase(SDH) and superoxide dismutase(SOD) levels were significantly lower(all P<0.05),and the level of malondialdehyde(MDA) was significantly higher(P<0.05),the expression of SIRT1,PGC-1α,Mfn1 and Mfn2 proteins was significantly reduced(all P<0.05),while the expression of Drp1 and MFF proteins was significantly increased(all P<0.05).Compared with the sham acupuncture group,the MBC,ALPP and LPP of rats in the electroacupuncture group were significantly increased(all P<0.05),the fibers of the urethral sphincters were broken and disordered,and the swelling of mitochondria and the widening of crista were significantly improved;Mitochondrial membrane potential and ATP levels,SDH and SOD activities were significantly increased(all P<0.05),while MDA levels were significantly decreased(P<0.05),the expression of SIRT1,PGC-1α,Mfn1 and Mfn2 proteins significantly increased(all P<0.05),while the expression of Drp1 and MFF proteins significantly decreased(all P<0.05).Conclusion:Electroacupuncture can improve the fiber morphology and mitochondrial morphology and dysfunction of urethral sphincters under the SUI environment by regulating mitochondrial dynamics related factors.