艾灸“相对穴”对大鼠脊髓损伤后神经源性膀胱及内质网应激途径的影响  

Effect of moxibustion at"oppositely-located points"on neurogenic bladder after spinal cord injury and endoplasmic reticulum stress pathway in rats

作　　者：魏炜 杨志新[1,2] 王天雨 崔陶陶 陈建双 张超[1,2,3] 李娜 任立群[1,2] WEI Wei;YANG Zhi-xin;WANG Tian-yu;CUI Tao-tao;CHEN Jian-shuang;ZHANG Chao;LI Na;REN Li-qun(Chengde Medical University,Chengde 067000,Hebei Province,China;Hebei Key Laboratory of Nerve Injury and Repair,Chengde 067000;Institue of Basic Medicine,Chengde Medical University)

机构地区：[1]承德医学院,河北承德067000 [2]河北省神经损伤与修复重点实验室,承德067000 [3]承德医学院基础医学研究所

出　　处：《中国针灸》2022年第4期413-418,共6页Chinese Acupuncture & Moxibustion

基　　金：河北省引进国外智力项目:2019YX027A;河北省科技厅高水平人才团队建设专项项目:199A7759H;河北省高等学校科学技术研究重点项目:ZD2019085;河北省引进留学人员资助项目:C20190174。

摘　　要：目的:观察艾灸相对穴“命门”“神阙”对骶上脊髓损伤(SCI)后神经源性膀胱大鼠后肢运动及膀胱功能的影响,探究其在内质网应激途径介导的膀胱组织凋亡中发挥的作用。方法:将28只雌性Wistar大鼠随机分为假手术组(8只)和造模组(20只)。造模组采用改良Allen′s法损伤大鼠T_(10)段脊髓制备SCI后神经源性膀胱模型,将造模成功的16只大鼠随机分为模型组(8只)和艾灸组(8只)。艾灸组大鼠于造模麻醉清醒后2 h予艾灸“神阙”“命门”干预,每穴每次2壮,隔日1次,共干预5次。比较各组大鼠干预后后肢运动功能(BBB)评分、尿流动力学指标(膀胱最大容量、漏尿点压力、膀胱顺应性),HE染色法观察膀胱组织形态学变化,Western blot法及实时荧光定量PCR法测定膀胱组织内质网应激相关基因[葡萄糖调节蛋白78(GRP78)、激活转录因子4(ATF4)、半胱氨酸天冬氨酸特异性蛋白酶12(Caspase-12)]蛋白及mRNA表达。结果:模型组大鼠膀胱移行上皮细胞排列紊乱,膀胱壁变薄,可见细胞空泡变性及中性粒细胞浸润;艾灸组大鼠膀胱移行上皮细胞较模型组层次清晰、连续,可见轻度细胞空泡变性及少量中性粒细胞浸润。与假手术组比较,模型组大鼠BBB评分降低(P<0.01),膀胱最大容量及膀胱顺应性增高(P<0.01),膀胱组织GRP78、ATF4、Caspase-12蛋白及mRNA表达增高(P<0.01)。与模型组比较,艾灸组大鼠BBB评分增高(P<0.01),膀胱最大容量及膀胱顺应性降低(P<0.01),膀胱组织GRP78、ATF4、Caspase-12蛋白及mRNA表达降低(P<0.01)。结论:艾灸“相对穴”可改善脊髓损伤后神经源性膀胱大鼠的排尿功能,减轻尿潴留,其机制可能与下调膀胱组织内质网应激通路中GRP78、ATF4、Caspase-12的表达,从而减轻膀胱组织细胞凋亡有关。Objective To observe the effect of moxibustion at oppositely-located points"Mingmen"(GV 4)and"Shenque"(CV 8)on the motor function of the hind limbs and bladder function in rats with neurogenic bladder after suprasacral spinal cord injury(SCI),so as to explore the effect of this therapy on bladder tissue apoptosis mediated by endoplasmic reticulum stress pathway.Methods Twenty-eight female Wistar rats were randomly divided into a sham-operation group(8 rats)and a model establishment group(20 rats).Using the modified Allen′s method,the spinal cord of T_(10) segment was injured to establish a neurogenic bladder model in the model establishment group.Sixteen rats were modeled successfully and then divided into a model group(8 rats)and a moxibustion group(8 rats).In the moxibustion group,2 h after consciousness regaining from modeling anesthesia,moxibustion was exerted at"Shenque"(CV 8)and"Mingmen"(GV 4),2 cones at each acupoint in one intervention.The intervention was administered once every two days and 5-time intervention was required totally.After intervention,Basso,Beattie and Bresnahan locomotor rating scale(BBB)score for the motor function of the hind limbs,and the urodynamics indexes(maximum bladder capacity,urine leakage pressure and bladder compliance)were compared among groups.HE staining method was adopted to observe the morphological changes of bladder tissue.With Western blot method and real-time PCR assay,the protein and mRNA expressions of the endoplasmic reticulum stress-related genes(glucose-regulated protein 78[GRP78],activating transcription factor 4[ATF4]and cysteinyl aspartate specific proteinase-12[Caspase-12])were determined.Results The transitional epithelial cells were arranged irregularly,the bladder wall was getting thinner,and the cellular vacuolar degeneration and neutrophil infiltration were found in the model group.Whereas,compared with the model group,in the moxibustion group,the arrangement of transitional epithelial cells was clear and continuous in layers,the cellular vacuolar deg

关 键 词：脊髓损伤 神经源性膀胱 艾灸 相对穴 内质网应激 细胞凋亡 

分 类 号：K245.8[历史地理—历史学]