基于SCF/C-kit通路探讨次髎穴注射甲钴胺对骶上脊髓损伤后神经源性膀胱大鼠的治疗机制  

Exploring the Therapeutic Mechanism of Injecting Mecobalamin into Ciliao(BL 32)on Neurogenic Bladder in Rats after Spinal Cord injury Based on the SCF/C-kit Pathway

作　　者：李卓伦 张润宁[2] 丁天红[2] 刘欣[1] 周孝敏 LI Zhuolun;ZHANG Running;DING Tianhong;LIU Xin;ZHOU Xiaomin(Shaanxi University of Chinese Medicine,Xianyang Shaanxi 712046,China;The Second Affiliated Hospital of Shaanxi University of Chinese Medicine,Xianyang Shaanxi 712046,China)

机构地区：[1]陕西中医药大学,陕西咸阳712046 [2]陕西中医药大学第二附属医院,陕西咸阳712046

出　　处：《中医药导报》2023年第12期24-28,共5页Guiding Journal of Traditional Chinese Medicine and Pharmacy

基　　金：陕西中医药大学校级科研课题立项项目(2021CX30);陕西省中医药管理局科研项目(JCPT040);咸阳市科学技术研究发展计划项目(2019k02-118);陕西省重点研发计划(2017SF-304)。

摘　　要：目的:探讨次髎穴注射甲钴胺对骶上脊髓损伤后神经源性膀胱大鼠的治疗作用及其对SCF/C-kit通路的影响。方法:36只SD大鼠按照随机数字表法分为假手术组、模型组、穴位注射组,每组12只。假手术组仅咬除椎板,不损伤脊髓;模型组和穴位注射组大鼠通过改良Allen’s法制作T10脊髓损伤大鼠模型。造模成功后,穴位注射组于次髎穴直刺15 mm注射甲钴胺,0.1 mL/次,1次/d,左右交替,共干预28 d。HE染色观察膀胱组织和肾脏组织形态学变化;采用RT-qPCR检测膀胱SCF mRNA、C-kit mRNA、JAK2 mRNA、STAT3 mRNA表达情况;采用Western blotting法检测膀胱组织SCF、C-kit、JAK2、STAT3蛋白表达情况。结果:与假手术组比较,模型组大鼠膀胱组织黏膜层可见上皮细胞变性脱落,胞质空泡化,固有层可见较多毛细血管淤血扩张,伴有少量淋巴细胞与中性粒细胞浸润,结缔组织排列疏松;肾皮质可见多处肾小管萎缩,伴有结缔组织增生及少量淋巴细胞浸润。模型组大鼠膀胱组织中SCF、C-kit、JAK2、STAT3的蛋白及mRNA相对表达量均明显高于假手术组(P<0.05)。与模型组比较,穴位注射组大鼠膀胱上皮细胞变性脱落减少,固有层局部可见水肿,少量淋巴细胞浸润;肾皮质中未见肾小管萎缩,未发现有结缔组织增生和淋巴细胞浸润。穴位注射组大鼠膀胱组织中SCF、C-kit、JAK2、STAT3的蛋白及mRNA相对表达量均明显低于模型组(P<0.05)。结论:次髎穴注射甲钴胺可改善骶上脊髓损伤后神经源性膀胱大鼠的膀胱组织损伤,其机制可能是降低了膀胱组织中SCF、C-kit、JAK2、STAT3的表达。Objective:To investigate the therapeutic effect of injecting mecobalamin into Ciliao(BL 32)on neurogenic bladder in rats with suprasacral spinal cord injury and its impact on the SCF/C-kit pathway.Methods:Totally 36 SD rats were divided into sham operation group,model group and acupoint injection group according to the method of random number table,with 12 rats in each group.The sham operation group only chewed off the vertebral lamina.The model of T10 spinal cord injury rats was made by modified Allen's method in the model group and acupoint injection group.After the successful modeling,the corresponding intervention was given.The acupoint injection group directly punctured Ciliao(BL 32)for 15 mm,with mecobalamin injection of 0.1 mL every time,once a day,alternately left and right,for 28 days.Histomorphological changes of bladder and kidney were observed by HE staining.The expression of SCF mRNA,C-kit mRNA,JAK2 mRNA,STAT3 mRNA in bladder was detected by RT-qPCR.The expression of SCF,C-kit,JAK2 and STAT3 protein in bladder tissue was detected by Western blotting.Results:According to the results of HE staining,compared with the sham operation group,the mucosa of bladder tissue in the model group showed degeneration and exfoliation of epithelial cells,vacuolization of cytoplasm,more congestion and expansion of capillaries in the lamina propria,with a small number of lymphocytes and neutrophils infiltrating,and loose connective tissue arrangement.The renal cortex of the model group showed multiple renal tubular atrophy,accompanied by connective tissue hyperplasia,and a small amount of lymphocyte infiltration.The model group showed higher relative expression of SCF,C-kit,JAK2,STAT3 protein and mRNA in the bladder tissue than sham operation group(P<0.05).Compared with the model group,the degeneration and abscission of bladder epithelial cells in the acupoint injection group were reduced,and edema was seen locally in the lamina propria.A small amount of lymphocytes were infiltrated in acupoint injection group.There was

关 键 词：脊髓损伤 穴位注射 次髎穴 甲钴胺 神经源性膀胱 大鼠 

分 类 号：R285.5[医药卫生—中药学]