益肾养髓方对脊髓型颈椎病大鼠脊髓中星形胶质细胞过度增殖的影响  

Effect of Yishen Yangsui Formula on Hyperproliferation of Astrocytes in Spinal Cord of Rats with Cervical Spondylotic Myelopathy

作　　者：秦晓宽 银河[1] 杨博文 金哲峰[1] 唐彬 刘志伟 陈忻[1] 朱立国[1,2] QIN Xiaokuan;YIN He;YANG Bowen;JIN Zhefeng;TANG Bin;LIU Zhiwei;CHEN Xin;ZHU Liguo(Wangjing Hospital of China Academy of Chinese Medical Sciences,Beijing 100102,China;Beijing Key Laboratory of Orthopedics of Traditional Chinese Medicine,Beijing 100007,China)

机构地区：[1]中国中医科学院望京医院,北京100102 [2]中医正骨技术北京市重点实验室,北京100007

出　　处：《中药新药与临床药理》2022年第2期173-179,共7页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然科学基金青年项目(8190152606);中国中医科学院中央级公益性科研院所基本科研业务费自主选题项目(ZZ11-082);中国中医科学院优秀青年科技人才(创新类)培养专项(ZZ14-YQ-022)。

摘　　要：目的观察益肾养髓方对脊髓慢性压迫大鼠脊髓(C;~C;)中胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)mRNA表达量及GFAP蛋白表达和分布的影响,以探讨益肾养髓方对脊髓型颈椎病(Cervical Spondylotic Myelopathy,CSM)大鼠脊髓中星形胶质细胞过度增殖和胶质瘢痕形成的作用。方法将56只健康雌性SD大鼠随机分为正常组、假手术组、模型组、阳性对照组及益肾养髓方低、中、高剂量组,每组8只。模型组、益肾养髓方各剂量组及阳性对照组大鼠采用吸水膨胀材料压迫脊髓建立脊髓型颈椎病大鼠模型,假手术组仅手术暴露脊髓。手术后2周,益肾养髓方不同剂量组采用中药水煎剂灌胃,阳性对照组采用单唾液酸四己糖神经节甘脂钠肌注,其余组采用生理盐水灌胃,每日1次,连续4周。于给药后第1、3、7、14、21、28天进行大鼠运动功能评价(BBB评分)。术后6周时HE染色观察脊髓组织结构,RT-qPCR法检测大鼠脊髓GFAP的mRNA表达,免疫荧光观察GFAP蛋白表达。结果灌胃后益肾养髓方各组及阳性对照组大鼠BBB评分明显高于模型组,差异具有统计学意义(P<0.05),说明益肾养髓方对改善脊髓型颈椎病大鼠脊髓功能有一定疗效。HE染色益肾养髓方各组及阳性对照组可见脊髓组织中瘢痕、空洞变少,胞浆逐渐增多,神经细胞形态接近假手术组。RT-qPCR及免疫荧光结果提示模型组GFAP mRNA及蛋白表达升高(P<0.05),益肾养髓方各组及阳性对照组可降低其表达,与模型组比差异具有统计学意义(P<0.05)。结论益肾养髓方可降低脊髓慢性压迫性损伤大鼠脊髓中GFAP的表达,从而减少胶质瘢痕的形成,促进大鼠后肢运动功能恢复。Objective To observe the effects of Yishen Yangsui formula on the expression of glial fibrillary acidic protein(GFAP)m RNA,and to study its effects on the expression and distribution of GFAP protein in the spinal cord(C;~C;) of rats with chronic spinal cord compression,further explore the effects of this formula on the hyperproliferation of astrocytes and the formation of glial scars in the spinal cord of cervical spondylotic myelopathy(CSM)rats.Methods Fifty-six healthy female SD rats were randomly divided into normal group,sham operation group,model group,low-,medium-and high-dose Yishen Yangsui formula group,and positive control group,with 8 rats in each group.All rats in the model group,traditional Chinese medicine group and positive control group were used to establish CSM model by compressing the spinal cord with water-absorbent swelling materials.The sham operation group only exposed the spinal cord.Two weeks after the operation,the different doses of Yishen Yangsui formula groups were given traditional Chinese medicine decoctions by gavage.Positive control group was given intramuscular injection of monosialotetrahexosylganglioside(dissolved in normal saline),and the rest groups were given intragastric administration of normal saline once a day for 4 consecutive weeks.The motor function of rats(BBB score)was evaluated at 1,3,7,14,21,28 days after the intervention.HE staining was performed to observe the tissue structure of the spinal cord at 6 weeks after operation,and RT-qPCR method was used to detect GFAP mRNA expression in rat spinal cord,immunofluorescence observation was used to investigate GFAP protein expression.Results After intragastric administration,the BBB scores of the rats in Yishen Yangsui formula groups and the positive control group were significantly higher than those in the model group,and the difference was statistically significant(P<0.05).The results indicated that the Yishen Yangsui formula has a certain effect on improving the spinal cord function of CSM rats.HE staining of Yishen Y

关 键 词：益肾养髓方 脊髓型颈椎病 慢性脊髓损伤 星形胶质细胞 过度增殖 胶质瘢痕 胶质纤维酸性蛋白(GFAP) 大鼠 

分 类 号：R285.5[医药卫生—中药学]