脊髓康对大鼠脊髓损伤区小胶质细胞/神经元的影响

doi:10.12092/j.issn.1009-2501.2020.02.006

中国临床药理学与治疗学 ›› 2020, Vol. 25 ›› Issue (2): 151-158.doi: 10.12092/j.issn.1009-2501.2020.02.006

脊髓康对大鼠脊髓损伤区小胶质细胞/神经元的影响

潘娅岚^1,3，苑文超³，马勇^2,3，郭杨^2,3，郑苏阳³，徐桂华¹，刘家欢³

¹南京中医药大学中西医结合护理研究所，南京 210023，江苏； ²南京中医药大学附属医院骨伤科，南京 210029，江苏； ³南京中医药大学骨伤研究所，南京 210023，江苏

收稿日期:2019-10-08 修回日期:2019-12-19 出版日期:2020-02-26 发布日期:2020-03-06
通讯作者: 郭杨，男，博士研究生，副教授，研究方向：中医药治疗骨关节病。 Tel: 13451918933 E-mail：drguoyang@126.com
作者简介:潘娅岚，女，博士研究生，助理研究员，研究方向：中医药治疗骨关节病。 Tel: 025-85811654 E-mail：yalan_pan@sina.com
基金资助:
国家自然科学基金青年基金项目（81704100）；国家自然科学基金面上项目（81573997）；江苏高校“青蓝工程”资助项目（苏教师〔2018〕12号）； 2017年地方高校国家级大学生创新训练计划（201710315004）；江苏省高校优势学科南京中医药大学护理学资助项目（苏政办发2014（37）号）

Effect of JiSuiKang on the microglia infiltration and neuron survival in the spinal cord injury area of rats

PAN Yalan ^1,3, YUAN Wenchao³, MA Yong ^2,3, GUO Yang ^2,3, ZHENG Suyang ³, XU Guihua ¹, LIU Jiahuan ³

¹ Institute of Integrated Traditional Chinese and Western Medicine of Nanjing University of Chinese Medicine, Nanjing 210023, Jiangsu, China; ² Department of Orthopedics of Jiangsu Province Traditional Chinese Medicine Hospital, Affiliated to Nanjing University of Chinese Medicine, Nanjing 210029, Jiangsu, China; ³Institute of Traumatology and Orthopedics, Nanjing University of Chinese Medicine, Nanjing 210023, Jiangsu, China

Received:2019-10-08 Revised:2019-12-19 Online:2020-02-26 Published:2020-03-06

摘要/Abstract

摘要： 目的:观察中药复方脊髓康（JSK）对大鼠脊髓损伤区域小胶质细胞浸润与神经元存活情况的影响。方法:60只SPF级SD大鼠，随机分为假手术组（Sham）、模型组（Ctrl）、JSK低、中、高剂量组（JSK-L、JSK-M、JSK-H）、强的松组；改良Allen's法建立大鼠脊髓损伤（spinal cord injury,SCI）模型；术后各组以不同的方式干预，行大鼠双后肢运动功能评分（BBB评分）；酶联免疫吸附法（ELISA）检测血清炎症因子IL-1表达，损伤区脊髓组织行冰冻切片，CD11b/c、β3-Tubulin免疫荧光染色，观察小胶质细胞的浸润情况与神经元的生长情况。结果:在BBB评分方面：Sham、JSK-L、JSK-M、JSK-H、强的松组均高于Ctrl（P＜0.05）；在ELISA检测方面，Sham、JSK-L、JSK-M、JSK-H、强的松组的IL-1表达均低于Ctrl（P＜0.05），强的松组、JSK-H低于JSK-L、JSK-M（P＜0.05）；在小胶质细胞浸润百分比方面，JSK-L、JSK-M、JSK-H、强的松组均低于Ctrl（P＜0.05），JSK-H与强的松组相比差异无统计学意义（P＞0.05）。从神经元的生长状态上：JSK-L、JSK-M、JSK-H、强的松组优于Ctrl（P＜0.01）；JSK-H优于JSK-L、JSK-M（P＜0.05）。结论:中药复方JSK 抑制SCI后体内小胶质细胞的迁移浸润，促进受损神经元的恢复，进而促进SCI大鼠双下肢运功功能的恢复。

关键词: 脊髓康, 脊髓损伤, 小胶质细胞, 神经元, 迁移, 浸润

Abstract: AIM: To observe the effect of Chinese medicine compound JiSuiKang on the microglia infiltration and neuron survival at the spinal cord injury area of rats. METHODS: Sixty SD rats were randomly divided into sham operation group (Sham), model group (Ctrl), JSK low (JSK-L), JSK medium (JSK-M) and JSK high (JSK-H) dose group, prednisone group. The model of rat spinal cord injury (SCI) was established by improved Allen's method. After the operation, each group was intervened in different ways, and then we assessed the double hind limb motor function score in rats (BBB score). The heart perfusion was done and the spinal cord tissues from the injury area were taken. ELISA tests were used to detect the expression of inflammatory factors. The spinal cord tissue was sliced into sections and then immunofluorescence stained of CD11b/c and beta 3-Tubulin to observe the infiltration of microglia and the growth of neurons under the inverted fluorescence microscope. RESULTS:The BBB score of Sham, JSK-L, JSK-M, JSK-H and prednisone group were higher than Ctrl（P＜0.05）. For the expression of inflammation, the expressions of IL-1 in group Sham, JSK-L, JSK-M, JSK-H and prednisone were lower than that of Ctrl (P＜0.05). For the percentage of microglia infiltration, JSK-L, JSK-M, JSK-H and prednisone were all lower than that of Ctrl (P＜0.05), and there was no significant difference between JSK-H and prednisone(P＞0.05). For the growth state of the neuron, the JSK-L, JSK-M, JSK-H, prednisone group were better than Ctrl (P＜0.01), and JSK-H was better than JSK-L and JSK-M (P＜0.05). CONCLUSION: The Chinese medicine compound JSK inhibits the migration and invasion of microglia in SCI, promotes the recovery of damaged neurons and motor function in SCI rats.

Key words: JiSuiKang, spinal cord injury, microglia, neurons, migration, infiltration

中图分类号:

潘娅岚，苑文超，马勇，郭杨，郑苏阳，徐桂华，刘家欢. 脊髓康对大鼠脊髓损伤区小胶质细胞/神经元的影响[J]. 中国临床药理学与治疗学, 2020, 25(2): 151-158.

PAN Yalan, YUAN Wenchao, MA Yong, GUO Yang, ZHENG Suyang, XU Guihua, LIU Jiahuan. Effect of JiSuiKang on the microglia infiltration and neuron survival in the spinal cord injury area of rats[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2020, 25(2): 151-158.

参考文献

［1］Furlan JC, Sakakibara BM, Miller WC, et al. Global incidence and prevalence of traumatic spinal cord injury［J］. Can J Neurol Sci, 2013, 40 (4): 456-464.
［2］Wang HF, Yin ZS, Chen Y. Epidemiological features of traumatic spinal cord injury in Anhui province ［J］. Spinal Cord, 2013, 51(1): 20-22.
［3］Berkovitch Y, Cohen T, Peled E, et al. Hydrogel composition and laser micro‐patterning to regulate sciatic nerve regeneration［J］. J Tissue Engin Reg Med, 2017, 24(2): 58-62.
［4］Colonna M, Butovsky O. Microglia function in the central nervous system during health and neurodegeneration［J］. Annu Rev Immunol, 2017, 35: 441-468.
［5］Liu G, Fan GT, Guo GD, et al. FK506 attenuates the inflammation in rat spinal cord injury by inhibiting the activation of NF-κB in microglia cells［J］. Cell Mol Neurobiol, 2017, 37(5): 843-855.
［6］Fu R, Shen Q, Xu P, et al. Phagocytosis of microglia in the central nervous system diseases［J］. Mol Neurobiol, 2014, 49: 1422-1434.
［7］Sahoo AK, Dandapat J, Dash UC, et al. Features and outcomes of drugs for combination therapy as multi-targets strategy to combat Alzheimer's disease ［J］. J Ethnopharmacol, 2018, 215: 42-73.
［8］潘娅岚, 马勇, 郭杨, 等. 脊髓康对脊髓损伤大鼠脊髓组织结构及神经生长因子表达的影响［J］. 中国实验方剂学杂志, 2014, 20(15): 144-149.
［9］Kluge MG, Kracht L, Abdolhoseini M, et al. Impaired microglia process dynamics post-stroke are specific to sites of secondary neurodegeneration ［J］. Glia, 2017, 65(12): 1885-1899.
［10］Nomura K, Vilalta A, Allendorf DH, et al. Activated Microglia desialylate and phagocytose cells via neuraminidase, galectin-3, and mer tyrosine kinase［J］. J Immunol, 2017, 198(12): 4792.
［11］Bellver-Landete V, Bretheau F, Mailhot B, et al. Microglia are an essential component of the neuroprotective scar that forms after spinal cord injury ［J］. Nat Commun, 2019, 10(1): 518.
［12］Bodhankar S, Lapato A, Chen Y, et al. Role for microglia in sex differences after ischemic stroke: importance of M2 ［J］. Metab Brain Dis, 2015, 30(6): 1515-1529.
［13］陈锴, 蔡梦溪, 晏梓钧, 等. 小胶质细胞M1/M2极化状态在脊髓损伤中作用的研究进展［J］. 脊柱外科杂志, 2019, 17(3): 216-219.
［14］Zeng WX, Han YL, Zhu GF, et al. Hypertonic saline attenuates expression of Notch signaling and proinflammatory mediators in activated microglia in experimentally induced cerebral ischemia and hypoxic BV-2 microglia［J］. Bmc Neuroscience, 2017, 18(1): 32.
［15］Sackmann V, Ansell A, Sackmann C, et al. Anti-inflammatory (M2) macrophage media reduce transmission of oligomeric amyloid beta in differentiated SH-SY5Y cells［J］. Neurobiol Aging, 2017, 60(4): 173-182.
［16］孙福荣. 脊髓康对大鼠脊髓损伤后轴突再生微环境的作用机制研究［D］. 南京中医药大学, 2013.
［17］Guo Y, Ma Y, Pan YL, et al. Jisuikang, a Chinese herbal formula, increases neurotrophic factor expression and promotes the recovery of neurological function after spinal cord injury ［J］. Neural Regen Res, 2017, 12(9): 1519-1528.
［18］郭杨, 马勇, 潘娅岚, 等. 脊髓康对大鼠脊髓损伤后神经功能恢复及脑源性神经营养因子表达的影响［J］. 中国康复理论与实践, 2014, 20(8): 701-708.
［19］Ma SF, Chen YJ, Zhang JX, et al. Adoptive transfer of M2 macrophages promotes locomotor recovery in adult rats after spinal cord injury［J］. Brain Behav Immun, 2015, 45(1): 157-170.
［20］张睿, 李冰洁, 李庆林. 酸枣仁皂苷A对脂多糖诱导小胶质细胞活化的影响及神经保护作用［J］. 中国临床药理学与治疗学, 2018, 23(2): 126-131.

[1]	王艳, 张金辉, 赵永辰, 刘宏祥, 刘亚维, 苗欢欢, 杨信才. 红杉黄酮通过下调PI3K/AKT信号通路抑制胃癌细胞增殖侵袭等干细胞特性[J]. 中国临床药理学与治疗学, 2023, 28(5): 508-513.
[2]	眭玉霞, 庄捷, 庄敏, 黄桂. Delicaflavone通过PI3K/AKT/mTOR通路调控上皮-间质转化抑制吉非替尼耐药肺癌PC-9/GR细胞迁移和侵袭[J]. 中国临床药理学与治疗学, 2022, 27(6): 614-621.
[3]	叶小弟, 缪云萍, 陈爱瑛, 程敏, 田雪君, 郑高利. 裸鼠原位非肌层浸润性膀胱癌模型的优选及验证[J]. 中国临床药理学与治疗学, 2022, 27(4): 473-480.
[4]	马旺博, 马月, 范方田, 张语涵, 常静雯, 周智华. 参芪扶正注射液干预低糖介导的免疫抑制微环境作用及其机制研究[J]. 中国临床药理学与治疗学, 2022, 27(2): 136-143.
[5]	张静, 高丹阳, 于康, 林晓婉, 曹颖, 刘晓, 缪慧慧, 李天佐. 不同吸入麻醉药对新生期大鼠神经胶质细胞活化的影响[J]. 中国临床药理学与治疗学, 2022, 27(12): 1391-1399.
[6]	李岩岩, 方骁杰, 尹鑫, 孙曦, 饶春晖. circZNF124通过靶向miR-4262调控结直肠癌SW620细胞增殖、迁移及侵袭的机制研究[J]. 中国临床药理学与治疗学, 2022, 27(11): 1231-1239.
[7]	潘蓉蓉, 支英豪, 金永喜, 周夏慧. 茶黄素调节CaMKK2/AMPK信号通路对脑出血大鼠神经元凋亡和血脑屏障的影响[J]. 中国临床药理学与治疗学, 2022, 27(11): 1240-1246.
[8]	吴林, 郝颖香, 张蕾, 张天雪, 高明晶, 冷玉芳. 帕立骨化醇通过HMGB1/TLR4/NF-κB信号通路减轻小鼠肠缺血再灌注损伤[J]. 中国临床药理学与治疗学, 2022, 27(1): 1-7.
[9]	江建波, 孙谦, 丁凤菲. 细胞外离子微环境调控睡眠觉醒[J]. 中国临床药理学与治疗学, 2021, 26(7): 794-801.
[10]	王叶情, 王宇红, 刘检, 赵洪庆, 邹蔓姝. 胆囊收缩素与抑郁症的相关性研究进展[J]. 中国临床药理学与治疗学, 2021, 26(6): 672-679.
[11]	江玉华, 支慧, 卢林明, 田大皓, 王晓庆, 葛钰, 谢尚富, 王琪. 皖南蝮蛇毒抑瘤组分-I逆转原代胃癌相关成纤维细胞促癌作用[J]. 中国临床药理学与治疗学, 2021, 26(4): 389-394.
[12]	陈林俊, 陈文斌. 岩大戟内酯B通过阻断PI3K/Akt/NF-κB通路抑制结肠癌细胞的增殖和转移[J]. 中国临床药理学与治疗学, 2021, 26(3): 250-257.
[13]	蒋伟宇, 胡旭栋, 陈云琳, 阮超越, 许楠健, 王扬, 徐顶立, 张佳铭, 李豪杰, 马维虎. 白藜芦醇抑制脊髓损伤兔的NLRP3炎症小体活化[J]. 中国临床药理学与治疗学, 2020, 25(8): 850-856.
[14]	黄康，李玲，叶啟发，彭贵主. 肝细胞肝癌中长链非编码RNA LINC00844的表达及对细胞增殖和迁移能力的抑制作用[J]. 中国临床药理学与治疗学, 2020, 25(4): 366-372.
[15]	沈成，杨年钊，王明海，吴立胜，朱志强. RNA甲基化酶NSUN2对胃癌细胞侵袭和迁移的影响及机制研究[J]. 中国临床药理学与治疗学, 2020, 25(4): 380-386.

脊髓康对大鼠脊髓损伤区小胶质细胞/神经元的影响

Effect of JiSuiKang on the microglia infiltration and neuron survival in the spinal cord injury area of rats

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价