鞘内注射人羊水外泌体对小鼠神经病理性痛的影响  

Effect of intrathecal exosomes derived from human amniotic fluid on neuropathic pain in mice

作　　者：张莎 许晶妹 李平[3] 钟涛[1] 王锷[1,2] 郭曲练[1] 杨勇[1,2] Zhang Sha;Xu Jingmei;Li Ping;Zhong Tao;Wang E;Guo Qulian;Yang Yong(Department of Anesthesiology,Xiangya Hospital,Central South University,Changsha 410008,China;National Clinical Research Center for Geriatric Disorders,Xiangya Hospital,Central South University,Changsha 410008,China;Department of Obstetrics,Xiangya Hospital,Central South University,Changsha 410008,China)

机构地区：[1]中南大学湘雅医院麻醉科,长沙410008 [2]中南大学湘雅医院国家老年疾病临床研究中心,长沙410008 [3]中南大学湘雅医院产科,长沙410008

出　　处：《中华麻醉学杂志》2023年第1期67-71,共5页Chinese Journal of Anesthesiology

基　　金：湖南省自然科学基金(2022JJ30975)。

摘　　要：目的评价鞘内注射人羊水外泌体对小鼠神经病理性痛的影响。方法清洁级健康雄性昆明小鼠18只,7~8周龄,体质量30~35 g,按照随机数字法分为3组(n=6):假手术组(Sham组)、坐骨神经选择性损伤组(SNI组)和坐骨神经选择性损伤+人羊水外泌体组(SNI+hAF exo组)。采用坐骨神经选择性损伤法制备小鼠神经病理性痛模型。另取3只小鼠,制备神经病理性痛模型;于造模后1、2和3 d时,鞘内注射PKH-26标记的人羊水外泌体7μl;给药结束后10 h处死小鼠,荧光显微镜下观察损伤侧腰膨大脊髓背角摄取人羊水外泌体情况。于造模后第1、2和3天,SNI+hAF exo组鞘内注射1μg/μl人羊水外泌体7μl,Sham组和SNI组鞘内注射PBS 7μl。分别于造模前1 d和造模后1、3、5、7 d时测定机械缩足反应阈(MPWT)。造模后7 d痛阈测定结束后处死小鼠取同侧脊髓腰膨大,采用Western blot法检测损伤侧脊髓CD11b、IL-1β和IL-10的表达。结果荧光显微镜下可见,神经病理性痛小鼠损伤侧腰膨大脊髓背角可以摄取羊水外泌体。与Sham组比较,SNI组造模后3~7 d时MPWT降低,脊髓CD11b和IL-1β表达上调(P<0.05),IL-10表达差异无统计学意义(P>0.05)。与SNI组比较,SNI+hAF exo组造模后3~5 d时MPWT升高,脊髓CD11b和IL-1β表达下调,IL-10表达上调(P<0.05)。结论鞘内注射人羊水外泌体可减轻小鼠神经病理性痛,其机制可能与介导小胶质细胞从M1型向M2型极化,减轻神经炎症有关。Objective To evaluate the effect of intrathecal exosomes derived from human amniotic fluid(hAF exo)on neuropathic pain induced by spared nerve injury(SNI)in mice.Methods Eighteen clean-grade healthy male Kunming mice,aged 7-8 weeks,weighing 30-35 g,were divided into 3 groups(n=6 each)using a random number table method:sham operation group(Sham group),SNI group,and SNI+hAF exo group.Spared nerve injury was produced by exposing the sciatic nerve and its branches and ligation and transection of tibial nerve and common fibular nerve in anesthetized mice.Another three mice were selected to develop the model of neuropathic pain after anesthesia.PKH-26 labeled hAF exo 7μl was intrathecally injected on days 1,2 and 3 after developing the model.The mice were sacrificed at 10 h after the end of administration,and the uptake of hAF exo by the dorsal horn of the injured lumbar enlargement of the spinal cord was observed with the fluorescence microscope.On 1,2 and 3 days after developing the model,1μg/μl hAF exo 7μl was intrathecally injected in SNI+hAF exo group,and PBS 7μl was intrathecally injected in Sham group and SNI group.The mechanical paw withdrawal threshold(MPWT)was measured at 1 day before and 1,3,5 and 7 days after operation.And then the mice were sacrificed after measurement of the pain threshold at 7 days after developing the model,and the ipsilateral lumbar enlargement of the spinal cord was taken for determination of the expression of CD11b,interleukin-1beta(IL-1β)and IL-10 by Western blot.Results The dorsal horn of the lumbar enlargement of the spinal cord on the injured side could absorb hAF exo with the fluorescence microscope.Compared with Sham group,the MPWT was significantly decreased at 3-7 days after developing the model,the expression of CD11b and IL-1βwas up-regulated(P<0.05),and no significant change was found in the expression of IL-10 in SNI group(P>0.05).Compared with SNI group,the MPWT was significantly increased at 3-7 days after developing the model,the expression of CD11b and IL-1βwa

关 键 词：外泌体 羊水 注射 脊髓 神经痛 

分 类 号：R614[医药卫生—麻醉学]