摘要
目的观察星形胶质细胞和小胶质细胞标志物在1型糖尿病外周神经病变小鼠脊髓中的表达变化。方法SPF级健康雄性C57BL/6小鼠30只,6周龄,体重18~22 g,采用随机数字表法分为两组:对照组(C组)和糖尿病组(DM组)。DM组采用链脲佐菌素(STZ)(100 mg/kg,连续2 d腹腔注射)制备1型糖尿病小鼠模型,C组给予同等剂量柠檬酸钠缓冲液连续2 d。记录两组小鼠造模前、造模后1、2、4、6、8、10周的体重、随机血糖、机械缩足反应阈(MWT)、热缩足潜伏期(TWL)。生化检测分别选取C组和DM组1周、10周的脊髓。采用Western blot法测定L 4-L 6脊髓脂肪酸结合蛋白7(FABP7)、星形胶质细胞特异性标记物胶质纤维酸性蛋白(GFAP)、小胶质细胞标记物(CD11b、iba1)表达量,Elisa法测定小鼠脊髓组织中肿瘤坏死因子α(TNF-α)、IL-10浓度,免疫组化法测定足底表皮神经纤维密度(IENFD)。结果与C组比较,DM组造模后2、4、6、8、10周体重、MWT明显降低(P<0.05或P<0.01),造模后1、2、4、6、8、10周血糖明显升高(P<0.01),造模后4、6、8、10周TWL明显延长(P<0.05或P<0.01);造模后1周FABP7和GFAP表达量明显升高(P<0.05);造模后10周FABP7、CD11b、iba1表达量明显升高(P<0.05);造模后10周脊髓组织中IL-10、TNF-α浓度明显升高(P<0.05);造模后10周IENFD明显降低(P<0.05)。结论糖尿病模型小鼠造模后1周脊髓中星形胶质细胞活化但小胶质细胞无明显改变,而造模后10周星形胶质细胞已恢复正常但小胶质细胞活化,1型糖尿病外周神经病变中星型胶质细胞较小胶质细胞更早的激活。
Objective To observe the expression of astrocyte and microglia biomarkers in the spinal cord of mice with type 1 diabetic peripheral neuropathy.Methods Thirty SPF-grade healthy male C57BL/6 mice,6 weeks old,weighing 18-22 g,were divided into 2 groups by random number table method:control group(group C)and diabetes mellitus group(group DM).Mice in group DM was treated with streptozotocin(STZ)(100 mg/kg,intraperitoneally injected for 2 consecutive days)to make type 1 diabetic mice model,and the mice in group C were given the same dose of sodium citrate buffer for 2 consecutive days.The weight,blood glucose,mechanical withdrawal threshold(MWT)and thermal withdrawal latency(TWL)of mice in both groups were recorded before modeling and 1,2,4,6,8,and 10 weeks after modeling.The L 4-6 spinal cords of mice in group C and group DM 1 week and 10 weeks after modeling were taken for biochemical detection.The expressions of fatty acid-binding protein7(FABP7),astrocyte biomarker glial fibrillary acidic protein(GFAP),and microglia biomarker(CD11b and iba1)were detected by Western blot method.The levels of TNF-αand IL-10 were detected by enzyme-linked immunosorbent assay.Plantar intraepidermal nerve fiber density(IENFD)was detected by immunohistochemical method.Results Compared with group C,the weight and MWT in group DM significantly decreased 2,4,6,8,10 weeks after modeling(P<0.05 or P<0.01),the blood glucose significantly increased of group DM 1,2,4,6,8,10 weeks after modeling(P<0.01),and the TWL of group DM obviously extended 4,6,8,10 weeks after modeling(P<0.05 or P<0.01).The expressions of FABP7 and GFAP were elevated(P<0.05)1 week after modeling,the expressions of FABP7,CD11b,iba1 and the concentration of TNF-αand IL-10 within spinal cord tissues were significantly elevated in 10 weeks(P<0.05).However,the IENFD was reduced in 10 weeks after modeling.Conclusion The astrocytes in the spinal cord of the diabetic mice were active but the microglia did not change significantly 1 week after the modeling,and the astrocytes had returned to normal but the microglia were active in 10 weeks after the modeling.The outcomes indicate that in type 1 diabetic peripheral neuropathy,astrocytes are active earlier than microglia.
作者
简娇敏
纪健
陈冲
娄晓丽
朱涛
JIAN Jiaomin;JI Jian;CHEN Chong;LOU Xiaoli;ZHU Tao(Department of Anesthesiology Shanghai Songjiang Clinical Medical College of Nanjing Medical University,Shanghai 201600,China)
出处
《临床麻醉学杂志》
CAS
CSCD
北大核心
2020年第11期1111-1115,共5页
Journal of Clinical Anesthesiology
基金
江苏省研究生科研与实践创新计划项目(KYCX18_1503)。
