摘要
目的观察针刺对实验性自身免疫性脑脊髓炎(encephalomyelitis,EAE)小鼠神经系统损伤症状的改善程度和脊髓磷酸化丝裂原活化蛋白激酶(phosphorylated P38 mitogen-activated protease,p-p38MAPK)及血清IL-1β的影响,初步探究针刺对多发性硬化(multiple sclerosis,MS)实验动物模型-EAE小鼠的治疗作用和机制。方法将40只C57BL/6小鼠随机分为空白组、模型组、模型+针刺组(针刺组)、模型+激素组(激素组),每组10只。除空白组外,其余各组小鼠采用髓鞘少突胶质细胞糖蛋白33-55(myelin oligodendrocyte glycoprotein 33-55,MOG 33-55 )与完全弗氏佐剂诱导建立EAE模型。各组自造模第14天开始,每日上午同一时间干预治疗。空白组和模型组每日抓取、固定,并用生理盐水[0.1 mL/(10 g·d)]灌胃;针刺组针刺小鼠“大椎”、双侧“足三里”“肾俞”穴15分钟;激素组用醋酸泼尼松龙(0.6 mg/mL)灌胃[0.1 mL/(10 g·d)]治疗;以上各组均连续干预治疗15天。从造模当天开始,隔日测量体质量和神经功能评分评价小鼠行为学变化;于免疫第28天对各组小鼠进行眼球采血并留取脊髓组织,采用LFB染色技术检测小鼠脊髓脱髓鞘损伤程度;分别采用酶联免疫吸附法和蛋白质免疫印迹法检测小鼠血清IL-1β的含量及脊髓组织p-p38MAPK蛋白表达。结果针刺组和激素组小鼠体质量明显增加,神经功能评分明显降低,与模型组比较差异有统计学意义( P <0.05);与模型组相比,针刺组及激素组小鼠脊髓脱髓鞘程度明显减轻;免疫第28天时,模型组小鼠血清IL-1β表达与空白组相比水平升高( P <0.05);针刺组与激素组小鼠血清IL-1β水平与模型组相比均降低( P <0.05);模型组小鼠脊髓p-p38MAPK蛋白表达水平比空白组表达水平升高( P <0.05);而针刺组与激素组小鼠脊髓p-p38MAPK蛋白水平与模型组相比均降低( P <0.05)。结论针刺可以明显改善EAE小鼠神经系统损伤症状,降低EAE小鼠的神经功能评分;针刺缓解EAE模型小鼠神经系统损伤症状的机制可能与其能抑制EAE小鼠脊髓p38MAPK蛋白激活及其能降低小鼠血清IL-1β的表达水平有关。
Objective To observe the improvement degree of acupuncture on nervous system injury symptoms of experimental autoimmune encephalomyelitis (EAE) mice and its effects on spinal phosphorylated P38 mitogen-activated protease (p-p38 MAPK) and serum IL-1β, and to preliminarily explore the therapeutic effect and mechanism of acupuncture on multiple sclerosis (MS) animal model - EAE mice. Methods A total of 40 C57BL/6 mice were randomly divided into 4 groups: blank group, model group, model+acupuncture group (acupuncture group) and model+hormone group (hormone group), with 10 mice in each group. Except for the blank group, mice in other groups were immunized with emulsifier prepared by mixing myelin oligodendrocyte glycoprotein 33-55 (MOG 33-55 ) with complete Freund's adjuvant to establish EAE model. Starting from the 14th day of modeling, each group received intervention treatment at the same time every morning. The blank group and the model group were grasped and fixed daily, and were intragastrically administered with normal saline [0.1 mL/(10 g·d)]. In the acupuncture group,“Dazhui”(GV14;DU14) point, bilateral “Zusanli”(S36;ST36) points and bilateral “Shenshu”(B23;BL23) points were taken for routine disinfection and acupuncture intervention, each time for 15 min. The hormone group was intragastrically administered with [0.1 mL/(10 g·d)] of prednisolone acetate solution (0.6 mg/mL). All groups were treated continuously for 15 days. From the day of modeling, the behavioral changes of mice were closely observed, and the body weight of mice in each group was measured every other day and the neurological function was scored. On the 28 th day of immunization, blood was collected from the eyeballs of mice in each group and spinal cord tissue was collected. LFB staining was used to detect the degree of demyelination in spinal cord of mice. The expression of p-p38 MAPK protein in the spinal cord tissue of mice was detected by Western blot, and the content of IL-1β in serum of mice in each group was detected by enzyme-linked immunosorbent assay. Results The body weight of the mice in acupuncture group and hormone group significantly increased, and the neurological function score significantly decreased, comparing with the model group the difference was statistically significant ( P <0.05 ). Compared with model group, the degree of demyelination of spinal cord in acupuncture group and hormone group significantly reduced. On the 28 th day of immunization, the level of IL-1β expression in the model group was higher than that in the blank group ( P <0.05). The level of serum IL-1β in acupuncture group and hormone group was lower than that in model group ( P <0.05). The expression level of phosphorylated p38 MAPK protein p-p38 MAPK in the spinal cord of the model group was higher than that in the blank group ( P <0.05). However, the level of p-p38MAPK in spinal cord in acupuncture group and hormone group was lower than that in model group ( P <0.05). Conclusion Acupuncture can significantly improve the symptoms of nervous system injury in EAE mice and reduce the neurological function score of EAE mice. The mechanism of acupuncture alleviating the symptoms of nervous system injury in EAE model mice may be related to the fact that acupuncture can inhibit the activation of p38 MAPK protein in spinal cord of EAE mice and reduce the expression level of IL-1β in serum of mice.
作者
赵东杰
李琪
王玉华
董莎
常磊
张鹏
宋方亚
辛随成
ZHAO Dongjie;LI Qi;WANG Yuhua(School of Acupuncture-Moxibustion and Tuina, Beijing University of Chinese Medicine, Beijing 100029, China)
出处
《环球中医药》
CAS
2019年第4期486-492,共7页
Global Traditional Chinese Medicine
