Zhang Xiao-Hong, Li Fei-Fei, Qi Yue, Ming Cai-Rong, Li Ying, Pan Si-Teng, Liu Si-Jia, Ma Tie-Ming
Graduate School, Liaoning University of Traditional Chinese Medicine, Shenyang 110032, China.
State Key Laboratory of Natural and Biomimetic Drugs of Peking University, Beijing 100191.
Zhen Ci Yan Jiu. 2020 Apr 25;45(4):299-304. doi: 10.13702/j.1000-0607.190263.
To observe the effect of electroacupuncture (EA) on degranulation of intraperitoneal mast cells (MCs) and expression of mitogen-activated protein kinase (MAPK) signaling related proteins, tumor necrosis factor-α(TNF-α) and interleukin-6 (IL-6) in urticaria rats, so as to reveal its mechanisms underlying improvement of urticaria.
Thirty-two SD rats were randomly divided into control，model，EA and medication groups (＝8 in each group). The urticaria model was established by using passive cutaneous anaphylaxis (PCA) reaction method. EA (2 Hz /15 Hz, 1 mA) was applied to bilateral "Zusanli"(ST36), "Quchi "(LI11) and "Xuehai"(SP10) for 20 min，once daily for 7 consecutive days before antigen attack. Rats of the medication group were treated by gavage of Loratadine(1 mg•kg•d)for 7 days. The diameter of cutaneous Evan's blue spots was measured to evaluate the severity of PCA. Intraperitoneal fluid smears were prepared to observe the degranulation state of MCs. The contents of TNF-α and IL-6 in the intraperitoneal fluid were detected by ELISA, and the expression of extracellular signal-regulated kinase (ERK), phosphorylated (p)-ERK, c-Jun N-terminal kinase (JNK), p-JNK, P38MAPK and p-P38MAPK of the acquired intraperitoneal MCs was detected by Western blot.
The diameter of cutaneous Evan's blue spot was significantly increased in the model group than that in the control group (<0.01), and considerably decreased in both EA and medication groups compared with the model group(<0.01). After modeling，the percentage of degranulated MCs, contents of TNF-α and IL-6, and expression levels of ERK, p-ERK, JNK, p-JNK, P38MAPK and p-P38MAPK were remarkably increased in the mo-del group than those in the control group (<0.01, <0.05). After the treatment, the percentage of degranulated MCs, contents of TNF-α and IL-6, and expression levels of p-ERK, JNK, p-JNK and p-P38MAPK were obviously decreased in both EA and medication groups relevant to the model group (<0.01, <0.05), while no significant changes were found in the expression of ERK in both EA and medication groups, and P38MAPK in the EA group. Compared with the model and EA groups, expression levels of P38MAPK were down-regulated in the medication group (<0.05).
EA can reduce skin allergic reaction in rats with urticaria, which may be related to its effects in inhibiting the degranulation of intraperitoneal MCs, down-regulating the expression of MAPK signaling-related proteins and the level of pro-inflammatory factors TNF-α and IL-6 in intraperitoneal MCs.
观察电针（EA）对荨麻疹大鼠腹腔肥大细胞（MCs）脱颗粒及丝裂原活化蛋白激酶（MAPK）信号相关蛋白、肿瘤坏死因子-α（TNF-α）和白细胞介素-6（IL-6）表达的影响，以揭示其改善荨麻疹的作用机制。
将32只SD大鼠随机分为对照组、模型组、电针组和药物组（每组8只）。采用被动皮肤过敏反应（PCA）法建立荨麻疹模型。在抗原攻击前，电针（2Hz/15Hz，1mA）双侧“足三里”（ST36）、“曲池”（LI11）和“血海”（SP10）20分钟，每天1次，连续7天。药物组大鼠灌胃氯雷他定（1mg•kg•d）7天。测量皮肤伊文思蓝斑直径以评估PCA的严重程度。制备腹腔液涂片观察MCs的脱颗粒状态。采用酶联免疫吸附测定（ELISA）法检测腹腔液中TNF-α和IL-6的含量，采用蛋白质印迹法检测获取的腹腔MCs中细胞外信号调节激酶（ERK）、磷酸化（p）-ERK、c-Jun氨基末端激酶（JNK）、p-JNK、P38丝裂原活化蛋白激酶（P38MAPK）和磷酸化（p）-P38MAPK的表达。
模型组皮肤伊文思蓝斑直径显著大于对照组（P<0.01），电针组和药物组均显著小于模型组（P<0.01）。造模后，模型组MCs脱颗粒百分比、TNF-α和IL-6含量以及ERK、p-ERK、JNK、p-JNK、P38MAPK和p-P38MAPK的表达水平均显著高于对照组（P<0.01，P<0.05）。治疗后，电针组和药物组MCs脱颗粒百分比、TNF-α和IL-6含量以及p-ERK、JNK、p-JNK和p-P38MAPK的表达水平均显著低于模型组（P<0.01，P<0.05），而电针组和药物组ERK的表达以及电针组P38MAPK的表达无明显变化。与模型组和电针组相比，药物组P38MAPK的表达水平下调（P<0.05）。
电针可减轻荨麻疹大鼠的皮肤过敏反应，这可能与其抑制腹腔MCs脱颗粒、下调MAPK信号相关蛋白的表达以及腹腔MCs中促炎因子TNF-α和IL-6的水平有关。
