从MAPKs信号通路探讨健脾通络方的消肿镇痛作用机制  

Anti-swelling and Analgesic Mechanism of Jianpi Tongluo Prescription from MAPKs Signaling Pathway

作　　者：明瑞蕊 方罗昌婷 王潇潇 胡智星 杨超 徐腾腾 王宏洁[1] 刘春芳[1] 林娜[1] MING Ruirui;FANG-LUO Changting;WANG Xiaoxiao;HU Zhixing;YANG Chao;XU Tengteng;WANG Hongjie;LIU Chunfang;LIN Na(Institute of Chinese Materia Medica,China Academy of Chinese Medicine Sciences,Beijing 100700,China)

机构地区：[1]中国中医科学院中药研究所,北京100700

出　　处：《中国实验方剂学杂志》2023年第12期85-93,共9页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：中国中医科学院科技创新工程重大攻关项目(C12021A03808);国家“重大新药创制”科技重大专项(2019ZX09731-002)。

摘　　要：目的:观察健脾通络方的消肿和镇痛作用,并初步探索其作用机制。方法:120只ICR小鼠,分为正常组、模型组、健脾通络方低、中、高剂量组(5、10、20 g·kg^(-1))、阳性药塞来昔布组(0.03 g·kg^(-1)),每组10只,每天1次经口灌服,采用小鼠完全弗氏佐剂(CFA)诱导慢性炎性痛模型、二甲苯诱导耳肿胀实验、热板实验和乙酸扭体实验4种急、慢性模型观察不同剂量的健脾通络方的消肿和镇痛作用;采用酶联免疫吸附测定法(ELISA)检测慢性炎性痛小鼠血清和致炎足中前列腺素E_(2)(PGE_(2))、白细胞介素(IL)-1β、IL-6、肿瘤坏死因子-α(TNF-α)的含量表达,蛋白免疫印迹法(Western blot)检测慢性炎性痛小鼠致炎足中水通道蛋白(AQP)1、AQP3、环氧化酶(COX)1、COX2、丝裂原活化蛋白激酶(MAPKs)家族的蛋白表达情况,探索健脾通络方的初步作用机制。结果:与正常组比较,二甲苯所致模型小鼠耳肿胀度显著升高,热板实验小鼠热缩足潜伏期缩短(P<0.01);与模型组比较,健脾通络方能显著提高二甲苯所致小鼠耳肿胀度抑制率(P<0.05,P<0.01),延长小鼠乙酸致扭体潜伏期和减少扭体反应次数(P<0.05,P<0.01)。与正常组比较,慢性炎性痛小鼠足肿胀度显著升高,机械痛阈值降低同时冷敏痛阈值升高(P<0.05,P<0.01),致炎足中的AQP1和AQP3的蛋白含量增加,血清和(或)致炎足中IL-1β、IL-6、TNF-α、PGE_(2)及致炎足COX2含量升高,致炎足p-p38 MAPK、p-JNK、p-ERK的蛋白表达水平升高(P<0.01);与模型组比较,健脾通络方显著降低慢性炎性痛小鼠的足肿胀度、升高机械痛阈值并降低冷敏痛阈值,镇痛持续4 h,且给药后2 h为镇痛最佳时间点(P<0.05,P<0.01);进一步,健脾通络方下调慢性炎性痛小鼠致炎足中AQP1和AQP3的蛋白含量,降低血清和(或)致炎足中IL-1β、IL-6、TNF-α、PGE_(2)及致炎足COX2含量,但对COX1无明显影响,同时降低致炎足p-p38 MAPK、p-JNK、p-ERK的蛋白表Objective:To observe the anti-swelling and analgesic effects of Jianpi Tongluo prescription(JPTL) and to explore its mechanism initially.Method:A total of 120 ICR mice were divided into normal group,model group,JPTL low-,medium-and high-dose groups (5,10,20 g·kg^(-1)) and positive drug(celecoxib,0.03 g·kg^(-1)) group,with 10 in each group (po,once a day).Complete freund's adjuvant (CFA) was used to induce the model of chronic inflammatory pain,and xylene-induced ear swelling test,hot plate test and acetic acid writhing test were performed to observe the anti-swelling and analgesic effects of different doses of JPTL in these four acute and chronic models.Further,enzyme-linked immunosorbent assay (ELISA) was used to detect the expressions of prostaglandin E_(2) (PGE_(2)),interleukin-1β (IL-1β),interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in serum and inflammatory paw of mice with chronic inflammatory pain,and the expressions of aquaporin 1 (AQP1),aquaporin 3 (AQP3),cyclooxygenase 1 (COX1),cyclooxygenase 2(COX2) and mitogen-activated protein kinases (MAPKs) in inflammatory paw were detected by Western blot,to explore the preliminary mechanism of JPTL.Result:Compared with the conditions in the normal group,there was a significant increase in the ear swelling of xylene-induced model mice,a shortened paw withdrawal latency in the hot plate test (P<0.01).Compared with the model group,JPTL remarkably increased the inhibition rate of xylene-induced ear swelling (P<0.05,P<0.01),prolonged the latency period of writhing caused by acetic acid and reduced the number of writhing (P<0.05,P<0.01).Compared with normal group,the degree of feet swelling in chronic inflammatory pain mice was significantly increased,the threshold of mechanical pain was decreased and the threshold of cold pain was increased (P<0.05,P<0.01),the protein contents of AQP1 and AQP3 in inflammatory feet were increased,and the contents of IL-1β,IL-6,TNF-α,PGE_(2) and COX2 in inflammatory feet were increased in serum and/or inflammatory fee

关 键 词：健脾通络方 消肿 镇痛 抗炎 水通道蛋白 炎症介质 丝裂原活化蛋白激酶 

分 类 号：R2-0[医药卫生—中医学] R33R289R684.3