基于嘌呤配体P2X门控离子通道型受体7的资生肾气丸镇痛机制研究

Zisheng Shenqi Pills Relieve Pain via Purinergic Receptor P2X,Ligand-gated Ion Channel 7

目的:探索资生肾气丸对醋酸致小鼠扭体模型的镇痛作用。方法:将42只小鼠随机分为7组,分别给予相应药液灌胃,末次灌胃1 h后造模,建立醋酸致小鼠扭体模型。观察小鼠扭体次数,计算其扭体抑制率,应用酶联免疫吸附试验(ELISA)检测各组小鼠血清白细胞介素-1β(IL-1β)、前列腺素E 2(PGE 2)、神经生长因子(NGF)含量,采用实时PCR(RT-PCR)检测嘌呤配体P2X门控离子通道型受体7(P2X7R)mRNA、Nod样受体蛋白3(NLRP3)mRNA、NIMA相关激酶7(NEK7)mRNA的表达情况。结果:与模型组比较,随着时间的增加,资生肾气丸低剂量、中剂量、高剂量均可抑制小鼠扭体反应,降低扭体次数,增加小鼠扭体抑制率,降低小鼠血清中IL-1β、PGE_(2)、NGF的含量,降低P2X7RmRNA、NLRP3mRNA、NEK7mRNA的表达以抑制疼痛反应,以高剂量效果最优(P<0.05),与吲哚美辛、痛风舒片疗效相近。结论:资生肾气丸高剂量对小鼠血清疼痛因子的降低效果显著,其机制可能与嘌呤配体P2X门控离子通道型受体7信号通路有关。

Objective:To explore the pain-relieving effect of Zisheng Shenqi Pills on the mouse model of writhing induced by acetic acid.Methods:Forty-two mice were randomized into 7 groups and administrated with corresponding drug solutions by gavage.The model was established 1 h after the last administration.The writhing times of mice were observed,and the writhing inhibition rate was calculated.The enzyme-linked immunosorbent assay(ELISA)was employed to measure the levels of interleukin-1β(IL-1β),prostaglandin E 2(PGE 2),and nerve growth factor(NGF)in the serum.The mRNA levels of purinergic receptor P2X,ligand-gated ion channel 7(P2X7R),Nod-like receptor protein 3(NLRP3),and NIMA-related kinase 7(NEK7)were determined by RT-PCR.Results:Compared with the model group,Zisheng Shenqi Pills at low,medium,and high doses inhibited the writhing response,reduced the writhing times,increased the inhibition rate of writhing,and lowered the levels of IL-1β,PGE_(2),and NGF in the serum of mice.Moreover,they down-regulated the mRNA levels of P2X7R,NLRP3,and NEK7 to inhibit the pain response.The high dose group demonstrated the best effect(P<0.05),which was similar to that of indometacin and Tongfengshu Tablets.Conclusion:Zisheng Shenqi Pills at high doses can significantly lower the levels of pain factors in the serum of mice by regulating the P2X7R signaling pathway.