中国病理生理杂志2024,Vol.40Issue(1) :118-125.DOI:10.3969/j.issn.1000-4718.2024.01.015

针刺足三里对CFA大鼠尾壳核中A1R/cAMP/p-CREB信号通路的影响

Effect of acupuncture on A1R/cAMP/p-CREB signaling pathway in cau-date putamen of CFA rats

张庆祥 周萌萌 霍明珠 常洪恩 司雨欣 张祐霖 房钰鑫
中国病理生理杂志2024,Vol.40Issue(1) :118-125.DOI:10.3969/j.issn.1000-4718.2024.01.015

针刺足三里对CFA大鼠尾壳核中A1R/cAMP/p-CREB信号通路的影响

Effect of acupuncture on A1R/cAMP/p-CREB signaling pathway in cau-date putamen of CFA rats

张庆祥 1周萌萌 1霍明珠 1常洪恩 1司雨欣 1张祐霖 1房钰鑫1
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作者信息

  • 1. 天津中医药大学针灸推拿学院,天津中医药大学实验针灸学研究中心,天津 301600
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摘要

目的:观察针刺对完全弗氏佐剂(CFA)大鼠尾壳核(CPu)中的腺苷A1受体(A1R)/环磷酸腺苷(cAMP)/cAMP反应元件结合蛋白(CREB)信号通路的影响,探讨针刺治疗炎性痛的潜在机制.方法:将64只6~8周龄雄性Wistar大鼠运用随机数字法随机分为盐水组、模型组(CFA组)、CFA+手针针刺(MA)组、CFA+溶剂二甲基亚砜(DMSO)组、CFA+A1R激动剂2-氯-N6-环戊基腺苷(CCPA)组、CFA+A1R拮抗剂8-环戊基-1,3-二丙基黄嘌呤(DPCPX)组、CFA+MA+DMSO组和CFA+MA+DPCPX组,每组8只.采用右侧足底皮内注射CFA制作关节炎炎性痛模型.针刺组于造模后第2天针刺大鼠双侧足三里穴,每次30 min,每天1次,共7 d.采用足底热辐射痛阈值评判大鼠疼痛反应;ELISA检测CPu脑区cAMP含量变化;Western blot检测CPu脑区PKA和CREB蛋白表达及磷酸化水平的变化;免疫荧光染色检测CPu脑区A1R表达情况.结果:与盐水组比较,CFA造模显著降低大鼠热痛阈值(P<0.01);与CFA组比较,CFA+MA组和CFA+CCPA组大鼠的热痛阈值均显著升高(P<0.05或P<0.01);与CFA+MA+ DMSO组比较,CFA+MA+DPCPX组大鼠的热痛阈值显著降低(P<0.05).与盐水组和CFA组比较,CFA+MA组大鼠CPu脑区中的A1R蛋白相对表达量和阳性细胞数均显著增加(P<0.05或P<0.01).与盐水组相比,CFA+MA组大鼠CPu脑区中的cAMP含量显著减少,p-CREB蛋白水平显著降低(P<0.05);与CFA+DMSO组相比,CFA+MA+ DMSO组和CFA+CCPA组的cAMP含量显著减少,p-CREB蛋白水平显著下降(P<0.01);与CFA+MA+DMSO组相比,CFA+MA+DPCPX组cAMP含量显著增加(P<0.01),p-PKA和p-CREB蛋白水平显著升高(P<0.05或P<0.01).结论:针刺双侧足三里可以缓解CFA大鼠的炎性疼痛,其机制可能与A1R/cAMP/p-CREB信号通路有关.

Abstract

AIM:To observe the effect of acupuncture on adenosine A1 receptor(A1R)in the caudate puta-men(CPu)of complete Freund's adjuvant(CFA)rats,and to explore the potential mechanism of acupuncture in treat-ment of inflammatory pain.METHODS:Sixty-four 6~8-week-old male Wistar rats were randomly divided into saline group,model group(CFA group),CFA+manual acupuncture(MA)group,CFA+solvent dimethyl sulfoxide(DMSO)group,CFA+A1R agonist 2-chloro-N6-cyclopentyladenosine(CCPA)group,CFA+A1R antagonist 8-cyclopentyl-1,3-di-propylxanthine(DPCPX)group,CFA+MA+DMSO group and CFA+MA+DPCPX group.In MA groups,on the 2nd day af-ter modeling,the rats were needled at Zusanli points on both sides,30 min at a time,once per day,for 7 d.Pain threshold of plantar thermal radiation was used to observe the pain response of the rats.The content of cyclic adenosine monophos-phate(cAMP)in the CPu was detected by ELISA.The protein expression and phosphorylation levels of protein kinase A(PKA)and cAMP response element-binding protein(CREB)were detected by Western blot.The expression of A1R in the CPu was detected by immunofluorescence staining.RESULTS:Compared with saline group,CFA modeling signifi-cantly lowered the thermal pain threshold of the rats(P<0.01).Compared with CFA group,the thermal pain threshold of the rats in CFA+MA group and CFA+CCPA group was significantly increased(P<0.05 or P<0.01).Compared with CFA+ MA+DMSO group,the thermal pain threshold of the rats in CFA+MA+DPCPX group was decreased(P<0.05).Compared with CFA group,A1R protein relative expression level and positive cells in the CPu of the rats in CFA+MA group were in-creased(P<0.05 or P<0.01).Compared with saline group,cAMP content and p-CREB protein level in the CPu of the rats in CFA+MA group were decreased(P<0.05).Compared with CFA+DMSO group,cAMP content and p-CREB pro-tein level in CFA+MA+DMSO and CFA+CCPA groups were significantly decreased(P<0.01).Compared with CFA+MA+ DMSO group,the levels of cAMP,p-PKA and p-CREB in CFA+MA+DPCPX group were significantly increased(P<0.05 or P<0.01).CONCLUSION:Acupuncture on bilateral Zusanli can relieve inflammatory pain in CFA rats,and its mech-anism may be related to A1R/cAMP/p-CREB signaling pathway.

关键词

针刺/炎性疼痛/腺苷A1受体/环磷酸腺苷/蛋白激酶A/cAMP反应元件结合蛋白

Key words

acupuncture/inflammatory pain/adenosine A1 receptor/cyclic adenosine monophosphate/pro-tein kinase A/cAMP response element-binding protein

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基金项目

国家自然科学基金资助项目(81973944)

天津市自然科学基金资助项目(22JCYBJC00990)

天津市研究生科研创新项目(2022SKY232)

天津中医药大学研究生科研创新项目(YJSKC-20221022)

出版年

2024
中国病理生理杂志
中国病理生理学会

中国病理生理杂志

CSTPCD北大核心
影响因子:1.065
ISSN:1000-4718
参考文献量7
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