腺苷A1受体介导艾灸镇痛作用研究

发布时间：2018-08-04 09:08

【摘要】：目的：明确腺苷A1受体是否参与艾灸的镇痛作用。方法：以C57BL/6J小鼠为研究对象,筛选基础热痛阈值在10 s-16 s的小鼠,随机分为空白组、造模组；造模组予小鼠右足底注射完全弗氏佐剂20μL建立炎症痛模型,造模后第4天,在小鼠右侧“足三里”处分别运用CCPA(A1受体激动剂)、CPT(A1受体拮抗剂)进行干预；模灸组小鼠于“足三里”穴位温和灸,施灸15 min；镇痛效应测定分别在造模前、造模后第4天(即艾灸治疗前)、艾灸后15min、45 min、75 min、105 min、135 min用PL-200热刺痛仪测定热痛阈；采用实时荧光定量PCR技术检测下丘脑AiR mRNA表达；统计分析采用SPSS23.0统计软件处理。结果：(1)炎症痛模型小鼠艾灸镇痛效应与空白组比较,模型组小鼠痛阈降低,差异有统计学意义(P0.05)；与模型组比较,模灸组小鼠痛阈显著提高,在15～75 min时差异有统计学意义(P0.05)。(2)腺苷A1受体对艾灸镇痛作用的影响1)腺苷A1受体激动剂对艾灸镇痛作用的影响①单独注射三种不同浓度激动剂(CCPA) (0.05 mM、0.1mM、0.2 mM)对模型组进行干预后,痛阈均升高；CFA+CCPA 0.05 mM组在注射后45～135 min时,较注射前升高(P0.05)；CFA+CCPA 0.1 mM 及 CFA+CCPA 0.2 mM组在注射后15～135 min时,均保持较高水平,与注射前比较有差异(P0.05)。②艾灸加三种不同浓度激动剂(CCPA) (0.05 mM、0.1mM、0.2 mM)对模灸组进行干预后,痛阈均升高；CFA+MOXI+CCPA 0.05 mM组在干预后,在15～135 min与模灸组比较,差异无统计学意义(P＞0.05);CFA+MOXI+CCPA0.1 mM组和CFA+MOXI+CCPA 0.2 mM组在干预后15～135 min,均比模灸组痛阈有所提高(P0.05)；与单独注射三种不同浓度CCPA(0.05 mM,0.1mM、0.2 mM)组比较,CFA+MOXI+CCPA 0.05 mM组痛阈值在15～75 min均有所提高(P0.05)；CFA+MOXI+CCPA 0.05 mM组在15 min, CFA+MOXI+CCPA 0.2 mM组在45min时痛阈均提高(P0.05)2)腺苷A1受体拮抗剂对艾灸镇痛作用的影响①单独注射三种不同浓度拮抗剂(CPT)(75 nM、150 nM、300 nM)对模型组进行干预,CFA+CPT 75 nM组及CFA+CPT 150 nM组在注射后15～105 min,CFA+CPT 300 nM组在注射后15～135 min时痛阈下降,与注射前比较有差异(P0.05);②艾灸加三种不同浓度拮抗剂(CPT)(75nM、50 nM、300 nM)对模灸组进行干预后,其痛阈均有所下降；与模灸组比较,CFA+MOXI+CPT 75 nM组、CFA+MOXI+CPT 150 nM组、CFA+MOXI+CPT 300 nM组在干预后15-105 min,痛阈均有所降低,差异有统计学意义(P0.05)；与单独注射三种不同浓度CPT(75 nM、150 nM、300 nM)组比较,CFA+MOXI+CPT 75 nM组、CFA+MOXI+CPT 150nM组在干预后15～75 min,痛阈均升高(P0.05); CFA+MOXI+CPT 300 nM组在干预后15～105 min,痛阈升高,差异有统计学意义(P0.05)。(3)下丘脑A1R mRNA表达的变化结果：与空白组比较,模型组下丘脑A1R mRNA表达下调2.23倍。与模型组比较,模灸组下丘脑A1R mRNA表达上调4.26倍。结论：1.艾灸“足三里”能提高炎症痛小鼠痛阂,镇痛效应在灸后约45 min达到高峰；2.局部腺苷A1受体参与艾灸镇痛；①激活穴位局部A1受体增强艾灸镇痛效应；②阻断穴位局部A1受体减弱艾灸镇痛效应；3.下丘脑腺苷A1受体可能参与艾灸镇痛。
[Abstract]:Objective: to determine whether adenosine A1 receptor is involved in the analgesic effect of moxibustion. Methods: taking C57BL/6J mice as the research object, the mice with basic heat pain threshold in 10 s-16 s were selected and randomly divided into blank group and model group. The model group was injected into the right foot of the mouse to establish an inflammatory pain model with completely Freund's adjuvant 20 L, and fourth days after making the model, on the right side of mice. " CCPA (A1 receptor agonist) and CPT (A1 receptor antagonist) were used respectively in Zusanli; moxibustion group mice were treated with mild moxibustion at "Zusanli" point and 15 min with moxibustion. The analgesic effect was measured before modeling, fourth days after moxibustion (before moxibustion treatment), 15min, 45 min, 75 min, 105 min, 135 min with PL-200 tantalgia. The AiR mRNA expression in the hypothalamus was detected by real time fluorescence quantitative PCR, and the statistical analysis was processed by SPSS23.0 statistical software. Results: (1) the analgesic effect of Moxibustion in the model mice was compared with that of the blank group. The pain threshold of the model group was lower, the difference was statistically significant (P0.05); compared with the model group, the pain threshold of moxibustion group was obvious. The difference was statistically significant at 15~75 min (P0.05). (2) the effect of adenosine A1 receptor on moxibustion analgesia 1) the effect of adenosine A1 receptor agonist on the analgesic effect of Moxibustion (1) alone injection of three different concentration agonists (0.05 mM, 0.1mM, 0.2 mM) to the model group after the dry prognosis, the pain threshold increased; CFA+CCPA 0.05 mM group At 45~135 min after injection, higher than before injection (P0.05), CFA+CCPA 0.1 mM and CFA+CCPA 0.2 mM in 15~135 min after injection, all maintained a higher level, compared with before injection (P0.05). 2. Moxibustion plus three different concentration agonists (CCPA) (0.05 mM, 0.1mM, 0.2 mM) to the moxibustion group after the prognosis, pain threshold increased. There was no significant difference between the XI+CCPA 0.05 mM group and the moxibustion group in the 15~135 min group (P > 0.05), and the CFA+MOXI+CCPA0.1 mM group and the CFA+MOXI+CCPA 0.2 mM group were 15~135 min compared with the moxibustion group (P0.05), and compared with the group of three different concentrations CCPA (0.05 mM, 0.2). The pain threshold in the 0.05 mM group was improved at 15~75 min (P0.05), the CFA+MOXI+CCPA 0.05 mM group was 15 min, the CFA+MOXI+CCPA 0.2 mM group increased the pain threshold at 45min (P0.05) 2) the effect of adenosine A1 receptor antagonist on the moxibustion analgesic effect (1) alone injection of three different concentrations of antagonists (150, 150, 300) to the model group. The pain threshold of A+CPT 75 nM group and CFA+CPT 150 nM group was 15~105 min after injection, and the pain threshold of CFA+CPT 300 nM group decreased at 15~135 min after injection (P0.05). 2. Moxibustion plus three different concentration antagonists (CPT) (75nM, 50 nM, 300) had the prognosis of moxibustion group, and the pain threshold decreased; compared with moxibustion group, The T 75 nM group, CFA+MOXI+CPT 150 nM group and CFA+MOXI+CPT 300 nM group were 15-105 min in the dry prognosis, and the pain threshold decreased. The difference was statistically significant (P0.05). Compared with the group of three different concentrations of CPT (75 nM, 150 nM, 300 nM) group, the 75 groups were 15~75 of the prognosis, and the pain threshold increased. The XI+CPT 300 nM group had a 15~105 min prognosis and a higher pain threshold (P0.05). (3) the changes in the expression of A1R mRNA in the hypothalamus: compared with the blank group, the expression of A1R mRNA in the hypothalamus was down 2.23 times. Compared with the model group, the expression of A1R mRNA expression in the hypothalamus of moxibustion group was up 4.26 times. Conclusion: 1. Moxibustion "Zusanli" can be raised. The pain gap of high inflammatory pain mice, the analgesic effect reached the peak about 45 min after moxibustion; 2. local adenosine receptor was involved in moxibustion analgesia; (1) activating the local A1 receptor to enhance the analgesic effect of moxibustion; (2) blocking the local A1 receptor to weaken the analgesic effect of moxibustion; 3. the adenosine A1 receptor in the hypothalamus could participate in moxibustion analgesia.
【学位授予单位】：成都中医药大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R245.81

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