| 孙海榉,王思思,项璇儿,等.电针对痛觉敏化诱发大鼠脊髓背角p38丝裂原活化蛋白激酶/肿瘤坏死因子-α的影响[J].浙江中医药大学学报,2019,43(12):1301-1309. |
| 电针对痛觉敏化诱发大鼠脊髓背角p38丝裂原活化蛋白激酶/肿瘤坏死因子-α的影响 |
| Effect of Electroacupuncture on Pain Conversion and Content of P38 Mitogen-Activated Protein Kinase and Tumor Necrosis Factor-α in Spinal Dorsal Horn in Hyperalgesia Priming Rats |
| DOI：10.16466/j.issn1005-5509.2019.12.001 |
| 中文关键词: 电针 痛觉敏化诱发 足三里 昆仑 脊髓 p38 MAPK TNF-α |
| 英文关键词: electroacupuncture hyperalgesic priming Zusanli Kunlun spinal p38 MAPK TNF-α |
| 基金项目:浙江省基础公益研究计划(LQ18H270001)；国家自然科学基金青年项目(81603692)；国家自然科学基金面上项目(81473772)；浙江省医药卫生科技计划项目(2016KYA154)；2018年浙江省大学生科技创新活动暨新苗人才计划资助项目(2018R410055)；浙江省一流学科(中医学)资助(浙政办函[2017]) |
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| 中文摘要: |
| [目的]观察电针(electroacupuncture，EA)对痛觉敏化诱发模型大鼠造模侧热痛阈(thermal paw-withdrawal latency，TPWL)、机械性痛阈(mechanical paw-withdrawal threshold，MPWT)及脊髓背角p38丝裂原活化蛋白激酶(mitogen-activated protein kinase，MAPK)和肿瘤坏死因子-α(tumor necrosis factor-α，TNF-α)表达的影响。[方法]第一部分:按完全随机法将健康雄性SD大鼠分为3组，其中空白组5只、假敏化组5只、敏化组6只用于MPWT检测；各组其余6只大鼠分别用于用于TPWL检测。造模方法:敏化组采用1%角叉菜胶100μL左后足足底皮下注射，待痛阈恢复至基础水平后，以100ng·25μL-1前列腺素E2(prostaglandin E2，PGE2)25μL注射于左足足背中央，建立痛转化模型；空白组两次均注射等量0.9%氯化钠注射液；假敏化组第1次注射等量0.9%氯化钠注射液，第2次注射等量PGE2，浓度与剂量同敏化组。大鼠造模前、第1次注射后4、24、48、72h和7d，第2次注射(第1次注射后8d)后1、4、24、48h检测MTWP和TPWL。以Western blot法检测第2次注射后48h造模侧脊髓背角p38 MAPK和TNF-α蛋白表达。第二部分:将大鼠随机分为假敏化组、敏化组、EA组、假电针(sham electroacupuncture，sham EA)组，每组12只，其中6只用于MPWT检测；其余6只用于TPWL检测。假敏化组和敏化组造模方法同第一部分，EA组和sham EA组造模方法同敏化组。第1次注射后，EA组大鼠于双侧“足三里”“昆仑”穴行EA干预，1次/d，共10次；sham EA组仅针刺入大鼠皮下。各组大鼠痛阈检测时间同第一部分。以Western blot法检测第2次注射后48h造模侧脊髓背角p38 MAPK和TNF-α蛋白表达。[结果]与空白组和假敏化组比较，第1次注射后4、24、48和72h，第2次注射后4、24、48h，敏化组造模侧MPWT和TPWL显著降低(P＜0.01，P＜0.01)，说明痛觉敏化诱发模型造模成功。与空白组和假敏化组比较，敏化组大鼠造模侧脊髓背角p38 MAPK和TNF-α表达增高(P＜0.01，P＜0.01)。与敏化组比较，第1次注射后24、48、72h及第2次注射后4、24、48h，EA组造模侧的MPWT和TPWL明显升高(P＜0.01，P＜0.01)，而sham EA组并不能提高大鼠痛阈，痛阈变化趋势与敏化组一致。与假敏化组比较，敏化组造模侧脊髓背角中p38 MAPK和TNF-α表达增高(P＜0.01，P＜0.01)。与敏化组比较，EA组造模侧脊髓背角p38 MAPK和TNF-α表达减低(P＜0.05，P＜0.05)，而sham EA组p38 MAPK和TNF-α表达高于假敏化组(P＜0.01，P＜0.01)，与敏化组无统计学差异(P＞0.05，P＞0.05)。[结论]EA具有良好的镇痛作用并能够干预痛转化效应，其机制可能与下调造模侧脊髓背角p38 MAPK和TNF-α的表达有关。 |
| 英文摘要: |
| [Objective]To observe the effect of electroacupuncture(EA) on thermal paw-withdrawal latency(TPWL) and mechanical paw-withdrawal threshold (MPWT) and its regulation of p38 mitogen-activated protein kinase(MAPK) and tumor necrosis factor-α(TNF-α) in spinal dorsal horn in hyperalgesia priming rats, so as to explore its spinal mechanism underlying improvement of inflammatory pain. [Methods]In the first part, thirty-four SD rats were randomized into blank group, sham hyperalgesic priming group and hyperalgesic priming group, 5 rats in blank group, 5 rats in sham hyperalgesic priming group and 6 in hyperalgesic priming group for the test of MPWTs, other 6 rats in each group were used for the test of TPWLs. Hyperalgesic priming model was established by subcutaneous injection of 1% carrageenan into the same left hind paw(the first injection),followed by injection of 100ng·25μL-1 prostaglandin E2(PGE2) 25μL (the second injection) into the dorsum pedis of the same hind paw 8 days after the first injection. The ipsilateral paw MPWT and TPWL was detected before and 4,24,48,72h and 7 days after the first injection, 1,4,24 and 48h after the second injection. In the second part，forty- eight SD rats were randomly divided into sham hyperalgesic priming group, hyperalgesic priming group, EA group and sham electroacupuncture(sham EA) group, with 12 rats in each group. Six rats in each group were used for the test of MPWTs and the other six rats were used for the test of TPWLs. The sham hyperalgesic priming and hyperalgesic priming models were established in the same way as the first part. Both“Zusanli(ST 36) and“Kunlun”(BL 60)were punctured with filiform needles in sham EA group and the acupuncture points were also stimulated with EA, 2Hz/100Hz, 0.5～1.5mA, increased 0.5mA per 10min, for 30mins in EA group，once per day, until the end of experiment to test the effect of EA on rat's lateral MPWT. Protein content in ipsilateral spinal dorsal horn was assayed by Western blot 48h after the second injection. [Results]In the first part, compared with blank group and sham hyperalgesic priming group, the MPWT and TPWL at 4,24,48 and 72h after the first injection and 4,24,48h after the second injection were significantly decreased in hyperalgesic priming group(P＜0.01, P＜0.01).The protein expression of p38 MAPK and TNF-α in spinal dorsal horn was significantly higher in hyperalgesic priming group than in blank group and sham hyperalgesic priming group(P＜0.01,P＜0.01).In the second part, compared with hyperalgesic priming group, the MPWT and TPWL at 24, 48 and 72h after the first injection and 4, 24 and 48h after the second injection increased significantly in EA group(P＜0.01, P＜0.01).The protein expression of p38 MAPK and TNF-α in spinal dorsal horn was significantly higher in hyperalgesic priming group than in sham hyperalgesic priming group(P＜0.01, P＜0.01), but considerably lower in EA group than in hyperalgesic priming group(P＜0.05, P＜0.05).The protein expression of p38 MAPK and TNF-α was significantly higher in sham EA group than in sham hyperalgesic priming group(P＜0.01, P＜0.01) and there was no statistically significant difference between sham EA group and hyperalgesic priming group(P＞0.05, P＞0.05). [Conclusion] EA has good effect on pain conversion, and its mechanism may be related to the effect in down-regulating p38 MAPK and TNF-α in spinal dorsal horn. |
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