【摘要】：背景:已經(jīng)證實針灸對腰背痛、膝關節(jié)痛等急性疼痛等有較好療效,但是對于慢性疼痛的效果存在爭議。目的:建立新西蘭兔慢性關節(jié)炎疼痛模型,探討針灸治療慢性關節(jié)炎疼痛機制。方法:(1)32只新西蘭兔使用4%木瓜蛋白酶0.3 mL注射雙后肢膝關節(jié)腔內,制成兔膝骨性關節(jié)炎模型。隨機分為4組,即生理鹽水+電針組,生理鹽水+假電針組,k-阿片受體拮抗劑(nor-Binaltorphimine,nor-BNI)+電針組,nor-BNI+假電針組,每組8只。nor-BNI用量1 mg/kg,1次/d,連續(xù)3 d;電針干預時間為實驗動物給予藥物后2 h,1次/d,30 min/次,直至實驗動物被處死當天結束;假電針組取穴、針刺方法及時間同電針組,但不通電刺激。使用BBB評分對各組進行下肢行為學評分。給藥1,3,5,7日分別處死,分離脊髓組織后多聚甲醛固定,使用免疫熒光法檢測脊髓組織白細胞介素17及白細胞介素17受體A、NR1的表達水平;(2)另取24只新西蘭兔,12只兔建立膝骨性關節(jié)炎模型后隨機分為模型+生理鹽水組、模型+2μg白細胞介素17抗血清組;剩余12只分為對照+生理鹽水組、對照+2μg白細胞介素17抗血清組。以2μg白細胞介素17抗血清溶于10μL生理鹽水中進行椎管內給藥,1次/d,連續(xù)3 d。使用BBB評分對新西蘭兔下肢功能進行行為測試,每次行為測試后,Western-blot測定樣品中p-NR1,白細胞介素17R的表達水平。結果與結論:(1)nor-BNI+電針組BBB評分明顯增高(P0.05);而脊髓組織白細胞介素17及白細胞介素17受體A、NR1的表達水平明顯減低(P0.05);(2)使用nor-BNI拮抗阿片類受體后,nor-BNI+電針組脊髓組織NR1的表達水平與nor-BNI+假電針組相比無明顯差異(P0.05);(3)使用白細胞介素17抗血清處理后,模型+2μg白細胞介素17抗血清組BBB評分明顯增高(P0.05);而該組脊髓組織白細胞介素17及NR1的表達水平明顯減低(P0.05),但仍高于2個對照組(P0.05);(4)實驗證實,慢性關節(jié)炎疼痛原因為白細胞介素17可以使脊髓組織NR1表達增加從而引起;電針刺激可以明顯改善關節(jié)炎模型新西蘭兔的下肢疼痛,這種作用是由于電針刺激減少脊髓組織白細胞介素17而非調節(jié)白細胞介素17受體A發(fā)揮作用。
[Abstract]:Background: acupuncture and moxibustion have been proved to have good effect on acute pain, such as low back pain, knee pain and so on, but the effect of chronic pain is controversial. Objective: to establish a pain model of chronic arthritis in New Zealand rabbits and to explore the mechanism of acupuncture and moxibustion in the treatment of chronic arthritis pain. Methods: (1) 32 New Zealand rabbits were injected with 0.3 mL of papain 0.3 mL into the knee joint of both hind limbs to establish a rabbit knee osteoarthritis model. They were randomly divided into 4 groups: normal saline electroacupuncture group, nor-BNI pseudoEA group. The dosage of nor-BNI was 1 mg / kg / d in each group for 3 consecutive days; the time of electroacupuncture intervention was 30 min/ after 2 h of administration of the drug, until the end of the day on which the animals were killed; the acupuncture method and time of the sham electroacupuncture group were the same as those of the electroacupuncture group. But no electrical stimulation. BBB score was used to evaluate the behavior of lower extremities in each group. 5 and 7 days after administration, the spinal cord tissue was isolated and fixed with paraformaldehyde. The expression of interleukin-17 (IL-17) and interleukin-17 receptor (A1-NR1) in spinal cord tissue were detected by immunofluorescence assay. (2) A total of 24 New Zealand rabbits and 12 rabbits were randomly divided into normal saline group after the establishment of knee osteoarthritis model. Model 2 渭 g interleukin 17 antiserum group, the remaining 12 groups were divided into control saline group and control 2 渭 g interleukin 17 antiserum group. 2 渭 g interleukin 17 antiserum was dissolved in 10 渭 L normal saline for 3 days. The lower limb function of New Zealand rabbits was tested with BBB score. After each behavioral test, the expression of p-NR1 and IL-17R in the samples was determined by Western-blot. Results and conclusion: (1) the BBB score of nor-BNI electroacupuncture group was significantly increased (P0.05), while the expression of interleukin-17 and interleukin-17 receptor Agnor-1 in spinal cord tissue was significantly decreased (P0.05); (2). Nor-BNI antagonized opioid receptor was used to antagonize NR1 in spinal cord tissue of nor-BNI electroacupuncture group (P0.05). There was no significant difference between the nor-BNI pseudoacupuncture group and the control group (P0.05); (3) after treatment with interleukin-17 antiserum. The BBB score of model 2 渭 g interleukin 17 antiserum group was significantly increased (P0.05), while the expression of interleukin 17 and NR1 in spinal cord tissue was significantly decreased (P0.05), but still higher than that in two control groups (P0.05); (4). The cause of chronic arthritis pain is that interleukin-17 can increase NR1 expression in spinal cord and electroacupuncture can significantly improve lower limb pain in rabbits with arthritis. This effect is due to the role of electroacupuncture stimulation in reducing interleukin-17 (IL-17) in spinal cord rather than regulating IL-17 receptor A (IL-17 receptor A).
【作者單位】： 西南醫(yī)科大學附屬醫(yī)院康復醫(yī)學科;西南醫(yī)科大學附屬醫(yī)院骨與關節(jié)外科;
【基金】：四川省中醫(yī)藥管理局基金項目(2014-K-125) 西南醫(yī)科大學青年基金項目(2014QN-107) 四川省衛(wèi)計委普及應用項目(17PJ176)~~
【分類號】：R245;R-332

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