CCL2中和性抗体减轻皮肤/肌肉切口牵拉介导的大鼠慢性术后疼痛机制研究  

Mechanism of CCL2 neutralizing antibody in skin/muscle incision and retraction‑induced chronic postoperative pain in rats

作　　者：吴昊澎 曹明 陆建华 Wu Haopeng;Cao Ming;Lu Jianhua(Department of Anesthesiology,General Hospital of Southern Theatre Command of PLA,Guangzhou 510010,China)

机构地区：[1]中国人民解放军南部战区总医院麻醉科,广州510010

出　　处：《国际麻醉学与复苏杂志》2025年第2期139-145,共7页International Journal of Anesthesiology and Resuscitation

基　　金：南部战区总医院院内科技计划(2021NZC029)。

摘　　要：目的探究趋化因子配体2(CCL2)中和性抗体对皮肤/肌肉切口牵拉(SMIR)大鼠疼痛、行为影响及调控机制。方法192只雄性SD大鼠按照随机数字表法分为假手术组、SMIR组、SMIR+IgG组和SMIR+CCL2组,每组48只。假手术组预先鞘内注射10μl无菌磷酸盐缓冲液(PBS)液;SMIR组预先鞘内注射10μl无菌PBS溶液,2 h后行SMIR;SMIR+IgG组预先鞘内注射10μl IgG(10μg),2 h后行SMIR;SMIR+CCL2组大鼠预先鞘内注射10μl CCL2中和性抗体(10μg),2 h后行SMIR。4组大鼠分别于鞘内注射前3 d(T−3)、鞘内注射当天(T0)及鞘内注射后第1(T_(1))、3(T_(3))、5(T_(5))、7(T_(7))、14(T_(14))、21(T_(21))、28(T_(28))、35(T_(35))天测定造模足和对侧足疼痛行为学[缩足、抬腿次数,机械缩足阈值(MWT)及热缩足潜伏期(TWL)]。取造模后第7天各组大鼠血清和双侧背根神经节(DRG),采用酶联免疫吸附测定(ELISA)法测定血清肿瘤坏死因子⁃α(TNF⁃α)、白细胞介素(IL)⁃1β、IL⁃6水平,免疫印迹法(Western blot)检测DRG中CCL2、趋化因子C⁃C⁃基元受体2(CCR2)、谷氨酸受体2B(NR2B)蛋白水平和磷脂酰肌醇3激酶(PI3K)/蛋白激酶B(Akt)通路相关蛋白[PI3K、磷酸化PI3K(p⁃PI3K)、Akt、磷酸化Akt(p⁃Akt)、p⁃PI3K/PI3K、p⁃Akt/Akt]水平。结果与假手术组比较,SMIR组和SMIR+IgG组大鼠T_(1)、T_(3)、T_(5)、T_(7)、T_(14)、T_(21)、T_(28)、T_(35)时造模足MWT较低(均P<0.05);与SMIR组和SMIR+IgG组比较,SMIR+CCL2组大鼠T_(1)、T_(3)、T_(5)、T_(7)、T_(14)、T_(21)、T_(28)、T_(35)时造模足MWT较高(均P<0.05)。4组大鼠各时点对侧足MWT、造模足和对侧足TWL差异均无统计学意义(均P>0.05)。与假手术组比较,SMIR组和SMIR+IgG组大鼠T_(1)、T_(3)、T_(5)、T_(7)、T_(14)、T_(21)、T_(28)、T_(35)时造模足和对侧足缩足、抬腿次数较多(均P<0.05);与SMIR组和SMIR+IgG组比较,SMIR+CCL2组大鼠T_(1)、T_(3)、T_(5)、T_(7)、T_(14)、T_(21)、T_(28)、T_(35)时造模足和对侧足缩足、抬Objective To explore the effect of CC motif chemokine ligand 2(CCL2)neutralizing antibody on pain and behavior in rats with skin/muscle incision and retraction(SMIR)and its underlying regulatory mechanisms.Methods According to the random number table method,192 male SD rats were divided into four groups(n=48):a sham surgery group,a SMIR group,a SMIR+IgG group,and a SMIR+CCL2 group.The sham surgery group was intrathecally injected with 10μl sterile phosphatebuffered saline(PBS);the SMIR group was intrathecally injected with 10μl sterile PBS followed by SMIR 2 h later;the SMIR+IgG group was intrathecally injected with 10μl IgG(10μg)followed by SMIR 2 h later;and the SMIR+CCL2 group was intrathecally injected with 10μl CCL2 neutralizing antibody(10μg)followed by SMIR 2 h later.Their pain behaviors,including rat foot withdrawal,lifting times,mechanical withdrawal threshold(MWT),and thermal withdrawal latency(TWL)in the affected and contralateral paws were measured 3 days before intrathecal injection(T−3),on the day of injection(T0),and at postinjection days 1(T_(1)),3(T_(3)),5(T_(5)),7(T_(7)),14(T_(14)),21(T_(21)),28(T_(28)),and 35(T_(35)).On postmodeling day 7,rats were sacrificed to collect serum and bilateral dorsal root ganglia(DRG).The levels of serum tumor necrosis factorα(TNFα),interleukin(IL)1β,and IL6 were measured by enzymelinked immunosorbent assay(ELISA).The levels of CCL2,CC motif chemokine receptor 2(CCR2),glutamate ionotropic receptor NmethylDaspartate type subunit 2B(NR2B),and proteins associated with the phosphatidylinositol 3kinase(PI3K)/protein kinase B(Akt)signaling pathway,including PI3K,phosphoPI3K(pPI3K),Akt,phosphoAkt(pAkt),pPI3K/PI3K,and pAkt/Akt in the DRG were detected by Western blot.Results Compared with the sham surgery group,the SMIR and SMIR+IgG groups exhibited significant decreases in MWT in the affected paw at T_(1),T_(3),T_(5),T_(7),T_(14),T_(21),T_(28),and T_(35)(all P<0.05).Compared with the SMIR and SMIR+IgG groups,the SMIR+CCL2 group had significantly increases in MW

关 键 词：趋化因子配体2中和性抗体 皮肤/肌肉切口牵拉 慢性术后疼痛 炎症 

分 类 号：R614[医药卫生—麻醉学]