蛇床子素通过抑制髓核致炎大鼠脊髓背角CXCL1/CXCR2的表达发挥抗炎镇痛作用  

Effect of osthole on CXCL1/CXCR2 of rat model with inflammatory radicular pain induced by nucleus pulposus in spinal horn

作　　者：高凤娇 杨琳 张嘉明 魏明[1] 贺秋兰[1] 邹学农[4,5] 孙来保 GAO Feng-jiao;YANG Lin;ZHANG Jia-ming;WEI Ming;HE Qiu-lan;ZOU Xue-nong;SUN Lai-bao(Dept of Anesthesiology,the First Affiliated Hospital of Sun Yat-sen University,Guangzhou 510080,China;Dept of Anesthesiology,Guangzhou Puyu Central Hospital,Guangzhou 511400,China;Dept of Anesthesiology,the Sixth Affiliated Hospital of Sun Yat-sen University,Guangzhou 510655,China;Dept of Spine Surgery,the First Affiliated Hospital of Sun Yat-sen University,Guangzhou 510080;Guangdong Provincial Key Laboratory of Orthopedics and Traumatology,Guangzhou 510080,China)

机构地区：[1]中山大学附属第一医院麻醉科,广东广州510080 [2]广州市番禺中心医院麻醉科,广东广州511400 [3]中山大学附属第六医院麻醉科,广东广州510655 [4]中山大学附属第一医院脊柱外科,广东广州510080 [5]广东省骨科学重点实验室,广东广州510080

出　　处：《中国药理学通报》2020年第3期347-354,共8页Chinese Pharmacological Bulletin

基　　金：广东省骨科学重点实验室基金(No 2016B030301002);番禺区科技计划项目(NO2018Z0439)。

摘　　要：目的研究蛇床子素(osthole,Ost)对大鼠髓核源性神经根痛的抗炎镇痛作用。方法雄性SD大鼠60只随机分为3组:髓核致炎组(NP组,n=36)、Sham组(n=12)、Blank组(n=12),检测术前和术后3、7、14、21、28 d的50%MWT;q-PCR检测术后各时间点脊髓背角CXCL1、CXCR2和TNF-α的mRNA水平,Western blot检测术后各时间点脊髓背角CXCL1/CXCR2的蛋白水平。另外雄性SD大鼠64只髓核致炎后随机均分为4组:生理盐水组(NP+NS),DMSO组(NP+DMSO),Ost组(NP+Ost),CXCL1中和抗体组(NP+AF-515),于术后d 3经硬膜外注射相应药物50μL。给药前后多个时点均检测50%MWT。于术后d 7取L5脊髓背角,通过Western blot检测各组脊髓背角CXCL1/CXCR2的蛋白水平。结果NP组术后各时间点的50%MWT较Sham组均降低(P<0.05)。NP组术后各时间点脊髓背角CX-CL1、CXCR2和TNF-α的mRNA水平均较Sham组升高(P<0.05),且NP组术后各时间点脊髓背角CXCL1/CXCR2的蛋白水平较Sham组升高(P<0.05)。NP+Ost组给药后50%MWT比给药前升高(P<0.05),NP+AF515组在给药后3 h 50%MWT升高,持续到给药后12 h(P<0.05)。NP+Ost组术后7d脊髓背角CXCL1/CXCR2蛋白表达降低(P<0.05)。结论脊髓背角CXCL1/CXCR2可能参与了大鼠髓核源性神经根痛的发生发展,蛇床子素通过抑制脊髓背角CXCL1/CXCR2的表达而缓解髓核致炎大鼠神经根性疼痛。Aim To investigate the effects of osthole on anti-inflammation and analgesia on nucleus pulposus-derived radiculopathy in rats and the underlying mech-anism.Methods Sixty male SPF SD rats were ran-domly assigned to three groups:NP group(n=36,nu-cleus pulposus-derived neuropathic pain model was es-tablished by placing around 0.4 mg of the rat autolo-gous nucleus pulposus in the left L5 dorsal root gangli-on),sham group(n=12,rat nucleus pulposus was re-moved but not placed in the L5 dorsal root ganglion,with other procedures remained same as those in the NP group.),and blank group(n=12,no surgery was performed)in first part.50%mechanical withdrawal threshold(50%MWT)was measured using Von Frey filaments before and 3,7,14,21,and 28 days after op-eration.L5 spinal dorsal horns were taken out from rats in sham group 7 days postoperatively and in NP group 3,7,14,21,and 28 days postoperatively,of which the expression levels of CXCL1 mRNA,CXCR2 mRNA and TNF-αmRNA were evaluated with q-PCR and protein expression levels of CXCL1 and CXCR2 determined by Western blot.Sixty-four male SPF SD rats were ran-domly assigned to four groups(n=16 in each group):NP+NS group(NP+physiological saline),NP+DM-SO group(NP+dimethylsulfoxide),NP+Ost group(NP+osthole),and NP+AF-515 group(NP+CX-CL1 neutralizing antibody)in second part.All four groups were epidurally injected with corresponding drugs.50%MWT was measured at the various time points before and after operation.L5 spinal dorsal horn was taken out 7 days after operation and protein expres-sion levels of CXCL1 and CXCR2 were determined with Western blot.Results Significant decreases of 50%MWT(P<0.05)were observed in NP group at all postoperative time points,which were also signifi-cantly lower than those in sham group at the corre-sponding postoperative time points(P<0.05),sugges-ting a successful modelling of nucleus pulposus-derived neuropathic pain.The expression levels of CXCL1 mR-NA,CXCR2 mRNA and TNF-αmRNA as well as the protein expression levels of CXCL1 and CXCR2 protein in L5

关 键 词：蛇床子素 CXCL1 CXCR2 脊髓背角 炎性神经根痛 髓核 

分 类 号：R-332[医药卫生] R284.1