电针对坐骨神经分支选择性损伤大鼠脊髓L-Arg/NO/cGMP通路的影响  

Electroacupuncture relieved neuropathic pain by suppressing L-Arg/NO/cGMP pathway in the lumbar spinal cord in rats with spared nerve injury

作　　者：闫丽萍[1] 侯保权 李守栋[1] 王玲玲[1] 马骋[1] YAN Li⁃ping;HOU Bao⁃quan;LI Shou⁃dong;WANG Ling⁃ling;MA Cheng(Jiangsu Province Key Laboratory of Acupunc⁃ture⁃moxibustion,Nanjing University of Chinese Medicine,Nanjing 210029,China;Jiangsu Keygen Biotech,LTD.,Nanjing 210012)

机构地区：[1]南京中医药大学江苏省针灸学重点实验室,南京210029 [2]江苏凯基生物技术股份有限公司,南京210012

出　　处：《针刺研究》2019年第12期893-897,905,共6页Acupuncture Research

基　　金：国家自然科学基金面上项目(No.81173329);江苏省自然科学基金基础研究计划(No.BK2008458)

摘　　要：目的:观察坐骨神经分支选择性损伤(SNI)大鼠脊髓相应节段左旋精氨酸(L-Arg)转运体2(CAT-2)、一氧化氮合酶(NOS)与一氧化氮(NO)和环鸟苷酸(cGMP)的变化及电针对其的影响,探讨电针干预神经病理性痛的脊髓机制。方法:SD大鼠随机分为假手术组、模型组、电针组和NOS抑制剂(LNAME)组,每组30只。采用SNI法制备神经病理性痛模型,假手术组仅分离不结扎。电针组电针大鼠损伤侧"委中""环跳"穴,频率2 Hz,波宽0.6 ms,起始电流强度1 mA,每10 min递增1 mA,刺激时间30 min,1次/d,连续7 d;L-NAME组予L-NAME 60 mg·kg^-1·d^-1腹腔注射,连续7 d。造模前及SNI后10、16 d分别测定机械痛阈。RT-PCR法测定脊髓CAT-2 mRNA与诱导型NOS(iNOS)mRNA的表达,Western blot法测定脊髓iNOS的表达,硝酸/亚硝酸还原酶法与放射免疫分析法分别测定NO(NO2-/NO3-)与cGMP的含量。结果:与假手术组比较,SNI后10 d其他各组机械痛阈值均降低(P<0.01);与模型组比较,电针组和LNAME组SNI后16 d时机械痛阈值均升高(P<0.01,P<0.05)。与假手术组比较,模型组脊髓CAT-2mRNA、iNOS mRNA、iNOS蛋白表达及NO2-/NO3-与cGMP含量均升高(P<0.01,P<0.05);与模型组比较,电针组和L-NAME组脊髓CAT-2 mRNA、iNOS mRNA、iNOS蛋白表达及NO2-/NO3-与cGMP含量均下降(P<0.05,P<0.01),且L-NAME组CAT-2 mRNA、iNOS mRNA、iNOS蛋白表达比电针组降低更显著(P<0.05)。结论:电针缓解疼痛的作用机制之一,可能是通过有效下调神经病理性痛状态下脊髓L-Arg/NO/cGMP通路的功能而实现的。Objective To observe the effect of electroacupuncture(EA)on changes of expression of L-Arg transporter 2(CAT-2)m RNA and nitric oxide synthase(i NOS)m RNA and protein and contents of NO and c GMP of L4-L6 segments of spinal cord in rats with spared nerve injury(SNI),so as to reveal its mechanism underlying reducing neuropathic pain.MethodsA total of 120 male SD rats were randomly divided into sham operation,model,EA and NOS inhibitor(N omega-Nitro-L-arginine methyl ester hydrochloride,L-NAME)groups,with 30 rats in each group.The neuropathic pain model was established by ligating and cutting the tibial nerve and the common peroneal nerve.EA(2 Hz,1-3 m A)was applied to"Weizhong"(BL40)and"Huantiao"(GB30)on the damaged hindlimb for 30 min,once daily from day 11 to 17 after SNI.Rats of the L-NAME group received i.p.of L-NAME(60 mg·kg^-1·d^-1)for 7 consecutive days.The mechanical pain threshold(PT)was determined before and 10 and 16 d after SNI,respectively.The expression levels of CAT-2 m RNA and i NOS m RNA,and i NOS protein in the L4-L6 segments of the spinal cord were detected by using reverse transcription-polymerase chain reaction(RT-PCR)and Western blot,respectively,and the contents of NO and cGMP of L4-L6 assayed using nitrate/nitrite reductase method and radioimmunoassay,respectively.Results After modeling,the PT was significantly decreased on day 10 and 16 after SNI in comparison with the sham operation group and their own baseline data of pre-operation in each group(P<0.01),and remarkably increased in the EA and L-NAME groups relevant to the model group on day 16(P<0.01,P<0.05).Compared with the sham operation group,the expression levels of CAT-2 mRNA and i NOS m RNA and protein,as well as the contents of NO2-/NO3-and c GMP were significantly up-regulated in the model group(P<0.05,P<0.01).Following EA intervention,the levels of CAT-2 m RNA and i NOS m RNA and i NOS protein,and NO2-/NO3-and cGMP contents were all reversed in both EA and L-NAME groups(P<0.05,P<0.01).The effect of EA was significantly superio

关 键 词：电针 神经病理性痛 脊髓 左旋精氨酸转运体 一氧化氮合酶 一氧化氮 环鸟苷酸 

分 类 号：R245.97[医药卫生—针灸推拿学]