背根神经节P2X3受体表达上调在糖尿病神经痛中的作用  

Role of upregulated P2X3 expression in dorsal root ganglia during diabetic neuropathic pain

作　　者：陈卢杭 费雪瑜 康玉蓉 王涵芝 瞿思颖 李想 何晓芬 方剑乔[1,2,3] 蒋永亮 CHEN Luhang;FEI Xueyu;KANG Yurong;WANG Hanzhi;QU Siying;LI Xiang;HE Xiaofen;FANG Jianqiao;JIANG Yongliang(Third Clinical Medical College and Rehabilitation Medical College of Zhejiang Chinese Medical University,Hangzhou 310053;China.2.Key Laboratory of Acupuncture and Neurology of Zhejiang Province,Hangzhou 310053;.3.Institute of Acupuncture and Moxibustion,Zhejiang University of Traditional Chinese Medicine,Hangzhou 310053)

机构地区：[1]浙江中医药大学第三临床医学院康复医学院,杭州310053 [2]浙江省针灸神经病学研究重点实验室,杭州310053 [3]浙江中医药大学针灸研究所,杭州310053

出　　处：《中国实验动物学报》2021年第5期593-599,共7页Acta Laboratorium Animalis Scientia Sinica

基　　金：国家自然科学基金(81774389,81804181);国家级大学生创新创业训练计划(202010344015,202010344017);浙江省大学生科技创新活动计划暨新苗人才计划(2020R410012)。

摘　　要：目的本研究拟观察注射链脲佐菌素(streptozotocin,STZ)后大鼠不同时期背根神经节(dorsal root ganglion,DRG)上嘌呤受体(purinergic receptor subtype,P2X3)的表达情况及P2X3受体拮抗剂TNP-ATP对糖尿病神经痛(diabetic neuropathic pain,DNP)大鼠的干预作用。方法(1)20只健康雄性SD大鼠随机选取6只选取作为正常组,其余14只大鼠予以腹腔注射STZ,剔除未成模的2只大鼠,分别观察造模前(Base),造模后7 d、14 d、21 d的机械缩爪阈值(paw withdrawal threshold,PWT)变化情况;并在上述各时间点取大鼠L4-L6 DRG,采用免疫荧光法检测L4-L6 DRG上P2X3阳性细胞表达情况。(2)将25只健康雄性SD大鼠随机选取6只作为正常+生理盐水(Control+NS)组,其余19只予以STZ注射,剔除未成模大鼠1只,成功建立DNP的大鼠随机分为模型+生理盐水(DNP+NS)组,模型+50 nmol P2X3抑制剂TNP-ATP组(DNP+50 nmol TNP-ATP),模型+100 nmol P2X3抑制剂TNP-ATP组(DNP+100 nmol TNP-ATP),每组6只;STZ注射14 d后,DNP+TNP-ATP组分别按照上述剂量予以足背注射TNP-ATP溶液,其余两组分别予以注射等量NS,观察注射后0.5、1、1.5 h大鼠PWT变化。同时观察连续注射7 d药物对大鼠PWT的影响。结果(1)与正常组比较,模型组大鼠第7天、第14天及第21天空腹血糖显著升高;与正常组比较,模型组大鼠Day 7 PWT无明显改变,第14天、第21天PWT显著降低。免疫荧光结果显示,与正常组相比,STZ注射7、14、21 d后,DNP大鼠L4、L5 DRG上P2X3的阳性细胞的表达显著升高,STZ注射14、21 d后,DNP大鼠L6 DRG上P2X3的阳性细胞的表达显著升高。(2)TNP-ATP干预前,DNP+NS组与DNP+50 nmol TNP-ATP组、DNP+100 nmol TNP-ATP组PWT无显著差异;干预0.5 h后,与DNP+NS组、DNP+50 nmol TNP-ATP组相比,DNP+100 nmol TNP-ATP组PWT明显升高,效果持续至1 h。(3)连续注射7 d TNP-ATP后,DNP+100 nmol TNP-ATP组PWT较DNP+NS组与DNP+50 nmol TNPATP组明显升高。结论腹腔注射STZ可成功建立DNP大鼠模型,背根神经节P2X3表Objective To observe the expression of purinergic receptor subtype P2X3(P2X3)in dorsal root ganglion(DRG)of rats at different stages after injection of streptozotocin(STZ),and the intervention effect of P2X3 receptor antagonist TNP-ATP in diabetic neuralgia(DNP)rats.Methods(1)Six of 20 healthy male SD rats were randomly selected as the normal group,and the other 14 rats were intraperitoneally injected with STZ.Two STZ treated rats without modeling were excluded.The paw withdrawal threshold(PWT)was observed before and 7(Day 7),14(Day 14)and 21 days(Day 21)after modeling.The L4-L6 DRG of rats were collected at the above time points,and the expression of P2X3 positive cells was detected by immunofluorescence.(2)Six of 25 healthy male SD rats were randomly selected as the normal+normal saline(Control+NS)group,and the remaining 19 rats were injected with STZ.One STZ injected rat was excluded.The rats with successfully established DNP were randomly divided into a model+normal saline(DNP+NS)group,model+50 nmol P2X3 inhibitor TNP-ATP(DNP+50 nmol TNP-ATP)group,and model+100 nmol P2X3 inhibitor TNP-ATP(DNP+100 nmol TNP-ATP)group,with six rats in each group.Fourteen days after STZ injection,the DNP+TNP-ATP group was injected with 100 nmol TNP-ATP solution in the dorsum of the foot,whereas the other two groups were injected with the same volume of NS,and the PWT was observed at 0.5,1 and 1.5 h after injection.The effect of drug injection on PWT was also observed after 7 days.Results(1)Compared with the normal group,fasting blood glucose was significantly increased in rats of the model group on Day 7,Day 14 and Day 21.Compared with the normal group,there was no significant change in the PWT of the model group at Day 7,but there was a significant decrease in PWT at Day 14 and Day 21.Immunofluorescence showed that the expression of P2X3 positive cells on L4 and L5 DRG from the DNP group was significantly increased 7,14 and 21 days after STZ injection compared with the normal group.(2)Before TNP-ATP intervention,there were no si

关 键 词：糖尿病神经痛 背根神经节 P2X3受体 

分 类 号：Q95-33[生物学—动物学]