摘要
目的:探讨胃饥饿素(Ghrelin)在糖尿病神经病理性疼痛中的作用及对大鼠脊髓嘌呤受体2X-4/NOD样受体-3(P2X4/NLRP3)表达的影响。方法:健康成年雄性SD大鼠40只,随机分为5组,每组8只:正常+Ghrelin组(NG组)、正常+[D-Lys3]-GHRP-6组(ND组)、糖尿病组(D组)、糖尿病+Ghrelin组(DG组)、糖尿病+Ghrelin+[D-Lys3]-GHRP-6组(DGD组)。[D-Lys3]-GHRP-6为Ghrelin特异性抑制剂。NG组腹腔注射Ghrelin 200μg/kg 6周,ND组腹腔注射[D-Lys3]-GHRP-6 50 mg/kg 6周。腹腔注射1%链脲佐菌素-柠檬酸盐缓冲液60 mg/kg制备大鼠糖尿病模型,造模成功后3 d,腹腔注射Ghrelin 200μg/kg 6周,DGD组腹腔注射Ghrelin 200μg/kg+[D-Lys3]-GHRP-6 50 mg/kg 6周。分别于第2,4,6周测定大鼠机械痛阈(MWT)及坐骨神经导速率(MNCV);6周后处死大鼠,尼氏染色观察脊髓病理学变化,电镜观察腓肠神经病理学改变,Western blot法检测脊髓Ghrelin、P2X4、NLRP3和IL-1β表达水平。结果:与NG组比较,ND组脊髓Nissl染色和腓肠神经电镜显示细胞结构正常,MWT和MNCV无明显变化,P2X4、NLRP3、IL-1β表达水平差异无统计学意义(P>0.05);D组脊髓Nissl染色和腓肠神经电镜显示细胞部分受损,MWT、MNCV显著降低;Ghrelin表达降低,P2X4、NLRP3、IL-1β显著升高(P<0.05)。与D组比较,DG组中脊髓Nissl染色和腓肠神经电镜显示细胞结构相对正常,MWT和MNCV明显升高,Ghrelin表达升高,P2X4、NLRP3、IL-1β表达降低(P<0.05)。与DG组比较,DGD组中脊髓Nissl染色和腓肠神经电镜细胞结构破坏严重,MWT和MNCV明显降低,Ghrelin表达降低,P2X4、NLRP3、IL-1β表达升高(P<0.05)。结论:Ghrelin可能通过P2X4/NLRP3通路参与了糖尿病神经病理性疼痛的调控。
Objective:To evaluate the role of ghrelin on diabetic neuropathic pain and its effect on the expression of purinergic receptor 2X-4/NOD-like receptor-3(P2X4/NLRP3).Methods:Forty male Sprague-Dawley rats were randomly allocated into 5 groups(n=8 each):normal+ghrelin group(group NG),normal+[D-Lys3]-GHRP-6 group(group ND),diabetes group(group D),diabetes+ghrelin group(group DG),and diabetes+ghrelin+[D-Lys3]-GHRP-6 group(group DGD).[D-Lys3]-GHRP-6 served as a ghrelin-specific inhibitor.Group NG received intraperitoneal injections of 200μg/kg ghrelin for 6 weeks,and group ND received intraperitoneal injections of 50 mg/kg[D-Lys3]-GHRP-6 for 6 weeks.Diabetes mellitus was induced by intraperitoneal injections of 60 mg/kg 1%streptozotocin.Three days after establishment of diabetes model,group DG was intraperitoneally given 200μg/kg ghrelin and group DGD given 200μg/kg ghrelin plus 50 mg/kg[D-Lys3]-GHRP-6 for 6 weeks.Mechanical withdrawal threshold(MWT)and motor nerve conduction velocity(MNCV)were measured at 2,4,and 6 weeks.After 6 weeks the rats were sacrificed.Rat spinal cords were examined for pathology change by Nissl staining and pathology change of the sural nerve by electron microscopy(EM).Expression of ghrelin,P2X4,NLRP3,and IL-1βin the spinal cord was detected by western blot.Results:Compared with group NG,group ND spinal Nissl staining and sural nerve EM showed normal spinal cell structure;there was no significant change in MWT and MNCV and no significant difference in the expression of P2X4,NLRP3,and IL-1β(P>0.05).Spinal Nissl staining and sural nerve EM of group D showed partial cellular damage,decreased MWT and MNCV,down-regulated expression of ghrelin,and up-regulated expression of P2X4,NLRP3,and IL-1β(P<0.05).Compared with group D,group DG showed normal spinal cell structure,increased MWT and MNCV,up-regulated expression of ghrelin,and down-regulated expression of P2X4,NLRP3,and IL-1β(P<0.05).Compared with group DG,group DGD showed severe cell structure damage,decreased MWT and MNCV,down-regulated expression of ghrelin,and up-regulated expression of P2X4,NLRP3,and IL-1β(P<0.05).Conclusion:Ghrelin may be involved in diabetic neuropathic pain through the P2X4/NLRP3 signaling pathway.
作者
刘康
周芳
赵博
黄亚医
王雅枫
LIU Kang;ZHOU Fang;ZHAO Bo;HUANG Ya-yi;WANG Ya-feng(Department of Anesthesiology,Renmin Hospital ofWuhan University,Wuhan 430060,China)
出处
《神经损伤与功能重建》
2019年第8期379-382,共4页
Neural Injury and Functional Reconstruction
基金
湖北省自然科学基金(No.2017CFB267)
中央高校基本科研业务费专项基金(No.2042018kf0161)
