糖尿病大鼠四肢痛阈及周围神经超微结构的变化
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摘要
目的观察链脲佐菌素(STZ)诱导Ⅰ型糖尿病大鼠模型四肢痛阈的变化及背根神经节细胞和腓肠神经超微结构变化。方法 6周龄清洁级雄性SD大鼠50只,随机分为对照组10只,模型组40只。模型组采用一次性腹腔注射60 mg/kg体质量的链脲佐菌素(STZ)诱导Ⅰ型糖尿病大鼠模型,对照组予以等量生理盐水,监测两组大鼠体质量及血糖变化,并观察大鼠痛阈改变;模型组分别于造模成功后2,4,6周随机选取8只大鼠取背根神经节和腓肠神经,采用透射电镜观察超微结构变化。结果大鼠糖尿病造模成功率为82.5%,随周龄增加,糖尿病组大鼠体型明显消瘦,体质量与对照组相比差异具有统计学意义(P <0.05);造模成功后第2周糖尿病组的机械性痛阈即低于正常组,且随时间的延长,痛阈逐渐减低,两组间差异具有统计学意义(P <0.05)。随病程增加背根神经节细胞核膜皱缩逐渐加重,内质网逐渐出现水肿,空泡化;腓肠神经脱髓鞘病变也逐渐加重,与机械痛阈的降低存在一致性。结论单次腹腔注射STZ(60 mg/kg)后可诱导出稳定的Ⅰ型糖尿病大鼠模型,且随病程增加,大鼠四肢痛阈降低逐渐加重,背根神经节细胞和腓肠神经超微结构病变也逐渐加剧。
Objective To observe the pain threshold and the ultrastructure of dorsal root ganglion cell and sural nerve in streptozotocin(STZ)-induced diabetic rats. Methods A total of 50 six-week-old male SD rats were randomly divided into control group(n = 10)and model group(n = 40). The mice in model group were intraperitoneally injected with STZ(Sigma,60 mg/kg body weight) to establish the diabetic model,while the mice in control group were given the same amount of saline. The changes of body weight,blood sugar and the mechanical withdrawal threshold(MWT) were monitored. Eight rats were randomly selected in model group after the successful modeling to observe the ultrastructural changes of dorsal root ganglion and sural nerve at weeks 2,4 and 6,respectively. Results All of the rats survived,and 82.5% rats had blood glucose more than 16.7 mmol/L. The body weight was significantly reduced in model group compared with control group at the same time point(P < 0.05). MWT was lower in model group than in control group at the second week after the successful modeling(P < 0.05),and it gradually decreased with the prolongation of time. With the development of diabetes,the shrinkage of nuclear membrane was aggravated gradually in the dorsal root ganglion,and the endoplasmic reticulum became edema and vacuolation gradually. In the sural nerve,the demyelinating lesions were also gradually aggravating,and the MWT was also decreased. Conclusion Stable type Ⅰ diabetic rat model can be induced by single intraperitoneal injection of STZ(60 mg/kg). With the progression of the disease,the pain threshold and the ultrastructural changes of dorsal root ganglion cells and sural nerve are aggravated gradually in rats.
Objective To observe the pain threshold and the ultrastructure of dorsal root ganglion cell and sural nerve in streptozotocin(STZ)-induced diabetic rats. Methods A total of 50 six-week-old male SD rats were randomly divided into control group(n = 10)and model group(n = 40). The mice in model group were intraperitoneally injected with STZ(Sigma,60 mg/kg body weight) to establish the diabetic model,while the mice in control group were given the same amount of saline. The changes of body weight,blood sugar and the mechanical withdrawal threshold(MWT) were monitored. Eight rats were randomly selected in model group after the successful modeling to observe the ultrastructural changes of dorsal root ganglion and sural nerve at weeks 2,4 and 6,respectively. Results All of the rats survived,and 82.5% rats had blood glucose more than 16.7 mmol/L. The body weight was significantly reduced in model group compared with control group at the same time point(P < 0.05). MWT was lower in model group than in control group at the second week after the successful modeling(P < 0.05),and it gradually decreased with the prolongation of time. With the development of diabetes,the shrinkage of nuclear membrane was aggravated gradually in the dorsal root ganglion,and the endoplasmic reticulum became edema and vacuolation gradually. In the sural nerve,the demyelinating lesions were also gradually aggravating,and the MWT was also decreased. Conclusion Stable type Ⅰ diabetic rat model can be induced by single intraperitoneal injection of STZ(60 mg/kg). With the progression of the disease,the pain threshold and the ultrastructural changes of dorsal root ganglion cells and sural nerve are aggravated gradually in rats.
引文
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[16]徐利娟,冯程程,岳薇薇,等.温通活血乳膏对糖尿病周围神经病变大鼠背根神经节中p-AKT蛋白表达的影响[J].中医杂志,2018,59(2):142-146.
[17]Al-Rejaie SS,Abuohashish HM,Ahmed MM,et al.Telmisartan inhibits hyperalgesia and inflammatory progression in a diabetic neuropathic pain model of Wistar rats[J].Neurosciences(Riyadh),2015,20(2):115-123.
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[19]Farshid AA,Tamaddonfard E.Histopathological and behavioral evaluations of the effects of crocin,safranal and insulin on diabetic peripheral neuropathy in rats[J].Avicenna J Phytomed,2015,5(5):469-478.
[20]Fatani AJ,Al-Rejaie SS,Abuohashish HM,et al.Neuroprotective effects of gymnema sylvestre on streptozotocin-induced diabetic neuropathy in rats[J].Exp Ther Med,2015,9(5):1670-1678.
[21]Hur J,Sullivan KA,Callaghan BC,et al.Identification of factors associated with sural nerve regeneration and degeneration in diabetic neuropathy[J].Diabetes Care,2013,36(12):4043-4049.
[22]吕翠岩,张胜容,徐暾海,等.糖痹康对糖尿病周围神经病变大鼠脂代谢及坐骨神经超微结构的影响[J].中华中医药杂志,2016,31(12):5000-5003.
[2]Jargen P,Dietrich A,Herling AW,et al.The role of insulin resistance in experimental diabetic retinopathy-Genetic and molecular aspects[J].PLo S One,2017,12(6):e178658.
[3]El-Sayed SS,Zakaria MN,Abdel-Ghany RH,et al.Cystathionine-gamma lyase-derived hydrogen sulfide mediates the cardiovascular protective effects of moxonidine in diabetic rats[J].Eur J Pharmacol,2016,783:73-84.
[4]王媛媛,贾立群,柯丹丹,等.奥沙利铂周围神经毒性模型大鼠行为学检测方法的比较[J].中国实验动物学报,2015,23(1):13-17.
[5]陈锐,王华,江伟.大鼠经寰枢椎蛛网膜下腔置管法与传统置管法的比较[J].中国比较医学杂志,2013,23(7):32-35.
[6]Kambiz S,van Neck JW,Cosgun SG,et al.An early diagnostic tool for diabetic peripheral neuropathy in rats[J].PLo S One,2015,10(5):e126892.
[7]Bouhassira D,Lanteri-Minet M,Attal N,et al.Prevalence of chronic pain with neuropathic characteristics in the general population[J].Pain,2008,136(3):380-387.
[8]李丹,陈龙菊.糖尿病足神经病变动物模型研究进展[J].中国实验动物学报,2018,26(1):128-132.
[9]O’Brien PD,Sakowski SA,Feldman EL.Mouse models of diabetic neuropathy[J].ILAR J,2014,54(3):259-272.
[10]Zhu H,Wang Y,Liu Z,et al.Antidiabetic and antioxidant effects of catalpol extracted from Rehmannia glutinosa(Di Huang)on rat diabetes induced by streptozotocin and high-fat,high-sugar feed[J].Chin Med,2016,11:25.
[11]Kim BJ,Nam JH,Kim KH,et al.Characteristics of gintoninmediated membrane depolarization of pacemaker activity in cultured interstitial cells of Cajal[J].Cell Physiol Biochem,2014,34(3):873-890.
[12]Sun W,Miao B,Wang XC,et al.Gastrodin inhibits allodynia and hyperalgesia in painful diabetic neuropathy rats by decreasing excitability of nociceptive primary sensory neurons[J].PLo SOne,2012,7(6):e39647.
[13]Chen SR,Pan HL.Hypersensitivity of spinothalamic tract neurons associated with diabetic neuropathic pain in rats[J].J Neurophysiol,2002,87(6):2726-2733.
[14]Menichella DM,Abdelhak B,Ren D,et al.CXCR4 chemokine receptor signaling mediates pain in diabetic neuropathy[J].Mol Pain,2014,10:42.
[15]刘伟,梁晓春,孙青,等.筋脉通对糖尿病大鼠背根神经节氧化应激及细胞凋亡的影响[J].中国医学科学院学报,2013,35(6):649-654.
[16]徐利娟,冯程程,岳薇薇,等.温通活血乳膏对糖尿病周围神经病变大鼠背根神经节中p-AKT蛋白表达的影响[J].中医杂志,2018,59(2):142-146.
[17]Al-Rejaie SS,Abuohashish HM,Ahmed MM,et al.Telmisartan inhibits hyperalgesia and inflammatory progression in a diabetic neuropathic pain model of Wistar rats[J].Neurosciences(Riyadh),2015,20(2):115-123.
[18]Jahanabadi S,Hadian MR,Shamsaee J,et al.The effect of spinally administered WIN 55,212-2,a cannabinoid agonist,on thermal pain sensitivity in diabetic rats[J].Iran J Basic Med Sci,2016,19(4):394-401.
[19]Farshid AA,Tamaddonfard E.Histopathological and behavioral evaluations of the effects of crocin,safranal and insulin on diabetic peripheral neuropathy in rats[J].Avicenna J Phytomed,2015,5(5):469-478.
[20]Fatani AJ,Al-Rejaie SS,Abuohashish HM,et al.Neuroprotective effects of gymnema sylvestre on streptozotocin-induced diabetic neuropathy in rats[J].Exp Ther Med,2015,9(5):1670-1678.
[21]Hur J,Sullivan KA,Callaghan BC,et al.Identification of factors associated with sural nerve regeneration and degeneration in diabetic neuropathy[J].Diabetes Care,2013,36(12):4043-4049.
[22]吕翠岩,张胜容,徐暾海,等.糖痹康对糖尿病周围神经病变大鼠脂代谢及坐骨神经超微结构的影响[J].中华中医药杂志,2016,31(12):5000-5003.