链脲佐菌素诱导的糖尿病模型小鼠热痛反应潜伏时间与坐骨神经髓鞘相关蛋白表达的相关性
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摘要
目的观察链脲佐菌素(streptozotocin,STZ)诱导的糖尿病模型小鼠热痛反应潜伏时间与坐骨神经髓鞘相关蛋白表达的相关性。方法 16只6周龄雄性SPF级C57BL/6J小鼠随机分成两组,每组8只。STZ处理组小鼠采用一次性腹腔注射STZ 200 mg/kg,建立小鼠糖尿病模型;对照组注射同等体积缓冲液。采用YLS-6A智能热板仪进行热痛反应潜伏期测量,记录各组小鼠热痛反应潜伏期。采用实时定量PCR检测两组小鼠坐骨神经髓鞘相关蛋白MAG、P0 mRNA的表达;采用western blot检测髓鞘相关蛋白MAG、P0的蛋白表达水平。并分析小鼠热痛反应潜伏时间与髓鞘相关蛋白表达水平的相关性。结果与对照组比较,STZ处理组小鼠FBG升高(P<0.0001),成功建立糖尿病模型。与对照组相比,STZ处理组小鼠热痛反应潜伏期延长(P<0.05),STZ组小鼠热痛不敏感。与对照组比较,STZ处理组小鼠坐骨神经髓鞘相关蛋白MAG、P0 m RNA(P<0.0001)以及蛋白水平的表达下降。结论 STZ诱导的糖尿病模型小鼠热痛反应时间延长,与坐骨神经髓鞘相关蛋白表达水平下降相关。但具体机制仍待进一步探讨。
ObjectiveTo observe correlation between thermal response latency and sciatic nerve myelin associated protein in streptozotocin-induced mouse with diabetes.MethodsSixteen 6-week-old male SPF grade C57BL/6J mice were randomly divided into two groups with 8 mice each.STZ 200mg/kg in STZ group was injected intraperitoneally once to establish the mouse model with diabetes.The same volume of buffer was injected in CON group.Mice were placed on a hot plate apparatus to value the thermal response latency.Real-time RT-qPCR and western blot were used to detect the expression of MAG,P0 in the sciatic nerve of mice.The correlation between the thermal response latency and MAG、P0 was analyzed.Results Compared with CON group,the FBG level of STZ group was significantly increased(P<0.0001),which showed that the STZ-induced diabetes model was successfully established.Compared with CON group,the thermal response latency of STZ group was longer(P<0.05).The m RNA and protein expression levels of MAG、P0 were all significantly decreased in group A than those in group B(P<0.0001).ConclusionThe sciatic nerve myelin associated proteins are attenuated in the STZ-induced mouse model with diabetes.The results suggest that peripheral hyperglycemia may cause thermalgesia,which might relative to the decrease of myelin associated protein.But the specific mechanism remains to be explored.
ObjectiveTo observe correlation between thermal response latency and sciatic nerve myelin associated protein in streptozotocin-induced mouse with diabetes.MethodsSixteen 6-week-old male SPF grade C57BL/6J mice were randomly divided into two groups with 8 mice each.STZ 200mg/kg in STZ group was injected intraperitoneally once to establish the mouse model with diabetes.The same volume of buffer was injected in CON group.Mice were placed on a hot plate apparatus to value the thermal response latency.Real-time RT-qPCR and western blot were used to detect the expression of MAG,P0 in the sciatic nerve of mice.The correlation between the thermal response latency and MAG、P0 was analyzed.Results Compared with CON group,the FBG level of STZ group was significantly increased(P<0.0001),which showed that the STZ-induced diabetes model was successfully established.Compared with CON group,the thermal response latency of STZ group was longer(P<0.05).The m RNA and protein expression levels of MAG、P0 were all significantly decreased in group A than those in group B(P<0.0001).ConclusionThe sciatic nerve myelin associated proteins are attenuated in the STZ-induced mouse model with diabetes.The results suggest that peripheral hyperglycemia may cause thermalgesia,which might relative to the decrease of myelin associated protein.But the specific mechanism remains to be explored.
引文
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[10]Tang W,Chen X,Liu H,et al.Expression of Nrf2 promotes schwann cell-mediated sciatic nerve recovery in diabetic peripheral neuropathy[J].Cell Physiol Biochem,2018,46(5):1879-1894.
[11]Tan W,Rouen S,Barkus KM,et al.Nerve growth factor blocks the glucose-induced down-regulation of caveolin-1 expression in Schwann cells via p75 neurotrophin receptor signaling[J].J Biol Chem,2003,278(25):23151-23162.2018-11-01
[2]Nakatani M,Sasaki H,Kurisu S,et al.,Numbness and paresthesia in bilateral toes and soles,and disproportional sweating restricted to face and trunk are suitable symptoms useful for the diagnosis of diabetic symmetric polyneuropathy[J].J Diabetes Investig,2011,2(6):464-473.
[3]Candrilli SD,Davis KL,Kan HJ,et al,Prevalence and the associated burden of illness of symptoms of diabetic peripheral neuropathy and diabetic retinopathy[J].J Diabetes Complications,2007,21(5):306-314.
[4]Pop-Busui R,Lu J,Brooks MM,et al.Impact of glycemic control strategies on the progression of diabetic peripheral neuropathy in the Bypass Angioplasty Revascularization Investigation 2Diabetes(BARI 2D)Cohort[J].Diabetes Care,2013,36(10):3208-3215.
[5]Rojewska E,Zychowska M,Piotrowska A,et al.Involvement of macrophage inflammatory protein-1 family members in the development of diabetic neuropathy and their contribution to effectiveness of morphine[J].Front Immunol,2018,9:494.
[6]Xu W,Manichella D,Jiang H,et al.Absence of P0 leads to the dysregulation of myelin gene expression and myelin morphogenesis[J].J Neurosci Res,2000,60(6):714-724.
[7]Kawashima R,Kojima H,Nakamura K,et al.Alterations in mRNA expression of myelin proteins in the sciatic nerves and brains of streptozotocin-induced diabetic rats[J].Neurochem Res,2007,32(6):1002-1010.
[8]Schachner M,Bartsch U.Multiple functions of the myelin-associated glycoprotein MAG(siglec-4a)in formation and maintenance of myelin[J].Glia,2000,29(2):154-165.
[9]Shettar A,Muttagi G.Developmental regulation of insulin receptor gene in sciatic nerves and role of insulin on glycoprotein P0in the Schwann cells[J].Peptides,2012,36(1):46-53.
[10]Tang W,Chen X,Liu H,et al.Expression of Nrf2 promotes schwann cell-mediated sciatic nerve recovery in diabetic peripheral neuropathy[J].Cell Physiol Biochem,2018,46(5):1879-1894.
[11]Tan W,Rouen S,Barkus KM,et al.Nerve growth factor blocks the glucose-induced down-regulation of caveolin-1 expression in Schwann cells via p75 neurotrophin receptor signaling[J].J Biol Chem,2003,278(25):23151-23162.2018-11-01