温通活血乳膏对2型糖尿病周围神经病变模型大鼠背根神经节ERK1/2及p-ERK1/2的影响
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摘要
目的:观察温通活血乳膏对糖尿病周围神经病变(diabetic peripheral neuropathy,DPN)大鼠背根神经节(dorsal root ganglion,DRG)中ERK1/2蛋白表达水平及其磷酸化表达的影响。方法:使用75只高糖高脂饲料喂养后小剂量链脲佐菌素35 mg/kg(STZ)腹腔注射而诱发的2型糖尿病大鼠模型,成模的糖尿病大鼠继续高糖高脂饲料饲养8周,制成糖尿病周围神经病变大鼠模型。造模成功后按体质量随机分为模型组、温通活血乳膏高剂量+二甲双胍组、温通活血乳膏中剂量+二甲双胍组、温通活血乳膏低剂量+二甲双胍组、二甲双胍组、辣椒碱+二甲双胍组连续给药4周后检测各组大鼠的坐骨神经传导速度及潜伏期、热缩足潜伏期、冷缩足持续时间,采用Western Blot及免疫组化法检测大鼠DRG中ERK1/2及p-ERK1/2的蛋白表达水平。结果:温通高、中、低+二甲双胍组较模型组感觉神经传导速度加快、感觉神经潜伏期缩短,热缩足潜伏期、冷缩足持续时间延长(P<0.05),DRG中p-ERK1/2蛋白表达量降低(P<0.05)。结论:温通活血乳膏可通过抑制DRG中ERK1/2发生磷酸化减轻糖尿病周围神经病变大鼠神经病理性疼痛。
Objective: To observe the effect of Wentong Huoxue Cream for diabetic peripheral neuropathy(diabetic peripheral neuropathy, DPN) in the rat dorsal root ganglion(dorsal root ganglion, DRG) ERK1/2 protein expression levels and its phosphorylation in expression. Methods:After using 75 high-sugar high-fat feed urea with small dose of chain rhzomorph 35 mg/kg(STZ) intraperitoneal injection to induce rat model of type 2 diabetes, diabetic rats were given high-sugar high-fat feed for eight weeks to make diabetic peripheral neuropathy. After the success of modeling,the rats according to body weight were randomly divided into model group, Wentong Huoxue Ceam high dose+ metformin group, Wentong Huoxue Cream middle dose+ metformin group, Wentong Huoxue Cream low dose + metformin group, the metformin group, capsaicin + metformin group. After four consecutive weeks, the incubation period between groups' sciatic nerve conduction velocity and duration of incubation period, heat shrinkable feet, cold feet were detected. Western Blot and immunohistochemical method were used to detect the DRG ERK1/2 and p-ERK1/2 protein expression levels. Results:Wentong Huoxue Cream high, medium and low dose+ metformin groups' sensory nerve conduction speed,incubation period, heat shrinkable foot incubation period and cold enough duration were shortened compared with those of the model group(P<0.05). P-ERK1/2 protein expression in DRG decreased(P<0.05). Conclusion:Wentong Huoxue Cream can reduce the pathological pain of neuropathy in rats with diabetic peripheral neuropathy by inhibiting the phosphorylation of ERK1/2 in DRG.
Objective: To observe the effect of Wentong Huoxue Cream for diabetic peripheral neuropathy(diabetic peripheral neuropathy, DPN) in the rat dorsal root ganglion(dorsal root ganglion, DRG) ERK1/2 protein expression levels and its phosphorylation in expression. Methods:After using 75 high-sugar high-fat feed urea with small dose of chain rhzomorph 35 mg/kg(STZ) intraperitoneal injection to induce rat model of type 2 diabetes, diabetic rats were given high-sugar high-fat feed for eight weeks to make diabetic peripheral neuropathy. After the success of modeling,the rats according to body weight were randomly divided into model group, Wentong Huoxue Ceam high dose+ metformin group, Wentong Huoxue Cream middle dose+ metformin group, Wentong Huoxue Cream low dose + metformin group, the metformin group, capsaicin + metformin group. After four consecutive weeks, the incubation period between groups' sciatic nerve conduction velocity and duration of incubation period, heat shrinkable feet, cold feet were detected. Western Blot and immunohistochemical method were used to detect the DRG ERK1/2 and p-ERK1/2 protein expression levels. Results:Wentong Huoxue Cream high, medium and low dose+ metformin groups' sensory nerve conduction speed,incubation period, heat shrinkable foot incubation period and cold enough duration were shortened compared with those of the model group(P<0.05). P-ERK1/2 protein expression in DRG decreased(P<0.05). Conclusion:Wentong Huoxue Cream can reduce the pathological pain of neuropathy in rats with diabetic peripheral neuropathy by inhibiting the phosphorylation of ERK1/2 in DRG.
引文
[1] 王星, 李彩娜, 申竹芳. 黄芩素对糖尿病周围神经病变保护作用机制研究进展[J]. 中国临床药理学杂志, 2018(1):70-72.
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[3] 张凤, 洪铭范. 糖尿病周围神经病变的发病机制和诊断技术的研究进展[J]. 医学综述, 2017(23):4715-4720.
[4] 黎伟华, 吴璟, 杨洋,等. 氧化槐果碱通过调节ERK1/2-COX-2-PGE2通路改善小鼠炎性疼痛作用机制研究[J]. 畜牧与饲料科学, 2017, 38(11):15-19.
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[6] Tejeda HA, Wu J, Kornspun AR, et al. Pathway- and Cell-Specific Kappa-Opioid Receptor Modulation of Excitation-Inhibition Balance DifferentiallyGates D1 and D2 Accumbens Neuron Activity[J]. Neuron, 2017, 93(1):147-163.
[7] Choi DC, Lee JY, Lim EJ,et al.p38MAPK and ERK activation in microglia by acupuncture relieves neuropathic pain after spinal cord injury in rats [J].ExpNeurol,2012,236(2):268-282.
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[9] Popiolek-Barczy K,Makuch W,Rojewska E,et al.Inhibition of intracellular signaling pathways NF-κB and MEK1/2 attenuates neuropathic pain development and enhances morphine analgesia[J].Pharmacol Rep,2014,66(5):845-881.
[10] 陈茜,王卫群,李凯利.温通活血乳膏的抗炎消肿、镇痛及改善微循环作用实验研究[J].中国民康医学,2006,18(7):565-566.
[11] 朱建红,张新军,吕安冲.温通活血乳膏联合激肽原酶肠溶片治疗糖尿病痛性神经病变临床研究「J].河南中医,2017,37(4):663-668.
[12] 袁伯俊,廖明阳,李波.药物毒理学实验法与技术[M].北京:化学工业出版社生物医药出版分社,2007:216-217.
[13] 徐叔云,卞如濂,陈修.药理实验方法学[M].3版.北京:人民卫生出版社,2002:202-204.
[14] 冷向阳,王旭凯,李振华.丹归消肿膏治疗急性闭合性软组织损伤的实验研究[J].中国医药指南,2011,9(36):70-71.
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[17] Teixeiro E, Daniels MA. ERK and cell death: ERK location and T cell selection[J]. FEBS.J, 2010,277(1): 30-38.
[18] Zehorai E, Yao Z, Plotnikov A, et al. The subcellular localization of MEK and ERK--a novel nuclear translocation signal (NTS) paves a way to the nucleus[J]. Mol Cell Endocrinol, 2010, 314(2): 213-220.
[19] 杜全宇, 王振兴, 孙中莉,等. HMGB1对人肺成纤维细胞ERK1/2-NF-κB信号通路的影响及补阳还五汤含药血清的干预作用[J]. 中国实验方剂学杂志, 2017(13):153-157.
[20] 黄葱葱, 陈果, 吴艳,等. p-ERK1/2-AP-1通路在姜黄素抗大鼠糖尿病神经病理性痛中的作用[J]. 中国病理生理杂志, 2011, 27(6):1143-1148.
[21] 张鸿程, 陈彪, 谢庆平,等. 鞘内注射ERK1/2抑制剂对骨癌痛大鼠的镇痛作用研究[J]. 医学研究杂志, 2017, 46(5):119-124.
[22] 雷载权,张廷模.中华临床中药学[M].北京:人民卫生出版社,1998:1428,1600.
[23] 许源, 宿树兰, 王团结,等. 桂枝的化学成分与药理活性研究进展[J]. 中药材, 2013, 36(4):674-678.
[2] Pop-Busui R, Boulton AJ, Feldman EL, et al. Diabetic Neuropathy: A Position Statement by the American Diabetes Association[J]. Diabetes Care, 2017, 40(1):136-154.
[3] 张凤, 洪铭范. 糖尿病周围神经病变的发病机制和诊断技术的研究进展[J]. 医学综述, 2017(23):4715-4720.
[4] 黎伟华, 吴璟, 杨洋,等. 氧化槐果碱通过调节ERK1/2-COX-2-PGE2通路改善小鼠炎性疼痛作用机制研究[J]. 畜牧与饲料科学, 2017, 38(11):15-19.
[5] Van den Heuvel I,Reichl S,Segelcke D,et al.Selective prevention of mechanical hyperalgesia after incision by spinal ERK1/2 inhibition[J].Eur J Pain,2015,19(2):225-235.
[6] Tejeda HA, Wu J, Kornspun AR, et al. Pathway- and Cell-Specific Kappa-Opioid Receptor Modulation of Excitation-Inhibition Balance DifferentiallyGates D1 and D2 Accumbens Neuron Activity[J]. Neuron, 2017, 93(1):147-163.
[7] Choi DC, Lee JY, Lim EJ,et al.p38MAPK and ERK activation in microglia by acupuncture relieves neuropathic pain after spinal cord injury in rats [J].ExpNeurol,2012,236(2):268-282.
[8] Calvo M,Zhu N,Grist J,et al. Following nerve injury neuregulinl drives microglial proliferation and neuropathic pain via the MEK/ERK pathway[J].Glia, 2011,59(4):554-568.
[9] Popiolek-Barczy K,Makuch W,Rojewska E,et al.Inhibition of intracellular signaling pathways NF-κB and MEK1/2 attenuates neuropathic pain development and enhances morphine analgesia[J].Pharmacol Rep,2014,66(5):845-881.
[10] 陈茜,王卫群,李凯利.温通活血乳膏的抗炎消肿、镇痛及改善微循环作用实验研究[J].中国民康医学,2006,18(7):565-566.
[11] 朱建红,张新军,吕安冲.温通活血乳膏联合激肽原酶肠溶片治疗糖尿病痛性神经病变临床研究「J].河南中医,2017,37(4):663-668.
[12] 袁伯俊,廖明阳,李波.药物毒理学实验法与技术[M].北京:化学工业出版社生物医药出版分社,2007:216-217.
[13] 徐叔云,卞如濂,陈修.药理实验方法学[M].3版.北京:人民卫生出版社,2002:202-204.
[14] 冷向阳,王旭凯,李振华.丹归消肿膏治疗急性闭合性软组织损伤的实验研究[J].中国医药指南,2011,9(36):70-71.
[15] 马丽,陈茜.温通活血乳膏联合依帕司他对糖尿病痛性神经病变的影响[J].中药药理与临床,2014,30(4):122-125.
[16] BodartJF.Extracellular-regulated kinase-mitogen-activated protein kinase cascade: unsolved issues[J]. J Cell Biochem, 2010,10S(5): 850-857.
[17] Teixeiro E, Daniels MA. ERK and cell death: ERK location and T cell selection[J]. FEBS.J, 2010,277(1): 30-38.
[18] Zehorai E, Yao Z, Plotnikov A, et al. The subcellular localization of MEK and ERK--a novel nuclear translocation signal (NTS) paves a way to the nucleus[J]. Mol Cell Endocrinol, 2010, 314(2): 213-220.
[19] 杜全宇, 王振兴, 孙中莉,等. HMGB1对人肺成纤维细胞ERK1/2-NF-κB信号通路的影响及补阳还五汤含药血清的干预作用[J]. 中国实验方剂学杂志, 2017(13):153-157.
[20] 黄葱葱, 陈果, 吴艳,等. p-ERK1/2-AP-1通路在姜黄素抗大鼠糖尿病神经病理性痛中的作用[J]. 中国病理生理杂志, 2011, 27(6):1143-1148.
[21] 张鸿程, 陈彪, 谢庆平,等. 鞘内注射ERK1/2抑制剂对骨癌痛大鼠的镇痛作用研究[J]. 医学研究杂志, 2017, 46(5):119-124.
[22] 雷载权,张廷模.中华临床中药学[M].北京:人民卫生出版社,1998:1428,1600.
[23] 许源, 宿树兰, 王团结,等. 桂枝的化学成分与药理活性研究进展[J]. 中药材, 2013, 36(4):674-678.