摘要
目的:观察加味补肝汤对糖尿病大鼠坐骨神经结构和功能的影响,验证其治疗糖尿病周围神经病变的疗效并分析可能的机制。方法:①实验于2004-11/2005-08在中南大学湘雅医院中西结合研究所实验室进行。选用8周龄的健康雄性Wistar大鼠30只。②30只雄性Wistar大鼠被随机分为3组:模型组、加味补肝汤组和正常对照组,每组10只。模型组和加味补肝汤两组大鼠禁食12h后,一次性腹腔注射1%链脲佐菌素(50mg/kg,pH4.2～4.5,0.1mol/L柠檬酸钠-柠檬酸缓冲液配制)诱导糖尿病大鼠模型,72h后尾静脉采血测定血糖,凡血糖>16.7mmol/L者纳入实验;正常对照组大鼠则禁食12h后一次性腹腔注射相应容积即0.1mol/L的柠檬酸缓冲液。造模后1周加味补肝汤组按13.6g/(kg·d)剂量灌胃,加味补肝汤(由枸杞15g,木瓜15g,当归10g,川芎10g,熟地12g,白芍15g,桑寄生15g,麦冬15g,天花粉15g等组成,医院制剂,药物来源于中南大学湘雅医院药剂科)。模型组和正常组灌服等容积的蒸馏水(1mL/100g),1次/d,连续8周。③予以加味补肝汤治疗8周后,在光镜、电镜下观察坐骨神经形态改变;化学比色法分析血清丙二醛水平;采用MS302多媒体生物信号系统分别记录用药4和8周末大鼠坐骨神经传导速度。④计量资料差异比较采用t检验。结果:正常对照组无动物死亡,模型组、加味补肝汤组在用药4周末各死亡1只,8周末死亡2和3只。①坐骨神经形态结构变化:正常对照组大鼠坐骨神经有髓神经纤维形态正常;模型组出现轴突萎缩,髓鞘的变性;加味补肝汤组病变程度轻于模型组。②血清丙二醛水平:模型组和加味补肝汤组鼠明显高于正常对照组(t=3.594,8.513,P<0.01),模型组明显高于加味补肝汤组(t=8.513,3.607,P<0.01)。③坐骨神经传导速度:模型组和加味补肝汤组大鼠在用药4和8周末均明显慢于正常对照组(t=3.839～8.438,P<0.01),加味补肝汤组明显快于模型组(P<0.05)。④血糖:模型组与加味补肝汤组用药前和用药4和8周末差异不明显(P>0.05)。结论:加味补肝汤对糖尿病大鼠周围神经病变具有一定防治作用,但目前剂量末影响其血糖,说明其改善神经结构和功能的疗效不是直接通过降低血糖而实现,有可能是加味补肝汤通过有效清除自由基,减轻脂质过氧化而实现的。
AIM： To investigate the effects of jiawei bugan decoction （JWBGD） on structure and function of sciatic nerve in diabetic rats, and to manifest its therapeutic effects and analyze its possible mechanism on diabetic peripheral neuropathy.
METHODS： ① This experiment was completed in the Laboratory of Institute of Conthined Traditional Chinese and West Medicine, Xiangya Hospital, Central South University from November 2004 to August 2005. Totally 30 healthy male Wistar rats of eight-week old were selected. ② All male Wistar animals were randomly divided into 3 groups： model group, JWBGD group and normal control group with 10 in each group. After 12- hour fasting, rats in both model and JWBGD group were intraperitoneally injected with 1% streptozotocin （STZ） （50 mg/kg, pH4.2-4.5, 0.1 mol/L citrate buffer） to induce models. Vein blood was collected from tail to measure blood glucose after 72 hours. It blood glucose was greater than 16.7 mmol/L, rats were accepted in the study. Rats in normal control group were intraperitoneally injected with 0.1 mol/L citrate buffer after 12- hour fasting. One week after modeling, rats in JWBGD group were perfused with 13.6 g/（kg·d）.JWBGD. JWBGD was composed of gouqi 15 g, mugua 15 g, danggui 10 g, chuanxiong 10 g, shudi 12 g, ba/shao 15 g, sangjisheng 15 g, maidong 15 g, tianhuafen 15 g, etc. The decoction was provided by the hospital, and the herbs were provided by Pharmacy of Xiangya Hospital, Central South University. Isometric distilled water （1 mL/100 g） was perfused into rats in model group and normal control group, once a day. The perfusion had been executed continuously for 8 weeks. ③ After 8 weeks, various parameters were measured as follows： Morphometric evaluation of sciatic nerve was observed under light microscope and transmission electron microscope. MDA content in blood serum was analyzed by chromatometry, and the speed of conduction in nerve was recorded in 4^th and 8^th week using the MS302 style of media biosignal system.④Measurement data were compared with t test.
RESULTS： No animal died in the normal control group. One died in model group and JWBGD group at the end of the 4^th week respectively, and two and three died at the end of the 8^th weeks. ① Morphological changes of sciatic nerve： Rats in the normal control group had normal myelinated nerve fiber; but the atrophy of axis and the degeneration of myelin were observed in model group. Degree of lesion was lighter in JWBGD group than that in model group. ② MDA level： MDA level in model group and JWBGD group was higher than that in normal control group （t=3.594, 8.513, P 〈 0.01）, and that in model was higher than that in JWBGD group （t=8.513, 3.607, P 〈 0.01）.③ Conduction velocity of sciatic nerve： The velocity in model group and JWBGD group was slower than that in normal control group at the end Of the 4^th and 8^th weeks （t=3.839-8.438, P 〈 0.01）,and that in JWBGD group was faster than that in model group （P〈 0.05）. ④ Blood glucose： There was no significant difference between JWBGD group and model group before medication and at the end of the 4^th and 8^th weeks （P 〉 0.05）. CONCLUSION： JWBGD has prevention and therapeutic effects on peripheral neuropathy lesion of diabetic rats. However, dosage does not affect its blood glucose, this suggests that the effect on amelioration of neuromechanism and neural function is not realized by decreasing blood glucose; but by removing free radical, and relieving lipid peroxidation.
出处
《中国临床康复》
CSCD
北大核心
2006年第11期87-89,i0002,共4页
Chinese Journal of Clinical Rehabilitation
基金
湖南省卫生厅中医药重大研究项目(202002-3)~~
