摘要
目的:基于线粒体运输探讨补阳还五汤对糖尿病周围神经病变(DPN)的防治作用机制。方法:SD大鼠用高糖高脂饲料联合腹腔注射链脲佐菌素(STZ)诱导糖尿病。45只糖尿病大鼠随机分为DPN组、α-硫辛酸组、补阳还五汤组,每组15只,另设15只标准喂养大鼠为正常组。α-硫辛酸组、补阳还五汤组分别给与α-硫辛酸60 mg·kg^(-1)·d^(-1)、补阳还五汤15 g·kg^(-1)·d^(-1)灌胃治疗12周。给药结束后检测机械性痛阈(PWT)和运动神经传导速度(MNCV),取双侧坐骨神经及双侧L4-5的背根神经节。用50 mmol·L^(-1)葡萄糖和250μmol·L^(-1)棕榈酸钠干预NSC34细胞建立细胞损伤模型,随机分为空白组(10%空白血清)、DPN组(10%空白血清)、α-硫辛酸组(10%α-硫辛酸含药血清)、补阳还五汤组(10%补阳还五汤含药血清)、补阳还五汤加Compound C(CC)组(10%补阳还五汤含药血清+10μmol·L^(-1)CC),分别干预24 h。免疫荧光法、免疫组化法和蛋白免疫印迹法(Western blot)分别检测各实验组磷酸化单磷酸腺苷活化蛋白激酶(p-AMPK)、磷酸化环磷腺苷效应元件结合蛋白(p-CREB)及线粒体运输蛋白驱动家族蛋白成员5A(KIF5A)、细胞质动力蛋白1中间链2(DYNC1I2)的表达。结果:动物实验中,与正常组比较,DPN组大鼠空腹血糖显著升高(P<0.01),MNCV、PWT均显著降低(P<0.01);KIF5A、p-AMPK/AMPK、p-CREB/CREB显著降低(P<0.01),DYNC1I2表达显著升高(P<0.01)。与DPN组比较,α-硫辛酸组及补阳还五汤组大鼠MNCV、PWT显著升高(P<0.01);KIF5A、p-AMPK/AMPK、p-CREB/CREB明显升高(P<0.05,P<0.01);DYNC1I2明显降低(P<0.05,P<0.01)。与α-硫辛酸组比较,补阳还五汤组MNCV、KIF5A表达明显升高(P<0.05),DYNC1I2表达显著降低(P<0.01)。细胞实验中,与空白组比较,DPN组KIF5A、p-AMPK/AMPK、p-CREB/CREB显著降低(P<0.01),DYNC1I2表达显著升高(P<0.01)。与DPN组比较,α-硫辛酸组及补阳还五汤组KIF5A、p-AMPK/AMPK、p-CREB/CREB明显升高(P<0.05,P<0.0
Objective:To investigate the mechanism of Buyang Huanwutang in treating diabetic peripheral neuropathy(DPN)via mitochondrial transport.Method:Diabetes in SD rats was induced by a highcarbohydrate/high-fat diet and intraperitoneal injection of streptozotocin(STZ).The 45 diabetic rats were randomly assigned into a DPN group,an alpha-lipoic acid(60 mg·kg^(-1)·d^(-1))group,and a Buyang Huanwutang(15 g·kg^(-1)·d^(-1))group,with 15 rats in each group.Fifteen normal SD rats were fed with the standard diet and set as the control group.The rats were administrated with corresponding drugs by gavage for 12 weeks.The paw withdraw threshold(PWT)and motor nerve conduction velocity(MNCV)were measured at the end of medication,and the sciatic nerve and the bilateral dorsal root ganglia of L4-5 were collected.The injury model of NSC34 cells was established by treating with 50 mmol·L^(-1)glucose and 250μmol·L^(-1)sodium palmitate.The NSC34 cells were then randomly assigned into a blank(10%blank serum)group,a DPN(10%blank serum)group,an apha-lipoic acid(10%apha-lipoic acid-containing serum)group,a Buyang Huanwutang(10%Buyang Huanwutang-containing serum)group,and a Buyang Huanwutang+Compound C(CC)(10%Buyang Huanwutang-containing serum+10μmol·L^(-1)CC)group.The cell intervention lasted for 24 h.The immunofluorescence method,immunohistochemistry,and Western blot were employed to determine the expression levels of phosphorylated adenosine monophosphate-activated protein kinase(p-AMPK),phosphorylated cAMP-response element binding protein(p-CREB),kinesin family member 5A(KIF5A),and dynein cytoplasmic 1 intermediate chain 2(DYNC1I2).Result:Compared with the control group,the DPN group of rats showed increased fasting blood glucose(P<0.01),decreased MNCV and PWT(P<0.01),downregulated expression of KIF5A,p-AMPK/AMPK,and p-CREB/CREB(P<0.01),and up-regulated expression of DYNC1I2(P<0.01).Compared with the DPN group,drug intervention groups showed increased MNCV and PWT(P<0.01),up-regulated expression of KIF5A,p-AMPK/AMPK,and p-CREB/CR
作者
田佳鑫
安静文
张天雅
张志红
张旺
宋林春
段天梦
贲莹
TIAN Jiaxin;AN Jingwen;ZHANG Tianya;ZHANG Zhihong;ZHANG Wang;SONG Linchun;DUAN Tianmeng;BEN Ying(Hebei Key Laboratory of Integrative Medicine on Liver-Kidney Patterns,College of Integrated Traditional Chinese and Western Medicine,Hebei University of Chinese Medicine,Shijiazhuang 050091,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2023年第13期27-36,共10页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金青年基金项目(81803922)
河北省自然科学基金中医药联合基金培育项目(H2022423298)。
