【摘要】 【目的】探讨补肾开窍方对6-羟基多巴(6-OHDA)诱导的帕金森病(PD)模型大鼠的神经保护作用及可能机制。【方法】将60只SD大鼠随机分为6组,即假手术组、模型组、石菖蒲挥发油组、龟板组、补肾开窍方组、IRE1抑制剂组,每组10只。采用大鼠左脑立体注射6-OHDA法建立PD大鼠模型。造模结束后,石菖蒲挥发油组大鼠给予30 mg/kg石菖蒲挥发油灌胃,龟板组大鼠给予3.77 g/kg龟板液灌胃,补肾开窍方组给予3.77 g/kg龟板液、30 mg/kg石菖蒲挥发油灌胃,IRE1抑制剂组大鼠给予15 mg/kg STF-083110腹腔注射,每日2次,给药30 d。高效液相色谱(HPLC)法测定纹状体内单胺类神经递质多巴胺(DA)的含量,流式细胞术测定中脑内酪氨酸羟化酶(TH)的表达率,蛋白免疫印迹(Western blot)法测定纹状体内葡萄糖调节蛋白78(GRP78)、C/EBP家族同源蛋白(CHOP)、磷酸化需肌醇酶1(p-IRE1)和X盒结合蛋白1(XBP1)的表达。【结果】与假手术组比较,模型组大鼠的自主运动功能、平衡能力与协调性及感觉运动整合功能显著降低(P<0.05),单胺类神经递质DA和TH表达显著降低(P<0.05),内质网应激(ERS)指标GRP78、CHOP表达及纹状体内p-IRE1、XBP1表达均升高(P<0.05)。补肾开窍方可使大鼠的行为学改善,增加脑内DA和TH含量,使ERS指标GRP78、CHOP表达降低,降低纹状体p-IRE1和XBP1水平[与IRE1抑制剂组比较无显著性差异(P>0.05)]。【结论】补肾开窍方可能通过抑制ERS的IRE1/XBP1通路对6-OHDA诱导的PD模型大鼠产生神经保护作用。更多还原

【Abstract】 Objective To explore the neuroprotective effects and mechanism of Bushen Kaiqiao Recipe(BKR)against Parkinson’s disease(PD)rats induced by 6-hydroxydopamine(6-OHDA). Methods Sixty SD rats were randomly divided into 6 groups,namely sham-operation group,model group,Acorus tatarinowii Schott volatile oil(AO)group,Tortoiseshell group,BKR group,and inositol requiring enzyme 1(IRE1)inhibitor group,10 rats in each group. PD rat model was established by left brain stereotactic injection of 6-OHDA. After modeling,AO group was given intragastric administration of 30 mg/kg of AO, Tortoiseshell group was given intragastric administration of 3.77 g/kg of Tortoiseshell decoction,BKR group was given intragastric administration of 30 mg/kg of AO combined with 3.77 g/kg of Tortoiseshell decoction,IRE1 inhibitor group was given intraperitoneal injection of 15 mg/kg of STF-083110,and sham-operation group and model group were given intragastric administration of 1 mL/100 g of distilled water,twice a day,the treatment covering 30 d. High performance liquid chromatography(HPLC)was used to detect the content of neurotransmitter of dopamine(DA)in corpus striatum,flow cytometry was used to measure the expression rate of tyrosine hydroxylase(TH),and Western blotting method was used to determine the expression levels of 78-kDa glucose-regulated protein(GRP78), C/EBP-homologous protein(CHOP),phosphorylated inositol requiring enzyme 1(p-IRE1)and X-box binding protein 1(XBP1). Results Compared with the sham-operation group, the autonomous movement, balance and coordination, and sensory motor integration function were decreased(P<0.05),DA content and TH expression rate were decreased(P<0.05), and the expression levels of endoplasmic reticulum stress(ERS)indicators of GRP78 and CHOP,and p-IRE1, XBP1 in corpus striatum were increased compared with those in the model group(P<0.05). After treatment, the behaviors of rats in BKR group were improved, DA content and TH expression rate were increased, the ERS indicators were lowered, the levels of p-IRE1 and XBP1 were decreased [being not significantly different from those in the IRE1 inhibitor group(P>0.05)]. Conclusion BKR has neuroprotective effects against 6-OHDA-induced parkinson’s disease in rats through inhibiting IRE1/XBP1 pathway.更多还原