基于PI3K/AKT信号通路探究芍药甘草汤对神经根型颈椎病模型大鼠Bax和Bcl-2蛋白表达的影响

目的:探究芍药甘草汤对神经根型颈椎病(CSR)模型大鼠血清磷脂酰肌醇3-激酶(PI3K)/丝氧酸-苏氨酸蛋白激酶(AKT)的调控作用,分析芍药甘草汤对CSR大鼠Bax及Bcl-2的影响。方法:随机将90只SPF级Wistar大鼠分为空白组、模型组、对照组、芍药甘草汤治疗低、中、高剂量组六组,每组15只。除空白组外,其他组以颈部神经根腋下鱼线卡压法构建建立CSR模型大鼠,采用大鼠步态障碍行为学评分进行模型评价。造模后7 d进行干预治疗,空白组和模型组采用生理盐水灌胃,对照组采用3MA干预,芍药甘草汤低、中、高剂量治疗组分别按8、12、16 g/kg剂量进行灌胃,分别干预治疗7 d。末次给药24 h后处死大鼠,取脊髓及神经根组织,酶联免疫吸附法测定各组大鼠丝氨酸残基的底物Bax和Bcl-2的表达水平; HE染色检测各组大鼠脊髓组织病理表现; 蛋白免疫印迹法(Western blot)检测各组大鼠脊髓组织中PI3K/AKT通路蛋白的表达情况。结果:造模后14 d,除空白组外,各组大鼠步态积分增高,提示造模成功; 给药14 d后,模型组大鼠血清中Bax表达明显增高,Bcl-2表达下降,神经根组织中PI3K、p-AKT蛋白表达降低,差异有统计学意义(P<0.05)。与模型组比较,对照组和芍药甘草汤各治疗组CSR大鼠在治疗后神经根积分明显降低,而且组织中Bax表达降低,Bcl-2表达升高(P<0.05); 神经根组织中PI3K、p-AKT蛋白表达水平显著升高(P<0.05),且芍药甘草汤治疗组大鼠具有剂量依赖性(P<0.01)。结论:芍药甘草汤可有效调控CSR模型大鼠PI3K、p-AKT蛋白表达,其机制可能与芍药甘草汤能够激活CSR大鼠组织中Bax和Bcl-2等生物因子抗细胞凋亡相关。

Exploring the effect of Shaoyao Gancao decoction on the expression of Bax and Bcl-2 protein in a rat model of cervical spondylotic radiculopathy based on PI3K/AKT signaling pathway

Objective:To explore the effects of Shaoyao Gancao decoction on serum phosphoinositide 3-kinase(PI3K)/seroxy-threonine protein kinase(AKT)in cervical spondylotic radiculopathy(CSR)model rats,analyze the effect of Shaoyao Gancao decoction on Bax and Bcl-2 in CSR rats.Methods:Ninety SPF Wistar rats were randomly divided into a blank group,a model group,and a control group,The Shaoyao Gancao decoction treated low,medium,and high-dose groups of 6 groups,each with 15 rats.Except for the blank group,the other groups used the neck nerve root axillary fishing line compression method to establish CSR model rats,and the gait disorder behavior score of rats was used for model evaluation.Intervention treatment was performed 7 days after modeling.The blank group and the model group were given normal saline intragastric administration,and the control group was given 3MA intervention.The low,medium and high doses of Shaoyao Gancao decoction were treated at 8,12 and 16 g/kg,respectively.The rats were intragastrically administered at a dose of 16 g/kg,and the patients were treated with intervention for 7 days.Rats were sacrificed 24 hours after the last administration,spinal cord and nerve root tissues were taken,and enzyme-linked immunosorbent assay was used to determine the expression levels of Bax and Bcl-2,the substrates of serine residues in each group HE staining was used to detect the spinal cord of each group Histopathological findings.Western blot was used to detect the expression of PI3K/AKT pathway protein in the spinal cord tissues of rats in each group.Results:14 days after modeling,except for the blank group,the gait score of each group increased,indicating that the modeling was successful.14 days after the administration,the expression of Bax in the serum of the model group increased significantly,the expression of Bcl-2 decreased,and the nerve roots the expression of PI3K and p-AKT protein in the tissues decreased,and the difference was statistically significant(P<0.05).Compared with the model group,the CSR rats in the control group and each treatment group of Shaoyao Gancao decoction significantly reduced the nerve root score after treatment,and the expression of Bax in the tissue decreased,and the expression of Bcl-2 increased(P<0.05).PI3K in the nerve root tissue the expression level of p-AKT protein was significantly increased(P<0.05),and the rats in the Shaoyao Gancao decoction treatment group were dose-dependent(P<0.01).Conclusion:Shaoyao Gancao decoction can effectively regulate the expression of PI3K and p-AKT protein in CSR model rats.The mechanism may be related to that Shaoyao Gancao decoction can activate Bax and Bcl-2 in the tissues of CSR rats to resist apoptosis.