基于PI3K/Akt1通路探讨补气养血通络组方改善硼替佐米所致大鼠周围神经毒性  

Effect of Buqi Yangxue Tongluo decoction on bortezomib-induced peripheral neurotoxicity in rats based on PI3K/Akt1 pathway

作　　者：王苗 马芹[1] 王志芳[1] 张萌[1] WANG Miao;MA Qin;WANG Zhifang;ZHANG Meng(Department of Radiochemotherapy,Cangzhou Hospital of Integrated Traditional Chinese and Western Medicine,Cangzhou 061000,China)

机构地区：[1]河北省沧州中西医结合医院放化疗科,河北沧州061000

出　　处：《生物技术》2024年第6期767-773,721,共8页Biotechnology

基　　金：河北省中医药管理局科研计划项目(2021341)。

摘　　要：[目的]基于PI3K/Akt1通路探讨补气养血通络组方对硼替佐米(Vel)所致大鼠周围神经毒性(BiPN)的作用,明确其作用机制。[方法]选取36只成年雄性SD大鼠,随机分为空白组,硼替佐米组(Vel组)、甲钴胺组、补气养血通络组低、中、高剂量组,每组6只。除空白组外,其余各组大鼠皮下注射硼替佐米(1 mg/kg)每周给药2次持续给药32 d建立BiPN大鼠模型。同时甲钴胺组给予甲钴胺与0.9%氯化钠注射液混悬液(0.05 mg/mL)灌胃;补气通络养血汤低、中、高剂量组给予1.0 g/mL、2.5 g/mL、4.0 g/mL的补气通络养血汤灌胃;空白组及Vel组给予0.9%氯化钠注射液灌胃。通过行为学检测大鼠冷刺激敏感度与机械性缩足阈值;通过HE染色分析大鼠背根神经节病理变化;使用酶联免疫吸附实验(ELISA)检测大鼠血清中SIRT3、IL-6、TNF-α水平以及实时荧光定量聚合酶链反应(RT-qPCR)检测大鼠脊髓中PI3K、Akt1、mTOR mRNA相对表达量。[结果]给药17 d,补气通络养血汤高剂量组冷刺激敏感度与机械性缩足阈值高于Vel组(17.83±2.01 vs 14.17±1.34;6.26±0.64 vs 5.38±0.28;P<0.05)。给药25 d,与Vel组相比,甲钴胺组和补气通络养血汤高剂量组冷刺激敏感度与机械性缩足阈均升高(P<0.05)。给药32 d,与Vel组相比,甲钴胺组、补气通络养血汤(中、高剂量)组冷刺激敏感度与机械性缩足阈均升高(P<0.05);与空白组相比,Vel组SIRT3水平降低,IL-6及TNF-α水平升高(116.73±5.39 vs 77.68±4.17;78.42±6.36 vs 104.27±7.28;40.03±8.36 vs 97.29±10.68;P<0.05)。与Vel组相比,甲钴胺组及补气通络养血汤(低、中、高剂量组)背根神经节损伤逐渐恢复、SIRT3水平均有所上升、IL-6及TNF-α水平均降低,但仅甲钴胺组、补气通络养血汤(中、高剂量组)与Vel组相比具有统计意义(P<0.05);与空白组相比,Vel组PI3K、AKT1、mTOR mRNA相对表达量均降低(2.89±0.84 vs 0.72±0.41;2.77±0.54 vs 0.62±0.61;2.93±0.68 vs 0.71±0.56;P<0.05)�[Objective]To study the effect of Buqi Yangxue Tongluo decoction on bortezomib(Vel)-induced peripheral neu-rotoxicity(BiPN)in rats based on PI3K/Akt1 pathway,and to clarify its mechanism.[Method]Thirty-six adult male SD rats were selected and randomly divided into blank group,bortezomib group(Vel group),methylcobalamin group,low-,medium-and high-dose Buqi Yangxue Tongluo decoction groups,six in each group.Model of BiPN was established by subcutaneous in-jection of bortezomib(1 mg/kg)twice a week for 32 d in all groups except blank group.After model establishment,methylco-balamin group was given methylcobalamin with 0.9%sodium chloride injection suspension(0.05 mg/mL)by gavage.Low-,medium-and high-dose groups were given 1.0 g/mL,2.5 g/mL and 4.0 g/mL Buqi Yangxue Tongluo decoction by gavage,whereas blank group and Vel group were given 0.9%sodium chloride injection by gavage.The sensitivity threshold to selective mechanical and cold stimulation was obtained using behavioural testing.SIRT3,IL-6,TNF-αin serum were measured using enzyme-linked immunosorbent assay(ELISA),and the relative mRNA expression of PI3K,Akt1,and mTOR in spinal cord was detected by real-time fluorescence quantitative polymerase chain reaction(RT-qPCR).[Result]After 17 days of adminis-tration,the sensitivity threshold to cold and selective mechanical stimulation of high-dose group were larger than those of Vel group(17.83±2.01 vs 14.17±1.34;6.26±0.64 vs 5.38±0.28;P<0.05).After 25 days of administration,an increase in the sensitivity threshold to cold and selective mechanical stimulation was observed in both mecobalamine group and high-dose group compared with Vel group(P<0.05).After 32 days of administration,the sensitivity threshold to cold and selective me-chanical stimulation were elevated in methylcobalamin group,medium-dose group and high-dose group compared with Vel group(P<0.05).Vel group had lower levels of SIRT3 and higher levels of IL-6 and TNF-αcompared to blank group(116.73±5.39 vs 77.68±4.17;78.42±6.36 vs 104.27±7.28;40.03�

关 键 词：补气养血通络汤 硼替佐米 周围神经毒性 PI3K/Akt1通路 

分 类 号：R745.7[医药卫生—神经病学与精神病学]