摘要
目的 探讨益肾舒筋汤对腰椎间盘突出症(LDH)大鼠的影响及作用机制。方法 采用自体髓核移植法构建LDH大鼠模型,将实验大鼠随机分为空白组,假手术组,模型组,益肾舒筋汤低、中、高剂量组。益肾舒筋汤组给予2、10、20g/ml的药物灌胃治疗21天。酶联免疫吸附法(ELISA)检测各组大鼠血清中肝肾功能指标谷丙转氨酶(ALT)、谷草转氨酶(AST)、肌酐(CRE)、尿素氮(BUN)、尿酸(UA)水平以及炎性因子白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)的含量;Von Frey纤维丝方法测定大鼠机械刺激缩足反射阈值(PMWT);热痛测试仪检测大鼠热刺激缩足反射潜伏期(PWTL);Western blot方法检测大鼠脊髓组织中微管相关蛋白轻链3B(LC3B)、自噬相关蛋白(Beclin-1)和泛素结合蛋白(p62)的蛋白表达。在高剂量益肾舒筋汤治疗的LDH模型大鼠中加入自噬抑制剂3-甲基腺嘌呤(3-MA)灌胃,检测3-MA对LDH大鼠的影响。结果 益肾舒筋汤对大鼠的肝肾功能没有明显影响。与假手术组比较,模型组大鼠的PWTL、PMWT降低,血清中炎性因子IL-1β、TNF-α、IL-6含量升高,脊髓组织中LC3B-Ⅱ/LC3B-Ⅰ、Beclin-1蛋白表达升高,p62的蛋白表达下降(P<0.05)。益肾舒筋汤干预后,与模型组比较,治疗组大鼠的PMWT、PWTL升高,炎性因子降低,LC3B-Ⅱ/LC3B-Ⅰ、Beclin-1蛋白表达进一步提升,p62蛋白表达降低,且变化在高剂量药物治疗组中更显著(P<0.05)。与舒筋汤组比较,舒筋汤+3-MA组小鼠自噬水平降低,PMWT、PWTL下降,血清中炎性因子水平提升(P<0.05)。结论 益肾舒筋汤可能通过激活自噬提高LDH大鼠疼痛阈值,降低炎症水平,对LDH大鼠起到治疗作用。
Objective To investigate the effect and mechanism of Yishenshujin Decoction on rats with lumbar disc herniation(LDH).Methods LDH rat model was established by autologous nucleus pulposus transplantation.The experimental rats were randomly divided into normal group,control group,model group,Yishenshujin decoction low-dose,medium-dose and high-dose groups.Yishenshujin Decoction group was given 2,10,20g/ml intragastric treatment for 21 days.Serum levels of AST,ALT,CRE,BUN,UA and inflammatory factors interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)were detected by Enzyme-Linked Immuno Sorbent Assay(ELISA).The paw mechanical withdrawal threshold(PMWT)was determined by Von Frey fiber.The paw withdrawal thermal latency(PWTL)was measured by heat pain tester.Protein expression of microtubule-associated protein 1 light chain 3B(LC3B),Beclin-1 and sequestosome 1(p62)in rat spinal tissue was measured by Western blot.The effect of autophagy inhibitor 3-Methyladenine(3-MA)on LDH rats treated with Yishenshujin Decoction was detected.Results Yishenshujin Decoction had no obvious effect on liver and kidney function in rats.Compared with control group,PMWT and PWTL were decreased,serum inflammatory factors IL-1β,TNF-αand IL-6 contents were increased,protein expression of LC3B-Ⅱ/LC3B-Ⅰand Beclin-1 in spinal cord tissue was increased,and protein expression of p62 was decreased in model group(P<0.05).After the intervention with Yishenshujin Decoction,PWTL and PMWT were increased,inflammatory factors were decreased,LC3B-Ⅱ/LC3B-Ⅰand Beclin-1 protein expression were further increased,and p62 protein expression was decreased in treatment groups compared with model group,and the changes were more significant in the high-dose drug treatment group(P<0.05).The administration of autophagy inhibitor 3-MA significantly inhibited the therapeutic effect of Yishenshujin Decoction on LDH rats.Conclusion Yishenshujin Decoction may improve the pain threshold and reduce the inflammation level in LDH rats by activating autophagy,and play a therapeutic role in LDH rats.
作者
王明森
王业杨
李焕彬
江发华
李伟鹏
秦育滨
黄伟斌
WANG Mingsen;WANG Yeyang;LI Huanbin;JIANG Fahua;LI Weipeng;QIN Yubin;HUANG Weibin(Puning Hospital of Traditional Chinese Medicine,Puning 515343,China)
出处
《中国煤炭工业医学杂志》
2024年第6期553-559,共7页
Chinese Journal of Coal Industry Medicine
基金
广东省中医药局科研项目(编号:20211465)
关键词
益肾舒筋汤
腰椎间盘突出症
炎症
自噬
3-MA
Yishenshujin decoction
Lumbar disc herniation
Inflammation
Autophagy
3-MA
