鸣安方抑制水杨酸钠诱导大鼠耳鸣的中枢炎症机制  

Ming An Decoction Inhibits Tinnitus Related Central Inflammatory Mechanisms Induced by Salicylates in Rats

作　　者：徐智辉 郑伟[1] 陈斯 林佳川 张剑宁[1] 霍岩 XU Zhihui;ZHENG Wei;CHEN Si;LIN Jiachuan;ZHANG Jianning;HUO Yan(Department of Otorhinolaryngology,Yueyang Integrated Traditional Chinese and Western Medicine Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 200437,China)

机构地区：[1]上海中医药大学附属岳阳中西医结合医院耳鼻喉科,上海200437

出　　处：《中华耳科学杂志》2025年第1期89-94,共6页Chinese Journal of Otology

基　　金：国家自然科学基金青年项目(82405492);国家自然科学基金面上项目(82474263);科技部重点研发计划(2023YFC2508402)。

摘　　要：目的探究鸣安方干预耳鸣大鼠内侧膝状体(medial geniculate body,MGB)小胶质细胞激活诱导肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)表达的中枢炎症机制。方法将24只6周龄Wistar大鼠随机分为对照组、水杨酸注射耳鸣组、鸣安方组、TNF-α抑制剂组各6只,采用惊跳反射前脉冲抑制(pre-pulse inhibition,PPI)及惊跳反射间歇前抑制(gap pre-pulse inhibition of the acoustic startle,GPIAS)检测各组大鼠给药前后行为学改变;取各组大鼠脑组织,定位MGB脑区,采用免疫荧光染色并通过骨架分析法计算大鼠MGB小胶质细胞分枝数(endpoints/cell,EP/C)及分枝长度(process length/cell,PL/C),评估细胞形态;采用实时荧光定量聚合酶链式反应(real time quantitative polymerase chain reaction,RT-qPCR)及蛋白质印迹检测各组大鼠MGB脑区TNF-αmRNA转录及TNF-α蛋白相对表达。结果干预后,各组大鼠PPI抑制率比较,差异无统计学意义(P>0.05)。水杨酸注射耳鸣组GPIAS抑制率低于对照组,差异有统计学意义(P<0.05);鸣安方组、TNF-α抑制剂组GPIAS抑制率均高于水杨酸注射耳鸣组,差异均有统计学意义(P<0.05)。水杨酸注射耳鸣组MGB内小胶质细胞EP/C、PL/C均低于对照组,差异有统计学意义(P<0.05);鸣安方组、TNF-α抑制剂组MGB小胶质细胞EP/C、PL/C均高于水杨酸注射耳鸣组,差异均有统计学意义(P<0.05);鸣安方组小胶质细胞EP/C、PL/C高于TNF-α抑制剂组,差异均有统计学意义(P<0.05)。水杨酸注射耳鸣组大鼠MGB内TNF-αmRNA基因和蛋白相对表达均高于对照组,差异有统计学意义(P<0.05);鸣安方组、TNF-α抑制剂组TNF-αmRNA基因和蛋白相对表达均低于水杨酸注射耳鸣组,差异有统计学意义(P<0.05);鸣安方组TNF-αmRNA基因表达低于TNF-α抑制剂组,差异有统计学意义(P<0.05),蛋白相对表达与TNF-α抑制剂组比较,差异无统计学意义(P>0.05)。结论鸣安方具有显著改善大鼠耳鸣行为学表现的作用,�Objective To investigate central inflammatory mechanisms of effects of Ming An Decoction on tinnitus related activation-induced TNF-αexpression in microglia of the medical geniculate body(MGB)in rats.Methods Twenty-four Wistar rats were randomly assigned to a control,a sodium salicylate treatment(SS),a Ming An Decoction treatment(MAD)and a TNF-αInhibitor treatment(QNZ)group.Behavioral changes were assessed by pre-pulse inhibition(PPI)and gap pre-pulse inhibition of the acoustic startle(GPIAS)modeling.Expression of the microglia-specific marker Iba-1 in the MGB was assessed using immunofluorescence(IF).As part of cell morphology evaluation,skeletal analysis was utilized to calculate endpoints/cell and process length/cell of MGB microglia.TNF-αmRNA transcription and protein expression levels in the MGB region were measured using real-time reverse transcription polymerase chain reaction(RT-qPCR)and Western Blot(WB).Results GPIAS inhibition in the SS group was lower than in the control group(P<0.05),and lower than in the QNZ and MAD groups(P<0.05).MGB microglia endpoints and branch length in the SS group were also lower than in the control group(P<0.05),and lower than in the MAD and QNZ groups(P<0.05),while MGB microglia endpoints and branch length in the MAD group were higher than in the QNZ group(P<0.05).mRNA transcription and protein expression levels of TNF-αin the MGB in the SS group were higher than in the control group(P<0.05),and higher than in the MAD and QNZ groups(P<0.05),while the mRNA transcription level of TNF-αin the MAD group was lower than in the QNZ(P<0.05)with and no significant difference in expression level of TNF-αprotein between the two groups(P>0.05).Conclusion Ming An Decoction may improve tinnitus associated behavioral performance in rats by inhibiting the activation of microglia,thereby down-regulating the expression of TNF-α.

关 键 词：耳鸣 内侧膝状体 鸣安方 小胶质细胞 肿瘤坏死因子-α 

分 类 号：R285.5[医药卫生—中药学]