摘要
目的探究当归芍药散调控核转录因子-κB抑制蛋白激酶(IKK)/核因子κB抑制蛋白(IκBα)/核因子-κB(NF-κB)通路对老年性聋小鼠耳蜗螺旋神经节细胞凋亡的影响。方法将48只C57BL/6J小鼠随机分为老年性聋模型组(Model组)、低剂量当归芍药散组(中药-L组,3.2 g/kg生药)、高剂量当归芍药散组(中药-H组,6.4 g/kg生药)和高剂量当归芍药散+IKK/IκBα/NF-κB信号通路激活剂白藜芦醇(RES)组(中药-H+RES组,6.4 g/kg生药+43.33 mg/kg RES),每组12只小鼠;取12只2月龄的C57BL/6J小鼠作为对照组(Control组)。采用听性脑干反应(ABR)检测各组小鼠听阈值;HE染色观察小鼠耳蜗螺旋神经节的病理变化,并对小鼠耳蜗中回与底回的螺旋神经节细胞的数量进行计数;TUNEL染色法检测小鼠耳蜗螺旋神经节细胞的凋亡。ELISA法检测小鼠耳蜗组织中丙二醛(MDA)与超氧化物歧化酶(SOD)水平。qRT-PCR法检测小鼠耳蜗组织中IL-1β、IL-6与TNF-αmRNA水平。Western blot法检测小鼠耳蜗组织IKK/IκBα/NF-κB通路与凋亡相关蛋白的表达。结果Model组小鼠较Control组小鼠8 kHz、16 kHz、24 kH、32 kHz听阈值、耳蜗螺旋神经节组织TUNEL阳性细胞数、耳蜗组织MDA及IL-1β、IL-6与TNF-αmRNA水平、p-IκBα、凋亡蛋白(Bax、cleaved-Caspase-3)表达水平、IKK、NF-κB p65磷酸化水平均显著升高(均P<0.05),耳蜗螺旋神经节细胞数量、SOD水平、IκBα蛋白表达水平显著降低(均P<0.05),耳蜗螺旋神经节细胞的形态异常、排列疏松紊乱,发生空泡化,出现病理学损伤。中药-L组、中药-H组小鼠较Model组相应指标变化程度较轻(均P<0.05)。RES减弱了当归芍药散对老年性聋小鼠耳蜗螺旋神经节细胞凋亡的抑制作用。结论当归芍药散可能通过下调IKK/IκBα/NF-κB通路抑制老年性聋小鼠耳蜗螺旋神经节细胞凋亡。
Objective To investigate the effect of Danggui Shaoyao powder on apoptosis of cochlear spiral ganglion cells of age-related hearing loss mice by regulating the inhibitor of kappa B kinase(IKK)/inhibitory subunit of nuclear factor-kappa B alpha(IκBα)/nuclear factor-kappa B(NF-κB)pathway.Methods A total of 48 C57BL/6J mice were randomly divided into age-related hearing loss model group(model group),low-dose Danggui Shaoyao powder group(traditional Chinese medicine-L group,3.2 g/kg crude drug),high-dose Danggui Shaoyao powder group(traditional Chinese medicine-H group,6.4 g/kg crude drug),and high-dose Danggui Shaoyao powder+IKK/IκBα/NF-κB signaling pathway activator resveratrol(RES)group(traditional Chinese medicine-H+RES group,6.4 g/kg crude drug+43.33 mg/kg RES),with 12 mice in each group.Twelve 2-month-old C57BL/6J mice were selected as the control group.Auditory brain stem response(ABR)was used to detect the auditory threshold of mice in each group.HE staining was used to observe the pathological changes of spiral ganglions of the mouse cochlea,and the number of spiral ganglion cells in the middle and bottom gyrus of the mouse cochlea was counted.The apoptosis of spiral ganglion cells in mouse cochlea was detected by TUNEL staining.The levels of malondialdehyde(MDA)and superoxide dismutase(SOD)in mouse cochlear tissue were detected by ELISA.The mRNA levels of IL-1β,IL-6 and TNF-αin mouse cochlear tissues were detected by qRT-PCR.Western blot was used to detect the expression of IKK/IκBα/NF-κB pathway and apoptosis related proteins in mouse cochlear tissue.Results Compared with the control group,the auditory thresholds at 8 kHz,16 kHz,24 kHz,and 32 kHz,the number of TUNEL positive cells in the cochlear spiral ganglion tissue,the level of MDA,and the mRNA level of IL-1β,IL-6 and TNF-αin the cochlear tissue,the expression of p-IκBα,apoptosis protein(Bax,and cleaved-Caspase-3),and the phosphorylation levels of IKK and NF-κB p65 in the model group were significantly increased(all P<0.05),the number o
作者
祝晓宇
邓华
孙川惠
曹庆新
Zhu Xiaoyu;Deng Hua;Sun Chuanhui(Department of Otorhinolaryngoloy,the First Affiliated Hospital of Guizhou University of Traditional Chinese Medicine,Guiyang 550001,China)
出处
《华中科技大学学报（医学版）》
CAS
CSCD
北大核心
2024年第4期466-472,共7页
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
基金
贵州省中医药管理局中医药、民族医药科学技术研究项目(No.QZYY-2023-047,No.QZYY-2023-026)
贵州省“十四五”中医药
民族医药重点学科建设规划项目[No.QZYYZDXK(PY)-2021-06]。
