Suining Central Hospital, Suining, 629000, China.
Senior Department of Otolaryngology-Head & Neck Surgery, The Sixth Medical Center of PLA General Hospital, Beijing, 100000, China.
Mol Biotechnol. 2024 Feb;66(2):321-331. doi: 10.1007/s12033-023-00755-6. Epub 2023 May 5.
To observe the expression changes of P2 protein in cochlear spiral ganglion cells before and after noise injury, and to explore the relationship between the changes of purinergic receptors in spiral ganglion cells and noise-induced hearing loss, so that the signal transduction of purinergic receptors can be used to treat SNHL The target point provides a theoretical basis. The experimental animals were randomly divided into normal and experimental groups. The experimental group was given 120 dB white noise continuous exposure for 10 days and 3 h a day. The auditory brainstem response was measured before and after the noise exposure. After the noise exposure, the two groups of animals were collected. Do immunofluorescence staining, western blot, fluorescence real-time quantitative PCR to observe the expression of P2 protein. The average hearing threshold of the animals in the experimental group increased to 38.75 ± 6.44 dB SPL after 7 days of noise exposure, and the high-frequency hearing loss was lower and severe; the average hearing threshold increased to 54.38 ± 6.80 dB SPL after 10 days of noise exposure, and the hearing loss at 4 k Hz was relatively high. Light; Frozen sections of cochlear spiral ganglion cells and staining of isolated spiral ganglion cells found that P2X2, P2X3, P2X4, P2X7, P2Y2, and P2Y4 proteins were all expressed in cochlear spiral ganglion cells before noise exposure. Among them, P2X3 expression increased and P2X4, the down-regulation of P2Y2 expression was statistically significant (P < 0.05); Western blot and real-time quantitative PCR detection results showed that the expression of P2X3 was significantly increased after noise exposure than before noise exposure (P < 0.05), and P2X4 and P2Y2 were expressed after noise exposure The amount was significantly lower than before noise exposure (P < 0.05). (Figure. 4). After noise exposure, the expression of P2 protein is upregulated or downregulated. By affecting the Ca cycle, the transmission of sound signals to the auditory center is blocked, which provides a theoretical basis for the signal transduction of purinergic receptors to become a target for the treatment of SNHL.
为观察噪声损伤前后耳蜗螺旋神经节细胞 P2 蛋白的表达变化,探讨螺旋神经节细胞嘌呤能受体变化与噪声性听力损失的关系,以期使嘌呤能受体信号转导成为治疗 SNHL 的靶点提供理论依据。实验动物随机分为正常组和实验组,实验组给予 120dB 白噪声连续暴露 10 天,每天 3 小时。噪声暴露前后进行听性脑干反应测试,噪声暴露后两组动物取材,行免疫荧光染色、western blot、荧光实时定量 PCR 观察 P2 蛋白的表达。实验组动物噪声暴露 7 天后平均听阈升高至 38.75±6.44dB SPL,高频听力下降且较重;噪声暴露 10 天后平均听阈升高至 54.38±6.80dB SPL,4kHz 听力较高;耳蜗螺旋神经节细胞冰冻切片及分离的螺旋神经节细胞染色发现,噪声暴露前 P2X2、P2X3、P2X4、P2X7、P2Y2、P2Y4 蛋白均在耳蜗螺旋神经节细胞表达,其中 P2X3 表达增加,P2X4、P2Y2 表达下调,差异有统计学意义(P<0.05);Western blot 及实时定量 PCR 检测结果显示,噪声暴露后 P2X3 的表达量较噪声暴露前明显增加(P<0.05),而 P2X4、P2Y2 的表达量较噪声暴露前明显降低(P<0.05)。(图 4)。噪声暴露后 P2 蛋白表达上调或下调,通过影响 Ca 循环,阻断声信号向听觉中枢的传递,为嘌呤能受体信号转导成为 SNHL 治疗靶点提供理论依据。