Inhibition of the HMGB1/RAGE axis protects against cisplatin-induced ototoxicity via suppression of inflammation and oxidative stress.

As an anti-tumor drug widely used in the clinic, cisplatin is limited by its ototoxic side effects associated with various factors, including inflammatory responses. Receptor for Advanced Glycation Endproducts (RAGE) recognizes damage-associated molecular patterns (DAMPs) and promotes stress and inflammation. This study intended to determine the potential behavior of the HMGB1/RAGE axis after cisplatin injury and whether it has a protective effect after inhibiting this pathway. We used FPS-ZM1, a RAGE inhibitor, to modulate the axis of HMGB1/RAGE in neonatal mouse cochlear explants and C57BL/6 mice in vivo. Apoptosis was identified by Annexin V-FITC/PI assay, Cleaved Caspase-3, and TUNEL staining. Reactive oxygen species (ROS) level was assessed by MitoSOX Red and CellROX Green assay. The expression of proteins associated with the HMGB1/RAGE axis and apoptosis was observed by western blotting. The expression of inflammatory cytokines was evaluated by qPCR. The protective effect of HMGB1/RAGE knockdown was also assessed on cisplatin-induced ototoxicity. These results demonstrated that cisplatin could activate the HMGB1/RAGE pathway in cochlear hair cells and release inflammatory factors. Pretreatment with FPS-ZM1 alleviated cisplatin-induced ototoxicity in vivo and in vitro. Knocking down HMGB1 and RAGE achieved specific protective effects. Altogether, inhibiting HMGB1/RAGE axis can reverse the increase of ROS accumulation, the activation of apoptosis, and the production of inflammatory reactions after cisplatin injury. FPS-ZM1 could resist the ototoxicity of cisplatin by suppressing the HMGB1/RAGE signal pathway, and it may be considered the new otoprotective potential strategy for hearing loss.

Cinchonine and cinchonidine alleviate cisplatin-induced ototoxicity by regulating PI3K-AKT signaling.

AIM: Cinchonine (CN) and its isomer cinchonidine (CD), two of the common cinchona alkaloids, are wildly used as antimalarial drugs. However, the effects of CN and CD on the auditory system are unknown. METHODS: Molecular docking and molecular dynamics (MD) simulation were used for predicting effective drugs. The CCK-8 assay was conducted for assessing cell viability in House Ear Institute-Organ of Corti 1 (HEI-OC1) cells. MitoSox Red staining revealed reactive oxygen species (ROS) amounts. TMRM staining was used to assess the mitochondrial membrane potential (ΔΨm). Immunofluorescence staining of myosin 7a was used to examine hair cells (HCs) in cisplatin-treated neonatal mouse cochlear explants, while TUJ-1 immunostaining was used for the detection of spiral ganglion neurons (SGNs). Cleaved caspase-3 and TUNEL immunostaining were utilized for apoptosis assessment. Immunoblot was carried out to detect PI3K-AKT signaling effectors. RESULTS: Pretreatment with CN or CD significantly increased cell viability and reduced mitochondrial dysfunction and ROS accumulation in cisplatin-treated HEI-OC1 cells. Immunofluorescent staining of cochlear explants showed that CN and CD attenuated cisplatin-induced damage to SGNs and HCs. Immunoblot revealed that CN and CD downregulated the expression of cleaved caspase-3 and activated PI3K-AKT signaling in cisplatin-injured HEI-OC1 cells. CONCLUSION: CD and CN can reduce ototoxicity caused by cisplatin and might help treat cisplatin-associated hearing loss.

20(S)-Ginsenoside Rh1 inhibits cisplatin-induced hearing loss by inhibiting the MAPK signaling pathway and suppressing apoptosis in vitro.

As an anticancer drug, cisplatin is widely used, but its clinical application is restricted due to its severe side effects of ototoxicity. Therefore, this study was dedicated to assessing the benefit of ginsenoside extract, 20(S)-Ginsenoside Rh1 (Rh1), on cisplatin-induced ototoxicity. HEI-OC1 cells and neonatal cochlear explants were cultured. Cleaved caspase-3, TUNEL, and MitoSOX Red were observed in vitro by immunofluorescence staining. CCK8 and LDH cytotoxicity assays were detected to measure cell viability and cytotoxicity. Our results showed that Rh1 significantly increased cell viability, reduced cytotoxicity, and alleviated cisplatin-induced apoptosis. In addition, Rh1 pretreatment decreased the excessive accumulation of intracellular reactive oxygen species. Mechanistic studies indicated that Rh1 pretreatment reversed the increase of apoptotic protein expression, accumulation of mitochondrial ROS, and activation of the MAPK signaling pathway. These results suggested that Rh1 can act as an antioxidant and anti-apoptotic agent against cisplatin-induced hearing loss by suppressing the excessive accumulation of mitochondrial ROS, activation of MAPK signaling pathway and apoptosis.

5,7-Dihydroxy-4-methylcoumarin modulates the JNK/FoxO1 signaling pathway to attenuate cisplatin-induced ototoxicity by suppressing oxidative stress and apoptosis in vitro.

5,7-Dihydroxy-4-methylcoumarin (D4M) is attributed to free radical scavenging effects, with wide application for anti-oxidation. This work aimed to assess D4M's impact on cisplatin-induced ototoxicity. The cell viability was estimated with CCK-8 assay. Apoptosis was detected by the Annexin V-FITC and PI assay. The reactive oxygen species (ROS) level was determined by MitoSOX-Red and CellROX-Green probes. Mitochondrial membrane potential was analyzed with TMRM staining. Immunofluorescence was utilized for hair cells and spiral ganglion neuron detection. Apoptosis-associated proteins were assessed by cleaved caspase-3 and TUNEL staining. These results showed that D4M pretreatment protected hair cells from cisplatin-induced damage, increased cell viability, and decreased apoptosis in House Ear Institute-Organ of Corti1 (HEI-OC1) cells and neonatal mouse cochlear explants. D4M significantly inhibited cisplatin-induced mitochondrial apoptosis and reduced ROS accumulation. In addition, the protective effect of D4M on cisplatin-induced ototoxicity was also confirmed in cochlear hair cells and spiral ganglion neurons in neonatal mice. Mechanistic studies showed that D4M markedly downregulated p-JNK and elevated the expression ratio of p-FoxO1/FoxO1, thereby reducing cisplatin-induced caspase-dependent apoptosis. Meanwhile, D4M-related protection of HEI-OC1 cells was significantly blunted by JNK signaling induction with anisomycin. This study supports the possibility that D4M may be used as a new compound to prevent cisplatin-related hearing loss.

Vulnerable frequency as an independent prognostic factor for sudden sensorineural hearing loss.

Objectives: Sudden sensorineural hearing loss (SSNHL) is a common otology emergency in the practice. Its severe hearing impairment and prognosis impair the quality of life. Given that cochlear hair cell vulnerability is not consistent across frequencies, this study aims to investigate the impact of frequency-specific hearing loss on prognosis in SSNHL. Methods: The study included 255 patients with full-frequency SSNHL. The baseline, clinical, and hearing characteristics, as well as possible cardiovascular predictors in blood, were collected for analysis. Results: The 4,000 and 8,000 Hz hearing levels in the responder group were significantly lower than those in the non-responder group (p = 0.008, p < 0.001), while the average hearing was not (p = 0.081). Logistic regression showed that only vertigo (OR, 95% CI, 0.265, 0.102-0.684, p = 0.006) and 8,000 Hz hearing level (OR, 95% CI, 0.943, 0.916-0.971, p < 0.001) were strongly associated with treatment outcome. Conclusions: Compared with other frequencies, 8,000 Hz hearing level was closely related to prognosis in SSNHL. In an adjusted model, our study did not find an effect of mean hearing on prognosis in SSNHL. However, further multicenter prospective studies are needed for validation.

Automatic recognition of uterine contractions with electrohysterogram signals based on the zero-crossing rate.

Uterine contraction (UC) is an essential clinical indicator in the progress of labour and delivery. Electrohysterogram (EHG) signals recorded on the abdomen of pregnant women reflect the uterine electrical activity. This study proposes a novel algorithm for automatic recognition of UCs with EHG signals to improve the accuracy of detecting UCs. EHG signals by electrodes, the tension of the abdominal wall by tocodynamometry (TOCO) and maternal perception were recorded simultaneously in 54 pregnant women. The zero-crossing rate (ZCR) of the EHG signal and its power were calculated to modulate the raw EHG signal and highlight the EHG bursts. Then the envelope was extracted from the modulated EHG for UC recognition. Besides, UC was also detected by the conventional TOCO signal. Taking maternal perception as a reference, the UCs recognized by EHG and TOCO were evaluated with the sensitivity, positive predictive value (PPV), and UC parameters. The results show that the sensitivity and PPV are 87.8% and 93.18% for EHG, and 84.04% and 90.89% for TOCO. EHG detected a larger number of UCs than TOCO, which is closer to maternal perception. The duration and frequency of UC obtained from EHG and TOCO were not significantly different (p > 0.05). In conclusion, the proposed UC recognition algorithm has high accuracy and simple calculation which could be used for real-time analysis of EHG signals and long-term monitoring of UCs.

Development of Electrohysterogram Recording System for Monitoring Uterine Contraction.

Uterine contraction (UC) is an important clinical indictor for monitoring uterine activity. The purpose of this study is to develop a portable electrohysterogram (EHG) recording system (called PregCare) for monitoring UCs with EHG signals. The PregCare consisted of sensors, a signal acquisition device, and a computer with application software. Eight-channel EHG signals, the tocodynamometry (TOCO) signal, and maternal perception were recorded simultaneously by the signal acquisition device controlled by the computer via Bluetooth. PregCare was firstly evaluated by a signal simulator. Its relative error (RE) and coefficient of variation (CV) were calculated, and its agreement with the commercial instrument PowerLab was assessed by Bland-Altman plots. After that, PregCare was applied to 20 pregnant women in a hospital to record their EHG signals. These EHG signals were preprocessed and segmented into UCs and non-UCs. Then, the EHG features corresponding to UCs and non-UCs were extracted, respectively, including power spectral density (PSD), root mean square (RMS), peak frequency (PF), median frequency (MDF), and sample entropy (SamEn). One-way ANOVA was employed to assess the difference between UCs and non-UCs. The results show that RE and CV were less than 8% and 0.03%, respectively, which indicated the high accuracy and repeatability of PregCare. The small differences of mean and standard deviation indicated the high agreement between PregCare and PowerLab. Besides, the PSD of UCs was much larger than non-UCs between 0 and 0.7 Hz. RMS of UCs was significantly larger than non-UCs (p < 0.05). PF and SamEn of UCs were significantly smaller than non-UCs (p < 0.05). In conclusion, the developed EHG recording system was able to record EHG signals reliably. It has the advantages of portability, low power consumption, and wireless transmission, which can be used for long-term monitoring of UCs and prediction of the preterm delivery.

Estimation of 8-Electrode Configuration for Recognition of Uterine Contraction with Electrohysterogram.

As the representative of electrical activity from uterine muscle, electrohysterogram (EHG) is recorded non-invasively by multiple electrodes positioned on the abdominal surface. The purpose of our paper is to estimate different electrode configurations for recognizing uterine contractions (UCs) with EHG signals. 8-electrode configuration was taken as an example to show our novel method with convolutional neural network (CNN) classification and score. The open accessed Icelandic 16-electrode EHG database was adopted in our study. With 8-electrode configuration, EHG signals corresponding to UCs and non-UCs were segmented and saved as image patches. The CNN was established and trained by thousands of EHG segments. The performance of CNN was evaluated by the area under curve (AUC) and accuracy of recognizing UCs and non-UCs. Seven different 8-electrode configurations were scored and ranked. It was found the 8-electrode configuration with 4 on the uterine fundus, 2 on the body and 2 on the cervix achieved the AUC of 0.766 and the highest score of 2.197. Among the configurations we have tried, it is concluded that the 8 electrodes in 4-2-2 configuration placed along the uterus as an upside-down pear could provide the most important information for recognition of UC based on our experiments.