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This study aimed to evaluate the therapeutic potential of inhibiting protein arginine methyltransferase 5(PRMT5)in cisplatin-induced hearing loss.The effects of PRMT5 inhibition on cisplatin-induced auditory injury were determined using immunohistochemistry,apoptosis assays,and auditory brainstem response.The mechanism of PRMT5 inhibition on hair cell survival was assessed using RNA-seq and Cleavage Under Targets and Tagment-quantitative polymerase chain reaction(CUT&Tag-qPCR)analyses in the HEI-OC1 cell line.Pharmacological inhibition of PRMT5 significantly alleviated cisplatin-induced damage to hair cells and spiral ganglion neurons in the cochlea and decreased apoptosis by protecting mitochondrial function and preventing the accumulation of reactive oxygen species.CUT&Tag-qPCR analysis demonstrated that inhibition of PRMT5 in HEI-OC1 cells reduced the accumulation of H4R3me2s/H3R8me2s marks at the promoter region of the Pik3ca gene,thus activating the expression of Pik3ca.These findings suggest that PRMT5 inhibitors have strong potential as agents against cisplatin-induced ototoxicity and can lay the foundation for further research on treatment strategies of hearing loss.
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Cisplatin-related ototoxicity is a critical side effect of chemotherapy and can lead to irreversible hearing loss. This study aimed to assess the potential effect of the DNA methyltransferase (DNMT) inhibitor RG108 on cisplatin-induced ototoxicity. Immunohistochemistry, apoptosis assay, and auditory brainstem response (ABR) were employed to determine the impacts of RG108 on cisplatin-induced injury in murine hair cells (HCs) and spiral ganglion neurons (SGNs). Rhodamine 123 and TMRM were utilized for mitochondrial membrane potential (MMP) assessment. Reactive oxygen species (ROS) amounts were evaluated by Cellrox green and Mitosox-red probes. Mitochondrial respiratory function evaluation was performed by determining oxygen consumption rates (OCRs). The results showed that RG108 can markedly reduce cisplatin induced damage in HCs and SGNs, and alleviate apoptotic rate by protecting mitochondrial function through preventing ROS accumulation. Furthermore, RG108 upregulated BCL-2 and downregulated APAF1, BAX, and BAD in HEI-OC1 cells, and triggered the PI3K/AKT pathway. Decreased expression of low-density lipoprotein receptor-related protein 1 (LRP1) and high methylation of the LRP1 promoter were observed after cisplatin treatment. RG108 treatment can increase LRP1 expression and decrease LRP1 promoter methylation. In conclusion, RG108 might represent a new potential agent for preventing hearing loss induced by cisplatin via activating the LRP1-PI3K/AKT pathway.
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Objective To construct an evidence-based nursing plan for stroke patients with dysphagia for nursing by searching guidelines and using Delphi method. Methods The evidence-based nursing plan for stroke patients with dysphagia was constructed by literature review and guideline search. 15 experts from neurology, rehabilitation and nursing were selected to participate in two rounds Delphi consultation. Results The two-round response rates were 15/15. The expert authority coefficient was 0.877. The coordination coefficients were 0.180, 0.132 and 0.185 (P＜0.05). The final plan included 4 first-grade indicators, 14 second-grade indicators and 20 third-grade indicators. Conclusion The evidence-based nursing intervention plan for stroke patients with dysphagia based on Delphi method is reliable, which can provide reference for the rehabilitation nursing of stroke patients with dysphagia.
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Objective@#To construct an evidence-based nursing plan for stroke patients with dysphagia for nursing by searching guidelines and using Delphi method.@*Methods@#The evidence-based nursing plan for stroke patients with dysphagia was constructed by literature review and guideline search. 15 experts from neurology, rehabilitation and nursing were selected to participate in two rounds Delphi consultation.@*Results@#The two-round response rates were 15/15. The expert authority coefficient was 0.877. The coordination coefficients were 0.180, 0.132 and 0.185 (P<0.05). The final plan included 4 first-grade indicators, 14 second-grade indicators and 20 third-grade indicators.@*Conclusion@#The evidence-based nursing intervention plan for stroke patients with dysphagia based on Delphi method is reliable, which can provide reference for the rehabilitation nursing of stroke patients with dysphagia.
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Objective This study aims to examine the development of the posterior lateral line of the zebrafish and establish ant model to study the process of hair cell differentiation and regeneration .Methods We observed the posterior lateral line system formation by DAPI immunohistochemistry and whole mount in situ hybridization .We further evaluated hair cells differentiation within neuromast by using Transgenic Tg (Brn3c:mGFP) zebrafish and stained the functional hair cells by the mechanotransduction marker FM 1 -43FX .We labelled proliferating cells in primordium and neuromast by addition of BrdU to the system water .Results The posterior lateral line primordium originated from a sensory placode and started its journey at around 20 hours post fertilization to migrate along the horizontal myoseptum to the tail -tip with a constant speed (1 .7somite/hour) .The primordium depositd five or six neuromasts spaced along the body ,and two or three terminal neuromasts at the tail -tip at 48 hours post fertiliza-tion .At 3 ,5 and 7 days post fertilization ,zebrafish contained 5 .68 ± 1 .46 ,10 .1 ± 0 .99 ,and 12 .45 ± 1 .32 hair cells per neuromast ,respectively .Furthermore ,the average number of FM1-43FX stained hair cells within each neuro-mast were 3 .68 ± 1 .11 ,8 .18 ± 1 .86 ,and 10 .22 ± 1 .24 ,respectively .Conclusion We establish the development model of hair cells in zebrafish lateral line neuromast and suggest that 3 to 7 days post fertilization is an important period for lateral line neuromast differentiation .This study would be useful for underlying the mechanisms of hair cell differentiation and regeneration .
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Histone deacetylases are involved in many biological processes and have roles in regulating cell behaviors such as cell cycle entry, cell proliferation and apoptosis. However, the effect of histone deacetylases on the development of hair cells (HCs) has not been fully elucidated. In this study, we examined the influence of histone deacetylases on the early development of neuromasts in the lateral line of zebrafish. Hair cell development was evaluated by fluorescent immunostaining in the absence or presence of histone deacetylase inhibitors. Our results suggested that pharmacological inhibition of histone deacetylases with inhibitors, including trichostatin A, valproic acid and MS-275, reduced the numbers of both HCs and supporting cells in neuromasts. We also found that the treatment of zebrafish larvae with inhibitors caused accumulation of histone acetylation and suppressed proliferation of neuromast cells. Real-time PCR results showed that the expression of both p21 and p27 mRNA was increased following trichostatin A treatment and the increase in p53 mRNA was modest under the same conditions. However, the expression of p53 mRNA was significantly increased by treatment with a high concentration of trichostatin A. A high concentration of trichostatin A also led to increased cell death in neuromasts as detected in a TUNEL assay. Moreover, the nuclei of most of these pyknotic cells were immunohistochemically positive for cleaved caspase-3. These results suggest that histone deacetylase activity is involved in lateral line development in the zebrafish and might have a role in neuromast formation by altering cell proliferation through the expression of cell cycle regulatory proteins.