Screening Of Chinese Herbal Extracts For Preventing Hair Cell Damage And Its Nano-Based System

With the rapid development of modern medical technology,people have deeply studied the deafness disease,so the prevention and treatment of deafness is coming to a new spring.At present,the treatment of deafness is using drug therapy,gene therapy,stem cell therapy,and so on.Natural medicine resources are rich in China.Traditional Chinese medicine is an important component of many drug libraries,and its research on the prevention and treatment of drug-induced hearing loss is also increasing.Nanoparticle delivery system can improve the bioavailability of drugs,which has attracted the attention of researchers in recent years.In modern clinical applications it has been shown that aminoglycosides are very effective antibacterial agents and that cisplatin is a chemotherapeutic agent commonly used for the treatment of cancer(such as lung cancer,prostate cancer,etc.),but it is well known that they cause ototoxicity,which greatly affected the patient’s quality of life.Therefore,while maintaining the efficacy of cisplatin or gentamicin,reducing their ototoxicity has been a problem that many researchers need to address.In this study,the HEI-OC1 cells and zebrafish were preliminarily modeled with gentamicin and cisplatin,and effective components were screened out from 224 Chinese herbal extracts.This laid the foundation for further discovery of compounds that can prevent ototoxicity.In this study,mouse cochlear hair cell line HEI-OC1 cells were used as a model.After experimental investigation,cells were exposed to the 10 mM gentamicin and 40 μM cisplatin for 24 h,respectively.40 μM EDA and 1 mM NAC were selected as positive agents for gentamicin and cisplatin,respectively.We have screened 224 Chinese herbal extracts and found that 13 of them could protected HEI-OC1 cell from gentamicin and cisplatin toxicity,and the cell survival rate is significantly increased.Using wild-type AB zebrafish lateral line as a model,after experimental investigation,the concentrations of gentamycin and cisplatin were 100 μM and 1 mM,respectively,and the exposure time was 1 h and 4 h,respectively.At the same time,80 μg/mL EDA and 40 μg/mL NAC were selected as the positive drugs for gentamicin and cisplatin,respectively.The experiment found that there are two kinds of traditional Chinese medicine extracts was treated as effective extracts,After the pretreatment of the effective extract,the degree of damage to the hair cells of the neuromast was significantly reduced.They protect zebrafish lateral line from gentamicin and cisplatin toxicity.Then it was explored whether the effective extracts screened in previous experiments will affect the antibacterial efficacy of gentamicin and the anti-cancer efficacy of cisplatin,and whether it will promote the regeneration of hair cells.The survival rate of non-small cell lung cancer A549 cells was measured by MTT assay,the antibacterial effect of gentamycin was determined by disk diffusion agar method tests,and the regeneration of zebrafish hair cells was examined by Brdu labeling method.It was found that 13 effective extracts did not affect the anticancer effect of cisplatin,nor did they affect the antibacterial effect of gentamicin.The effective extract was to protect hair cells from gentamicin damage,rather than to promote the rapid regeneration of hair cells.Finally,a preliminary study was conducted on the preparation of chitosan nanoparticles encapsulating effective extracts,and the uptake of Nile Red-loaded chitosan nanoparticles in zebrafish was examined.It was show that a large number of red fluorescent nanoparticles were taken into the body after 1h.Pharmacodynamic experiments showed that the chitosan nanoparticles loaded with effective extract can prevent HEI-OC1 cells and zebrafish from gentamicin or cisplatin damaging.In summary,this study indicated that the model of HEI-OC1 cells and zebrafish lateral line hair cells induced by gentamicin or cisplatin would be expected to be a new platform for the screening of anti-ototoxic drugs.The two selected extracts have preventive and therapeutic effects on ototoxicity induced by gentamicin or cisplatin.The CS NPs delivery system loaded with effective extract has certain potential application value in the field of deafness prevention and treatment.