Autophagy-targeting modulation to promote peripheral nerve regeneration

Nerve regeneration following traumatic peripheral nerve injuries and neuropathies is a complex process modulated by diverse factors and intricate molecular mechanisms.Past studies have focused on factors that stimulate axonal outgrowth and myelin regeneration.However,recent studies have highlighted the pivotal role of autophagy in peripheral nerve regeneration,particularly in the context of traumatic injuries.Consequently,autophagy-targeting modulation has emerged as a promising therapeutic approach to enhancing peripheral nerve regeneration.Our current understanding suggests that activating autophagy facilitates the rapid clearance of damaged axons and myelin sheaths,thereby enhancing neuronal survival and mitigating injury-induced oxidative stress and inflammation.These actions collectively contribute to creating a favorable microenvironment for structural and functional nerve regeneration.A range of autophagyinducing drugs and interventions have demonstrated beneficial effects in alleviating peripheral neuropathy and promoting nerve regeneration in preclinical models of traumatic peripheral nerve injuries.This review delves into the regulation of autophagy in cell types involved in peripheral nerve regeneration,summarizing the potential drugs and interventions that can be harnessed to promote this process.We hope that our review will offer novel insights and perspectives on the exploitation of autophagy pathways in the treatment of peripheral nerve injuries and neuropathies.

Study on the molecular mechanism of rapamycin-induced autophagy in acute T lymphoblastic leukemia cells

Objective:To study the effect of different concentrations of rapamycin on the proliferation of acute leukemia CD4^(+)T-Jurkat cells,and to explore its mechanism from the aspect of autophagy.Methods:The effect of different concentrations of rapamycin on cell proliferation was detected by MTT assay;Apoptosis rate and cell cycle arrest were detected by flow cytometry;The changes of autophagic lysosomes were observed by acridine orange staining;Transcriptome sequencing data were used to analyze the differential expression of autophagy-related genes;The transcription and protein expression levels of autophagy-related genes were detected by RT-qPCR and Western Blot;The chromatin accessibility of the promoter region of autophagy gene was analyzed by FAIRE-qPCR.Results:Compared with the control group,rapamycin inhibited the proliferation of Jurkat cells in a concentration-and time-dependent manner.The cell cycle was arrested in G0/G1 phase,but it could not effectively induce apoptosis.Observed by acridine orange staining,the number of autophagic lysosomes increased after administration.RNA-seq data showed that rapamycin could significantly affect the transcription level of autophagy-related genes.After Jurkat cells were treated with different concentrations of rapamycin for 48 h,10 nM and 20 nM rapamycin could up-regulate the expression of ULK1,ATG13,ATG16L2,PI3K3R1,Raptor genes and down-regulate the expression of MAPK1 gene.At 50 nM,the expression levels of each gene were down-regulated.The chromatin accessibility of the autophagy gene promoter region also changed,which was basically consistent with the trend of gene expression.Conclusion:Rapamycin can inhibit the proliferation of Jurkat cells,block cell cycle and induce autophagy.Low concentrations of rapamycin promoted the expression of autophagy-related genes,while high concentrations inhibited their expression.

Functional Analysis of Autophagy Genes via Agrobacterium-Mediated Transformation in the Vascular Wilt Fungus Verticillium dahliae

Autophagy is a widely conserved intracellular process for degradation and recycling of proteins, organelles and cytoplasm in eukaryotic organisms and is now emerging as an important process in tbliar infection by many plant pathogenic fungi. However, the role of autophagy in soil-borne fungal physiology and infection biology is poorly understood. Here, we report the establishment of an Agro- bacterium tumefaciens-mediated transformation （ATMT） system and its application to investigate two autophagy genes, VdATG8 and VdATG12, by means of targeted gene replacement and complementation. Transformation of a cotton-infecting Verticillium dahliae strain Vd8 with a novel binary vector pCOM led to the production of 384 geneticin-resistant translbnnants per 1 × 10^4 conidia. V. dahliae mutants lacking either VdATG8 or VdATGI2 exhibited reduced conidiation and impaired aerial hyphae production. Disease development on Arabidopsis plants was slightly delayed when inoculated with VdATG8 or VdATG12 gene deletion mutants, compared with the wild- type and gene complemented strains. Surprisingly, in vitro inoculation with unimpaired roots revealed that the abilities of root invasion were not affected in gene deletion mutants. These results indicate that autophagy is necessary for aerial hyphae development and plant colonization but not for root infection in E dahliae.

Resistance genes mediate differential resistance to pine defensive substances α-Pinene and H_(2)O_(2) in Bursaphelenchus xylophilus with different levels of virulence

The pine wood nematode(PWN),Bursaphelenchus xylophilus(Steiner & Buhrer) Nickle,is the pathogen of pine wilt disease(PWD) which can devastate forests.PWN can be of hi gh or low severity and the mechanisms underlying the differences in virulence are unclear.Therefore,it is necessary to study the relationship between differentiation of PWN severity and its resistance to the main defensive substances of pine species(i.e.,α-pinene and H_(2)O_(2)).The feeding rate and fecundity of PWN was examined at different levels of virulence under conditions of a-pinene and H_(2)O_(2) stress.Moreover,the expression patterns of the main resistance genes of PWN with different virulence were determined under conditions of α-pinene and H_(2)O_(2) stress.The feeding rate and fecundity of the high virulence strain AMA3 were higher than those of the low virulence strain YW4.The expression levels of the autophagy gene BxATG5,cytochrome P450 gene BxCYP33 D3,and glutathione S-transferase genes BxGST1 and BxGST3 in AMA3 increased significantly upon exposure to α-pinene for 2 h,while these genes showed smaller degrees of upregulation in YW4.Under conditions of H_(2)O_(2) stress,the expression levels of BxATG5,catalase genes Bxy-ctl-1 and Bxy-ctl-2,and the 2-cysteine peroxiredoxin gene BxPrx in AMA3 were higher than those in YW4.These findings suggest that high virulence PWN has greater resistance to pine defensive substances α-pinene and H_(2)O_(2) than low virulence PWN,and resistance genes mediate the differential resistance of PWN strains.This study will contribute to the clarification of the mechanism underlying virulence differentiation of PWN and will advance understanding of the pathogenic mechanism of PWD.

A mechanism to improve early diabetic nephropathy by modulating autophagy-related mechanisms with Yuye Decotion

Objective: To investigate the molecular mechanisms of Yuye Decotion in the treatment of diabetic kidney disease using network pharmacology methods and molecular docking techniques. Methods: Obtain the transcriptome gene expression of diabetic nephropathy through GEO database, and extract genes related to autophagy. Screen the active ingredients and corresponding targets of Yuye Decoction through the TCMSP database, and map the drug prediction targets and disease targets to obtain the autophagy-related Yuye treatment targets for diabetic nephropathy point. Use String database combined with Cytoscape 3.7.2 software to construct the "drug-active ingredient-target" network and protein interaction network of Yuyetang for the treatment of diabetic nephropathy. The target point of liquid soup in the treatment of diabetic nephropathy was analyzed by GO biological process enrichment analysis and KEGG pathway enrichment analysis, and finally used Pymol and other software to analyze the core active components of Yuye Decotion and The core target protein undergoes molecular docking verification. Results: (i)100 eligible diabetic nephropathy and autophagy related genes were screened, and the potential targets of Yuye Decoction were 1,428. The acquired genes related to diabetic nephropathy and autophagy were mapped to potential targets of Yuye Decoction, and 22 therapeutic targets were obtained. GO biological process enrichment analysis and KEGG pathway enrichment analysis found that the pathways related to autophagy in the treatment of diabetic nephropathy by Yuye Decoction may include mTOR signaling pathway, phospholipase D signaling pathway, insulin resistance, EGFR tyrosine kinase inhibitor resistance, Apoptosis, PI3K /Akt signaling pathway, NF-κB signaling pathway, etc. (ii)The protein interaction network shows that VEGFA, ERBB2, GASP3, MAPK8, MYC, CDKN1A, EGFR, BCL2L1 may be the key targets of Yuye Decoction in the treatment of diabetic nephropathy. Molecular docking realizes the binding of 4 core active ingredients to 8 core target proteins. Conclusions: The research results show that Yuye Decoction treats diabetic nephropathy through multi-component, multi-target, and multi-pathway action, and provides new theoretical basis for the study of pharmacological effects and clinical application of Yuye Decoction in the treatment of diabetic nephropathy in autophagy-related aspects.