Effects Of EGCG And TF On Brain Nerve Development Were Studied Based On The Brain Organoid Model

Epigallocatechin gallate(EGCG)and theaflavin(TF)are both bioactive components in tea with antioxidant,anticancer,and neuroprotective functions.In this study,specific differential genes were obtained with the help of the brain organoid model,transcriptome sequencing,and other technologies.The regulatory mechanisms of related signaling pathways were analyzed to explore the effects of bioactive tea components on neurodevelopment.The main findings are as follows.1.Construct brain organoids derived from human induced pluripotent stem cells(hi PSC)and explore the optimal induction time.In the culture of hi PSC-derived brain organoids,it was found that different induction times had significant differences in the morphology of brain organoids.Real-time fluorescence quantitative(RT-q PCR)and immunofluorescence(IF)methods were adopted.It was found that with the increase of induction time,the expression contents of excitatory,inhibitory,nerve barren cells and cortical specificity genes and proteins of brain organoids were down-regulated.The results suggested that the induction time of two days may be the best in the brain organoid culture.2.Transcriptomic sequencing technology was used to explore the molecular mechanism of EGCG expression and related genes’ response during brain organoid neurodevelopment.Firstly,30 Days of brain organoids were treated with 20 and 100 μM EGCG for 10 Days,and transcriptome gene sequencing was performed to screen the significantly differentially expressed transcriptome genes.Genomes using Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)Functional annotation and pathway analysis of differentially expressed transcriptome genes were performed.Sequencing results showed that during the neurodevelopmental process of brain organoids,223 gene expressions were up-regulated,and 217 gene expressions were down-regulated in the 100μM EGCG treatment group.Further,GO functional enrichment,KEGG pathway enrichment analysis,and expression verification of related differential gene RT-q PCR suggested that EGCG could affect the neural development of brain organoids by promoting the development and maturation of neuroimmune system and inhibiting the transmission of signals in synaptic structure.3.Transcriptomic sequencing technology was used to explore the molecular mechanism of TF expression and the response of related genes in the development of brain organoids.Firstly,30 Days of brain organoids were treated with 20 and 100 μM TF for 10 Days,and transcriptome genes were removed and sequenced to screen the significantly differentially expressed transcriptome genes.Functional annotation and pathway analysis of differentially expressed transcriptome genes were performed using GO and KEGG analysis.Sequencing results showed that68 gene expressions were up-regulated and 7 gene expressions down-regulated in the 20 μM TF treatment group,231 gene expressions up-regulated,and 35 gene expressions down-regulated in the 100 μM TF treatment group during the development of brain organoids.Further,GO functional enrichment analysis,KEGG pathway enrichment analysis,and RT-q PCR expression verification of related differential genes suggest that TF can affect the Wnt signaling pathway by promoting the development and maturation of neuron function and thus promote the neural development of the brain.