Theaflavin 3, 3-Digallate Delays Ovarian Aging by Improving Oocyte Quality and Regulating Granulosa Cell Function

Figure 6

TF3 may affect granulosa cell function by regulating autophagy through the mTOR pathway. pGCs were routinely cultured for 24 h and then starved in a serum-free culture medium for 24 h. TF3 (10 μM) or control treatment was given for 6 h. Cells were collected for transcriptome sequencing, and differentially expressed genes were subjected to GO analysis related to cell biological processes (a) and MCODE analysis related to signaling pathways (b). GSEA algorithm enrichment of all genes and calculation of enrichment scores were performed to find the key pathways affected by TF3 (c). Granulosa cells after TF3 treatment were collected, proteins were extracted, and the effect of TF3 on related proteins was analyzed by western blotting (d). Ovarian tissues were collected from all three groups, and proteins were extracted to analyze the effects of TF3 on NF-κB and mTOR protein levels (e). GAPDH was the internal reference protein. The molecular docking mode of TF3 to mTOR protein was predicted using AutoDock Vina software to map the action mode of TF3-5WBH, with the blue part indicating TF3 and the red box indicating the docking site (f).

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