ABSTRACT: This secondary integrative analysis of a previously published cohort investigated the molecular pathways underlying muscle recovery of the biceps femoris long head (BFLH) after a football match, by integrating newly generated transcriptomic data with clinical recovery markers over a 72-hour recovery period. BFLH muscle biopsies from 10 male, regional-first division football players were obtained seven days before a football match (MD-7) and three days post-match (MD+3). The final sample included 9 participants because 1 biopsy provided insufficient material for sequencing. Bulk RNA-sequencing was conducted to analyze differential gene expression considering Benjamini-Hochberg correction. Pathway enrichment analyses were conducted to identify biological processes altered between pre- and post-match conditions. Clinical markers, including creatine kinase (CK), hamstring rate of torque development (RTD), and sprint performance, were assessed before the match (MD) and at 24 (MD+1), 48 (MD+2), and 72 hours (MD+3) post-match. Correlations between transcriptomic and clinical variables were calculated using Spearman’s rank correlation. MYH1 was the most responsive gene, showing significant downregulation at MD+3 (log2FC = -1.116, q-value = 3.8 × 10-2). Pathway analyses identified 440 transcriptomic signatures at MD+3, reflecting stress response, immune and cytokine signaling, protein turnover, and structural remodeling. Changes in clinical variables during recovery were associated with coordinated expression shifts across genes related to different phases of the muscle damage-regeneration continuum, including muscle contraction, damage control, protein turnover, repair signaling, and myogenic regeneration. BFLH is not fully recovered three days after a football match, as evidenced by sustained downregulation of MYH1, pathway enrichment, and associations with clinical markers. Molecular recovery processes extend beyond 72 hours, suggesting this period is insufficient for complete BFLH recovery in football players.
