This study aimed to investigate the mechanism of the traditional Chinese medicinal formula Qijing Mingmu (QJMM) in the treatment of conjunctivochalasis (CCH) using network pharmacology and cellular experiments. Effective compounds and action targets of QJMM components were screened using the TCMSP and TCM-ID databases. Disease targets for CCH were obtained from the DisGeNET database, with gene names converted from the UniProt database. The Venny database yielded disease-drug intersection targets, which were input into the STRING database to generate protein-protein interaction (PPI) networks, followed by gene ontology (GO) and KEGG pathway analyses. Cytosccape software was employed to construct target networks for core components and therapeutic effects of QJMM, facilitating molecular docking. Conjunctival tissue was obtained from CCH and normal control groups (NC) (ethics approval number: PTEC-A-2024-30-1), subjected to immunohistochemical staining, and cultured as primary conjunctival fibroblasts. Fibroblasts were divided into NC, CCH, CCH+QJMM, and CCH+ nuclear factor κappa-B (NF-κB) inhibitor pyrrolidine dithiocarbamate (PDTC) groups. Immunofluorescence staining, cytokine assays, Western blot (WB), and quantitative reverse transcription PCR (qRT-PCR) were performed on each group. Results showed that 176 active compounds were screened from QJMM, yielding 293 target molecules in total. CCH identified 27 targets, with 12 overlapping targets identified via Venn diagram analysis. The PPI network diagram indicated tumor necrosis factor (TNF), matrix metalloproteinase (MMP)-2, MMP-9, and interleukin (IL)-6 as key targets. GO analysis yielded 91 enriched entries, while KEGG analysis identified 45 related entries and 110 signaling pathways, revealing the NF-κB signaling pathway. Molecular docking revealed that the core components of CCH exhibited strong binding affinity with the key target NF-κB, indicating potent biological activity. Subsequent immunohistochemical staining demonstrated significantly higher Toll-like receptor 4 (TLR4) and NF-κB expression in conjunctival tissues of the CCH group compared to the control group (P < 0.05). Conjunctival fibroblast experiments confirmed that NF-κB and its upstream factors TLR4 and myeloid differentiation primary response 88 (MYD88) exhibited higher protein and mRNA expression in the CCH group than in the NC group. Furthermore, expression of downstream inflammatory cytokines IL-6, IL-1β, and TNF-α was significantly elevated compared to the NC group (P < 0.05). Following administration of QJMM and PDTC, the expression of NF-κB and its upstream factors TLR4 and MYD88 protein, mRNA and immunofluorescence intensity decreased compared to the CCH group (P < 0.05). In conclusion, QJMM exerts multi-targeted, multi-pathway regulatory effects in treating CCH. Both network pharmacology and experimental studies confirm NF-κB plays a pivotal role in CCH pathogenesis and progression. QJMM effectively treats CCH by inhibiting the NF-κB signaling pathway.