Integrated network pharmacology and therapeutic verification of Dendrobium officinale flower on primary dysmenorrhea via inhibition of the TLR4/NF-κB/COX-2 pathway

Abstract

Dendrobium officinale flowers (DOF) offer a wide range of therapeutic benefits due to their rich content of bioactive compounds. Traditionally used in Chinese medicine, these flowers are valued for their health benefits, including anti-inflammatory and antioxidant properties, which may contribute to their therapeutic effects. The anti-inflammatory and antioxidant extract of DOF exhibited anti-primary dysmenorrhea (PD) activity in oxytocin-induced mice. This study aimed to unravel the underlying mechanisms of DOF-EA in treating PD using an integrative approach that combines network pharmacology and experimental verification. The active fraction, DOF-EA, was identified through anti-inflammatory and antioxidant activity assays. The phytochemical profile of DOF-EA was characterized by HPLC analysis. The therapeutic mechanisms of DOF-EA in treating PD were predicted using network pharmacology and molecular docking analyses. Moreover, the therapeutic effects of DOF-EA were confirmed using oxytocin-induced mice and uterine strip models. Enzyme activity assays and immunoblotting methods were employed for target validation. DOF-EA inhibited COX-2 with an IC50 of 0.44 μg/mL, scavenged DPPH radicals with an IC50 of 9.88 μg/mL, and suppressed NO production in RAW 264.7 cells with an IC50 of 8.08 μg/mL. Additionally, DOF-EA reduced the writhing response and decreased TNF-α and PGF_2α levels in PD mice. It also inhibited contraction activity in uterine strips by blocking calcium channels. Furthermore, DOF-EA downregulated the expression of TLR4, NF-κB, COX-2, and p-IκBα, while upregulating the expression of HO-1 and Nrf2 in the uterine tissue of PD mice. In conclusion, DOF-EA exerts therapeutic effects against PD by inhibiting TLR4/NF-κB/COX-2 and promoting the Nrf2/HO-1 pathway.