Regulation of monocyte polarization through nuclear factor Kappa B /inhibitor of Kappa B Alpha pathway by Cuscuta chinensis Lam. In postmenopausal osteoporosis.

ETHNOPHARMACOLOGICAL RELEVANCE

Cuscuta chinensis Lam. (CCL), is made from the dried mature seeds of a plant in the Convolvulaceae family. Predominantly distributed in China and several Asian countries, it has long been used to treat osteoporosis (OP) and other aging-related diseases. However, studies on the mechanisms of anti-OP compounds in CCL remain limited.

AIM OF THE STUDY

The objective of this study is to determine the bioactive constituents present in CCL and to elucidate their mechanisms of action in the prevention and treatment of postmenopausal osteoporosis (PMOP). This will be achieved by investigating the modulation of bone marrow macrophage polarization via the Nuclear Factor Kappa B (NF-κB)/Inhibitor of Kappa B Alpha (IκBα) signaling cascade.

MATERIALS AND METHODS

CCL's chemical components were identified using UPLC-Q-TOF-MS. Blood components were analyzed for targets using databases. Pathway enrichment was performed via network pharmacology. We used an ovariectomy (OVX)-induced OP rat model to assess the effects of CCL extracts in comparison to a positive control drug. Osteogenic markers were analyzed. We utilized flow cytometry to assess macrophage marker expression, while quantitative PCR (qPCR) and Western blotting were employed to identify targets within the signaling pathways.

RESULTS

Seventeen chemical components were identified in CCL extracts, of which 14 were identified as prototype compounds absorbed into the bloodstream. Pathway enrichment analysis revealed that CCL's therapeutic effects on PMOP were closely associated with the NF-κB signaling pathway, specifically targeting NF-κB and IκBα proteins. Animal studies showed that high-dose CCL significantly lowered serum levels of tartrate-resistant acid phosphatase (TRACP) and c-terminal telopeptide of type I collagen (CTX) (p < 0.01) and increased levels of bone-specific alkaline phosphatase (BALP) and procollagen I c-terminal propeptide (PICP) (p < 0.01), indicating effective inhibition of bone resorption and promotion of bone formation. CCL treatment improved the microstructure of trabecular bone at the distal femur by reducing bone cavity spaces, increasing trabecular thickness, and enhancing trabecular alignment. CCL markedly enhanced the expression levels of osteoprotegerin (OPG), runt-related transcription factor 2 (Runx2), and alkaline phosphatase (ALP) genes and their corresponding proteins in the tibial tissue (p < 0.01), promoting osteoblast differentiation and function. Flow cytometry analysis showed that CCL modulated immune cell markers CD86 and CD163, supporting its anti-inflammatory effects in PMOP treatment. Furthermore, CCL regulated the NF-κB/IκBα signaling pathway by significantly decreasing NF-κB expression and increasing IκBα expression, thereby modulating inflammatory responses and bone metabolism.

CONCLUSION

The active components in CCL effectively prevent and treat PMOP by modulating bone marrow macrophage polarization through the NF-κB/IκBα signaling pathway.