Boling Huazhuo Pill-medicated serum alleviates inflammation of RWPE-1 cells by regulating M1 macrophage-derived CCL3

Introduction

Chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) has a complex etiology and remains challenging to treat. Clinical studies have indicated that Boling Huazhuo Pill (BHP) is effective in managing CP/CPPS; however, its underlying mechanism remains unknown. Since increased macrophages and C-C motif chemokine ligand 3 (CCL3) release are closely associated with CP/CPPS pathogenesis, this study examined whether BHP-medicated serum reduces inflammation in RWPE-1 cells by regulating M1 macrophage-derived CCL3.

Methods

M0 and M1 macrophages were generated through THP-1 cell differentiation. Following treatment with BHP-medicated serum, macrophage polarization and inflammatory cytokine levels were analyzed using flow cytometry and enzyme-linked immunosorbent assay, respectively. The effects of BHP-medicated serum, alone or in combination with recombinant CCL3, on CCL3 production by M1 macrophages were investigated. Furthermore, the influence on proliferation, apoptosis, and nuclear factor kappa B (NF-κB) p65 activation in RWPE-1 cells was examined within a macrophage-RWPE-1 co-culture system.

Results

BHP-medicated serum reduced the proportion of M1 macrophages and suppressed inflammation. In the co-culture system, M1 macrophages inhibited RWPE-1 cell proliferation and induced apoptosis, whereas BHP-medicated serum reversed these effects. Notably, BHP-medicated serum reduced M1 macrophage-derived CCL3 production and inhibited NF-κB pathway activation in RWPE-1 cells. However, treatment with CCL3 counteracted these effects of BHP-medicated serum.

Conclusion

BHP-medicated serum may suppress NF-κB pathway activation by inhibiting M1 macrophage-derived CCL3, thereby reducing inflammation in RWPE-1 cells. These findings provide a theoretical basis for using BHP in CP/CPPS treatment.