1.Therapeutic Effect and Mechanism of Regulating Inflammasome NLRP3 Signaling Pathway of Lycopi Herba Extract on Chronic Prostatitis
Yongjiao HUA ; Lina LIU ; Liang LI ; Yaozhong LYU ; Wei XIAO
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(15):51-59
ObjectiveTo investigate the therapeutic effect of Lycopi Herba extract on chronic prostatitis (CNP) and explore the underlying action mechanism via the inflammasome NOD-like receptor protein 3 (NLRP3) pathway. MethodNormal human prostatic stromal cells, namely WPMY-1 were induced by lipopolysaccharide (LPS) of 5 mg·L-1, and the effects of Lycopi Herba extract of 3.125, 6.25, 12.5, 25, 50, and 100 mg·L-1 on interleukin-6 (IL-6) level released by LPS-induced WPMY-1 cells were detected by enzyme-linked immunosorbent assay (ELISA). The half-maximal inhibitory concentration (IC50) was calculated. The expression of key proteins in the NLRP3 pathway was detected by western blot after Lycopi Herba extract of 50, 75, and 100 mg·L-1 was administered to WPMY-1 cells. The rat model of CNP was established by injecting carrageenan salt solution into the abdominal lobe of the prostate gland. Hematoxylin-eosin (HE) staining was used to observe the histopathological changes in the prostate gland in rats. The prostate organ index of rats was measured. The level of 5α-dihydrotestosterone (5α-DHT) in serum, as well as the levels of IL-6, tumor necrosis factor-α (TNF-α), transforming growth factor-β1 (TGF-β1), cyclooxygenase-2 (COX-2), prostaglandin E2 (PGE2), and inducible nitric oxide synthase (iNOS) in prostate tissue were detected by ELISA. The key protein expressions of COX-2, TGF-β1, and NLRP3 pathway in prostate tissue were detected by Western blot. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was used to detect the expressions of COX-2, IL-1β, TGF-β1, and TNF-α mRNA in prostate tissue. ResultCompared with the normal group, the level of IL-6 and the protein expression levels of NLRP3, ASC, Caspase-1, and IL-1β of WPMY-1 cells in the model group were increased (P<0.05, P<0.01). Compared with the model group, Lycopi Herba extract could inhibit the levels of IL-6 (P<0.01) released by LPS-induced WPMY-1 cells, with IC50 of 38.26 mg·L-1. The protein expression levels of NLRP3, ASC, and IL-1β in the low-, medium-, and high-dose groups of Lycopi Herba extract were significantly down-regulated (P<0.05, P<0.01). The expression levels of Caspase-1 protein in medium- and high-dose groups of Lycopi Herba extract were significantly down-regulated (P<0.05, P<0.01). Compared with the sham operation group, the prostate organ index of rats in the model group was significantly increased (P<0.01), a large number of inflammatory cells were infiltrated in the prostate tissue, and the histopathological score was significantly increased (P<0.05); the levels of 5α-DHT in serum, the levels of TNF-α, PGE2, IL-6, TGF-β1, NOS2/iNOS, and COX-2 in prostate tissue, and expression levels of COX-2, IL-1β, and TGF-β1 were significantly increased (P<0.05, P<0.01). The mRNA expression levels of COX-2, TGF-β1, NLRP3, Caspase-1, ASC, and IL-1β in prostate tissue were significantly up-regulated (P<0.05, P<0.01). Compared with model group, the low and high doses of Lycopi Herba extract could alleviate the pathological changes in prostate tissue induced by carrageenan, significantly reduce the level of 5α-DHT in serum, levels of TNF-α, PGE2, TGF-β1, and iNOS in prostate tissue (P<0.05, P<0.01), and mRNA expression levels of COX-2, IL-1β, and TGF-β1 (P<0.05, P<0.01). The protein expression levels of COX-2, Caspase-1, ASC, and NLRP3 in prostate tissue were significantly down-regulated (P<0.05, P<0.01). The prostate organ index of the low-dose group of Lycopi Herba extract was significantly decreased (P<0.01). The level of COX-2 in prostate tissue of the high-dose group of Lycopi Herba extract was significantly decreased, and the protein expression levels of TGF-β1 and IL-1β were significantly down-regulated (P<0.05). ConclusionLycopi Herba extract has an obvious therapeutic effect on CNP and may reduce inflammation by inhibiting the activation of the inflammasome NLRP3 signaling pathway.