Risk signals are characteristic of many common inflammatory diseases and can function to activate nucleotide-binding oligomerization (NLR) family pyrin domain-containing 3 (NLRP3), the innate immune signal receptor in cytoplasm. The NLRP3 inflammasome plays an important role in the development of liver fibrosis. Activated NLRP3 nucleates the assembly of inflammasomes, leading to the secretion of interleukin (IL)-1β and IL-18, the activation of caspase-1, and the initiation of the inflammatory process. Therefore, it is essential to inhibit the activation of the NLRP3 inflammasome, which plays a vital role in the immune response and in initiating inflammation. RAW 264.7 and LX-2 cells were primed with lipopolysaccharide (LPS) for 4 h and subsequently stimulated for 30 min with 5 mM of adenosine 5′-triphosphate (ATP) to activate the NLRP3 inflammasome. Thymosin beta 4 (Tβ4) was supplemented to RAW264.7 and LX-2 cells 30 min before ATP was added. As a result, we investigated the effects of Tβ4 on the NLRP3 inflammasome. Tβ4 prevented LPS-induced NLRP3 priming by inhibiting NF-kB and JNK/p38 MAPK expression and the LPS and ATP-induced production of reactive oxygen species. Moreover, Tβ4 induced autophagy by controlling autophagy markers (LC3A/B and p62) through the inhibition of the PI3K/AKT/mTOR pathway. LPS combined with ATP significantly increased thee protein expression of inflammatory mediators and NLRP3 inflammasome markers. These events were remarkably suppressed by Tβ4. In conclusion, Tβ4 attenuated NLRP3 inflammasomes by inhibiting NLRP3 inflammasome-related proteins (NLRP3, ASC, IL-1β, and caspase-1). Our results indicate that Tβ4 attenuated the NLRP3 inflammasome through multiple signaling pathway regulations in macrophage and hepatic stellate cells. Therefore, based on the above findings, it is hypothesized that Tβ4 could be a potential inflammatory therapeutic agent targeting the NLRP3 inflammasome in hepatic fibrosis regulation.

Women going through menopause experience changes in their hormone levels, such as decreased estrogen secretion. Those changes can lead to weight gain and steatosis (fatty liver) due to abnormal lipid metabolism and bone turnover. Natural herbal medicines have been developed to treat and prevent menopausal symptoms. The aim of this study was to evaluate how a mixture of germinated Glycine max (GG) and Angelica gigas Nakai (AG) affected serum lipid profile levels, hepatic damage, inflammation, and bone turnover in ovariectomized (OVX) rats. The animals were randomly allocated into six groups: Sham control group (Sham), OVX control group (OVX), OVX + 50 mg/kg b.w. of GG (GG50) group, and OVX + 25, 50, 100 mg/kg b.w. of GG and AG mixture (GAM; GAM25, GAM50, GAM100) groups. After four weeks of treatment, the GAM groups exhibited decreases in serum lipid profile levels (TC, TG, and Low Density Lipoprotein (LDL)) and increases in High Density Lipoprotein (HDL). Proinflammatory mediators (IL-1β, IL-6, TNF-α, and iNOS) were reduced after the administration of GAM, and the degree of liver damage (ALT, AST) also decreased. Bone resorption (CTX1, NTX1, osteoclasts in H&E staining) decreased in the GAM-treated groups, and bone morphometric markers (BMD, BV/TV, Tb.N) improved compared with the OVX group. Additionally, increased bone formation (ALP, mineralization) and decreased bone resorption (TRACP) were confirmed in in vitro experiments. These results suggest that GAM has anti-obesity and anti-inflammatory effects by preventing dyslipidemia and that it alleviates bone mass loss in OVX rats by inhibiting osteoclastogenesis.