A novel combination therapy targets sonic hedgehog signaling by the dual inhibition of HMG-CoA reductase and HSP90 in rats with non-alcoholic steatohepatitis.

• Hedgehog signaling has been recognized as potentially promoting NASH. • Cholesterol affects hedgehog signaling by modifying PTCH1 and SMO activity. • HSP90 plays a role in the stability of SMO and GLI proteins. • Lovastatin and PU-H71 combined therapy effectively suppressed hedgehog signaling. • Lovastatin and PU-H71 improved biochemical parameters in rats with NASH. Non-alcoholic steatohepatitis (NASH) is characterized by liver inflammation, fat accumulation, and collagen deposition. Due to the limited availability of effective treatments, there is a pressing need to develop innovative strategies. Given the complex nature of the disease, employing combination approaches is essential. Hedgehog signaling has been recognized as potentially promoting NASH, and cholesterol can influence this signaling by modifying the conformation of PTCH1 and SMO activity. HSP90 plays a role in the stability of SMO and GLI proteins. We revealed significant positive correlations between Hedgehog signaling proteins (Shh, SMO, GLI1, and GLI2) and both cholesterol and HSP90 levels. Herein, we investigated the novel combination of the cholesterol-lowering agent lovastatin and the HSP90 inhibitor PU-H71 in vitro and in vivo. The combination demonstrated a synergy score of 15.09 and an MSA score of 22.85, as estimated by the ZIP synergy model based on growth inhibition rates in HepG2 cells. In a NASH rat model induced by thioacetamide and a high-fat diet, this combination therapy extended survival, improved liver function and histology, and enhanced antioxidant defense. Additionally, the combination exhibited anti-inflammatory and anti-fibrotic potential by influencing the levels of TNF-α, TGF-β, TIMP-1, and PDGF-BB. This effect was evident in the suppression of the Col1a1 gene expression and the levels of hydroxyproline and α-SMA. These favorable outcomes may be attributed to the combination's potential to inhibit key Hedgehog signaling molecules. In conclusion, exploring the applicability of this combination contributes to a more comprehensive understanding and improved management of NASH and other fibrotic disorders. The cholesterol-lowering drug lovastatin can disrupt Shh signaling by reducing cholesterol biosynthesis and influencing interaction between PTCH and Shh. PU-H71, an HSP90 inhibitor, could offer promise by inhibiting the chaperone machinery and impacting key proteins within the Shh pathway such as SMO and GLI1. Ch, cholesterol; GLI1, glioma-associated oncogene family zinc finger 1; HSP90, heat shock protein 90; PTCH, patched; Shh, sonic hedgehog; SMO, smoothened; SUFU, suppressor of fused. [Display omitted] [ABSTRACT FROM AUTHOR]