Objective:To investigate whether asiaticoside(ASI)regulates the malignant biological behavior of hepatocellular carcinoma Huh7 cells via the cyclic adenosine monophosphate/protein kinase A/cAMP-response element binding protein(cAMP/PKA/CREB)signaling pathway.Methods:After determining the suitable ASI concentration and treatment duration using MTT assay,Huh7 cells were divided into the following groups:control,ASI-L(20 μmol/L),ASI-M(40 μmol/L),ASI-H(80 μmol/L),and ASI-H+Forskolin(80 μmol/L ASI+100 μmol/L cAMP activator Forskolin)groups.After 48 h of treatment,cell proliferation was assessed using MTT and colony formation assays;cell migration and invasion were analyzed using Transwell assays;apoptosis was detected using an Annexin V-FITC apoptosis detection kit.The secretion level of cAMP and the protein expression levels of phosphorylated PKA(p-PKA)and phosphorylated CREB(p-CREB)were evaluated using ELISA and Western blotting,respectively.A subcutaneous xenograft model was established by injecting Huh7 cells into the right abdomen of nude mice.ASI was administered by gavage at doses of 5,15,and 45 mg/kg for 4 weeks.Tumors were then harvested and weighed.Results:Treatment with 40 μmol/L ASI for 48 hours(close to the IC??)was determined to be the appropriate concentration and duration.Compared with the control group,the ASI-L,ASI-M,and ASI-H groups showed significantly reduced Huh 7 cell proliferation,colony formation,migration,invasion,cAMP levels,and expression of p-PKA/PKA and p-CREB/CREB(all P<0.05),while apoptosis rates were significantly increased(P<0.05).Compared with the ASI-H group,the ASI-H+Forskolin group exhibited significantly increased proliferation,colony formation,migration,invasion,cAMP level,and expression of p-PKA/PKA and p-CREB/CREB(all P<0.05),but apoptosis was significantly reduced(all P<0.05).In the nude mouse xenograft model,ASI at 5,15,and 45 mg/kg markedly decreased tumor weight in nude mice(all P<0.05).Conclusion:ASI inhibits the malignant biological behaviors and promotes apoptosis of Huh7 cells,as well as suppresses tumor growth in nude mice,by downregulating the expression of proteins in the cAMP/PKA/CREB signaling pathway.

Osteonecrosis of the femoral head （ONFH） is a painful and debilitating disease caused by impaired blood supply to the femoral head and cellular and tissue degeneration， leading to gradual destruction of the bone structure and progressive collapse of the femoral head. The main pathological mechanism of ONFH is the disruption of the balance between bone absorption and the reconstruction of new bone， resulting from microcirculation damage and decreased cellular tissue ability. This imbalance leads to biomechanical changes and accelerates the pathological progression of ONFH. In the early stages， clinical manifestations may not be obvious， mainly presenting as pain or discomfort in the hip or groin area， which can be relieved after rest. In the later stage of the disease， pain intensifies， and limb shortening， lower limb weakness， difficulty walking， or limping may occur. Currently， western medicine commonly uses osteogenic agents， anticoagulants， and artificial joint replacement for treatment， but there are also many issues such as prosthesis loosening and infection. Research has shown that traditional Chinese medicine （TCM） treatment of ONFH takes a holistic approach and employs multi-functional， multi-target， and multi-system Chinese medicine therapies， ensuring the safety and effectiveness of the treatment. The osteoprotegerin （OPG）/receptor activator of nuclear factor-κB （RANK）/RANK ligand （RANKL） signaling pathway plays a crucial role in maintaining the dynamic balance of bone remodeling. TCM treatments utilize this pathway to promote apoptosis of osteoclasts， reduce bone resorption， and accelerate bone formation， thereby playing an important role in the prevention and treatment of ONFH. This paper reviewed the role of OPG/RANK/RANKL signaling pathway and related cytokine expression in ONFH by reviewing relevant literature in China and abroad and research status of Chinese medicinal monomers， Chinese medicinal formulations， and combinations with physical therapy in increasing osteoblast secretion， promoting OPG expression， enhancing cytokine expression levels， and inhibiting osteoclast activity for the prevention and treatment of ONFH. This paper is expected to provide new ideas and directions for TCM in the prevention and treatment of ONFH.