104 A new study by a team of researchers from South-Central Minzu University and Guangxi University of Chinese Medicine has revealed the potential of a new composition of ursolic acid derivatives from Ludwigia hyssopifolia (LH) in inducing apoptosis in throat cancer cells. The findings, recently published in the Journal of Ethnopharmacology, highlight the effectiveness of LH-CUAD (Ludwigia hyssopifolia composition of ursolic acid derivatives) in targeting throat cancer through multiple cellular pathways. Ludwigia hyssopifolia, a traditional herb used in Guangxi Zhuang medicine, has long been employed for its therapeutic properties in treating throat disorders. The study aimed to isolate and evaluate the specific impacts of ursolic acid derivatives from LH on throat cancer cells. Previous research had indicated the anti-cancer properties of ethyl acetate and petroleum ether fractions of LH extracts, but the precise mechanisms remained unclear. The research team utilised solvent extraction, fractionation, chromatography, and semi-preparative high-performance liquid chromatography to extract and purify the LH-CUAD. The anti-throat cancer effects were then assessed using in vitro and in vivo models, specifically Hep-2 and FaDu throat cancer cell lines and Hep-2 tumour-bearing nude mice . The results were compelling. LH-CUAD significantly inhibited the proliferation and migration of throat cancer cells while exhibiting minimal toxicity to normal cells. Morphological changes indicative of apoptosis, such as cellular rounding and wrinkling, were observed under light microscopy. Further assays, including Hoechst 33258 staining and Annexin V-FITC/PI double-staining, confirmed that LH-CUAD induced apoptosis from early to late stages . The study delved deeper into the mechanisms by which LH-CUAD induces apoptosis. Key findings revealed that LH-CUAD activates the mitochondrial apoptotic pathway, inhibits the Akt/mTOR signalling pathway, and induces endoplasmic reticulum stress (ERS), all of which contribute to the apoptotic process in throat cancer cells . Mitochondrial pathway activation. LH-CUAD treatment led to the activation of a caspase cascade, a series of proteolytic enzymes playing a vital role in apoptosis. Western blotting showed increased levels of cleaved Caspase-7 and Caspase-12, markers of apoptosis, indicating the effectiveness of LH-CUAD in triggering cell death through mitochondrial dysfunction. Akt/mTOR pathway inhibition. The Akt/mTOR pathway is crucial for cell survival and proliferation. The study found that LH-CUAD significantly inhibited the phosphorylation of Akt and mTOR proteins, thereby blocking the pathway and promoting apoptosis. This inhibition was dose-dependent, with higher concentrations of LH-CUAD resulting in more substantial reductions in Akt and mTOR activity. Endoplasmic reticulum stress. Severe ERS can lead to apoptosis, and LH-CuAD was observed to trigger this stress in throat cancer cells. The study reported a decrease in the expression of GRP78, an ER chaperone protein, alongside increased levels of cleaved Caspase-7 and Caspase-12, highlighting the role of ERS in the apoptotic process induced by LH-CUAD . The antitumour activity of LH-CUAD was also evaluated in vivo using a xenograft model with nude mice. The results were promising, showing significant tumour growth suppression and low toxicity. Histological analyses further confirmed that LH-CUAD induced apoptosis in tumour tissues, mirroring the in vitro findings.