Research on the Anti-Aging Effects of Ganoderic Acid A
The continuous intensification of the global aging population trend poses severe challenges to the construction of the elderly health maintenance system and the control of medical costs. During the aging process, whether caused by age growth or external inducements, the body not only exhibits a decline in physiological functions but also presents characteristics of systemic regression with interactive effects across multiple systems. Therefore, preventing bodily aging from the source has become an important strategy to address the profound changes in population structure. Cellular senescence, as a key mechanism driving the deterioration of bodily functions and the occurrence of age-related diseases, has emerged as a crucial target in current anti-aging research. Among the relevant approaches, senolytic drugs centered on targeted clearance of senescent cells have attracted extensive attention. However, the effectiveness of this strategy is often limited by cell type specificity, and potential adverse reactions may occur while eliminating senescent cells. Based on a natural product library, this study established a multi-dimensional cellular screening system starting from anti-aging targets, providing a systematic platform for the identification of anti-aging active substances. Using this system, the study successfully screened out ganoderic acid A (GAA), verified its remarkable effects in delaying aging and improving tissue and metabolic functions through various models, and confirmed its in vivo safety, laying a foundation for the subsequent application of GAA. Further, with the help of human proteome microarray and other technologies, the study found that GAA delays cellular senescence by binding to the TCOF1 protein to regulate ribosomal homeostasis, thus revealing its mechanism of action.
To efficiently identify natural products with both safety and anti-aging activity, this study constructed a high-content evaluation system based on a multi-round screening strategy, aiming to systematically screen candidate molecules with anti-aging potential from 805 natural compounds. The first round of primary screening: Using the IMR-90 cell replicative senescence model combined with the PerkinElmer high-content imaging system, fully automated scanning of 45 fields of view was performed for cells in each well, and multiple indicators such as cell number, nuclear area, cell morphology and SA-β-Gal positive rate were comprehensively analyzed. The primary screening identified 112 compounds from 805 that could significantly reduce the proportion of SA-β-Gal positive cells, among which 47 had an inhibition rate exceeding 30%. The second round of secondary screening: Four senescence models of human umbilical vein endothelial cells (HUVEC) were constructed, including replicative senescence, H₂O₂-induced oxidative stress senescence, etoposide-induced genotoxic stress senescence and their combined stress model. The broad-spectrum anti-aging effects of candidate compounds were evaluated by detecting indicators such as LDH release and cell number. A total of 12 compounds with stable activity were obtained in this round of screening. The third round of final screening: Further experiments with gradient concentrations (0.1–100 μM) were carried out in the senescence models of mouse embryonic fibroblasts (MEF) and human liver L02 cells to comprehensively evaluate the anti-aging ability and cytotoxicity of candidate compounds. Finally, ganoderic acid A (GAA) was screened out, which exhibited significant anti-aging activity over a wide concentration range and had significantly lower toxicity than other candidates. This screening strategy constructed a senescence evaluation system covering multiple cell types and multiple induction methods, and introduced an intelligent image analysis algorithm to realize simultaneous detection of multiple parameters at the single-cell level and accurate identification of senescence characteristics (Figure 1).
Figure 1 Screening of anti-cellular senescence active compounds based on high-content analysis.
To comprehensively evaluate the anti-aging effects and safety of ganoderic acid A (GAA), the research team found through a cross-species, multi-model verification system that GAA could generally reduce the proportion of SA-β-Gal positive cells by 5% to 65% in 10 natural senescence cell models induced by different methods; in the Caenorhabditis elegans model, GAA dose-dependently extended the median lifespan (up to an 8% increase) and maximum lifespan (up to a 26% extension); in radiation-induced premature aging mice, GAA exhibited anti-aging ability comparable to that of dasatinib combined with quercetin; after 6 months of intervention in 16-month-old naturally aging mice, the expected lifespan of the mice at 24 months of age was extended by approximately 13 days (equivalent to an increase of more than 1 year in the expected lifespan of humans at the age of 69), the frailty index was reduced by 20%, and the senescence markers p53, p21 and p16 were significantly down-regulated; in high-fat diet-induced obese mice, GAA not only reduced the accumulation of senescent cells in organs such as the heart, liver and kidney, but also improved bone loss and muscle strength decline. Safety assessment showed that long-term administration did not cause abnormal liver and kidney functions in middle-aged mice, nor did it activate the proliferation of young cells or cancer cells (Figure 2).
Figure 2 Ameliorative effects of GAA intervention on the aging phenotype in aged mice.
To deeply reveal the mechanism of action of ganoderic acid A (GAA), the research team found through cross-model and cross-tissue proteomic analysis that GAA could significantly improve the decline in ribosomal function during aging. Functional experiments further confirmed that GAA could restore the impaired protein translation activity in senescent cells without overactivating the translation function of normal cells. Combined with rRNA transcription inhibitor (CX-5461), translation blocker (CHX) and gene silencing experiments, the study confirmed that GAA exerts its effect of delaying cellular senescence by regulating ribosome biogenesis and translation function to maintain ribosomal homeostasis, and this regulatory mechanism has been verified in various cell types (Figure 3). This study revealed the molecular mechanism by which GAA delays aging through the "ribosomal homeostasis regulation" pathway, providing a new theoretical basis and potential targets for the development of precision anti-aging drugs targeting ribosomal function.
Figure 3 Mechanism by which GAA delays cellular senescence through the regulation of ribosomal homeostasis.
To clarify the direct target of ganoderic acid A (GAA), the research team used HuProt™ 20K human protein microarray technology to screen more than 20,000 human proteins and found 345 proteins as potential binding targets of GAA, among which the ribosome biogenesis-related protein TCOF1 had a binding signal as high as 1.91, standing out significantly. Molecular docking analysis showed that the binding energy between GAA and TCOF1 was -5.8 kcal/mol, and the cellular thermal shift assay (CETSA) further verified their direct interaction. Functional studies showed that knockdown of TCOF1 could lead to the loss of approximately 90% of ribosomal function in senescent cells and completely eliminate the anti-aging effect of GAA. Notably, GAA does not alter the protein expression level of TCOF1, but exerts its effect by maintaining its phosphorylated state. DARTS experiments confirmed that GAA can both protect TCOF1 from protease degradation and prevent phosphatase-induced dephosphorylation. In summary, GAA effectively delays cellular senescence by stabilizing phosphorylated TCOF1 to maintain ribosomal homeostasis (Figure 4).
Figure 4 Molecular mechanism by which GAA targets TCOF1 to maintain ribosomal homeostasis and thereby inhibit cellular senescence.
This study screened and identified the key anti-aging molecule GAA from 805 natural products, and revealed a new mechanism by which it delays aging by binding to TCOF1 to regulate ribosomal homeostasis. The significant advantages of GAA in terms of broad-spectrum activity, safety and improvement of healthspan distinguish it from existing anti-aging drugs, opening up a new direction for health management against the backdrop of aging and providing critical data support.
Reference: Chen L, Wu B, Mo L, et al. High-content screening identifies ganoderic acid A as a senotherapeutic to prevent cellular senescence and extend healthspan in preclinical models[J]. Nat Commun, 2025, 16(1): 2878.
