International Journal of Molecular Medical Science, 2024, Vol.14, No.6, 324-341 http://medscipublisher.com/index.php/ijmms 330 Polysaccharides from Actinidia eriantha roots (AEP) also exhibited significant antitumor activity in mouse models by promoting splenocyte proliferation, NK cell activity, and cytokine production, thereby enhancing the immune response (Xu et al., 2009). Additionally, polysaccharides fromPanax japonicus (PSPJ) repressed H22 tumor growth in vivo by modulating immune responses and reducing immunosuppressive factors in tumor-associated macrophages (Shu et al., 2018). Sanghuangporus polysaccharides exhibit potent antitumor effects through various mechanisms, including cell cycle regulation, apoptosis induction, and immune modulation. These findings highlight the potential of Sanghuangporus polysaccharides as natural therapeutic agents for cancer treatment. 4 Mechanisms of Sanghuangporus Polysaccharides-Induced Tumor Cell Apoptosis 4.1 Methods for detecting apoptosis To investigate the antitumor effects of Sanghuangporus polysaccharides, various methods are employed to detect apoptosis in cancer cells. Common techniques include the Cell Counting Kit-8 (CCK-8) assay, which measures cell viability, and Hoechst staining, which identifies apoptotic cells by their characteristic nuclear morphology (Figure 5) (Ding et al., 2020). Flow cytometry is another critical method used to analyze cell cycle distribution and quantify apoptotic cells by detecting sub-G1 peaks (Cao et al., 2010). Additionally, TUNEL assay and PI staining are utilized to confirm DNA fragmentation and other morphological changes associated with apoptosis (Song et al., 2014; Chen, 2024). Figure 5 Inhibitory effects of Sargassum fusiforme polysaccharide (SFPS) on the growth of human erythroleukemia (HEL) cells and human embryonic lung (MRC-5) cells relative to DMEM-treated control cultures (Adopted from Ding et al., 2020) Image caption: Data are presented as the mean ± SD (n=5). HEL cells and MRC-5 cells were incubated with (A) SFPS I, (B) SFPS II, and (C) SFPS III (10~100 μg·mL−1) for 24 h. (D) IC50 values for the inhibition of HEL cell viability by different purified components of SFPS. Cell viability was measured using CCK-8 and MTT assays (Adopted from Ding et al., 2020) Western blot analysis is frequently used to detect the expression of apoptosis-related proteins, providing insights into the molecular mechanisms underlying apoptosis (Cao et al., 2010; Bie et al., 2020). Real-time quantitative polymerase chain reaction (qPCR) is employed to measure the mRNA levels of genes involved in apoptosis, offering a comprehensive understanding of gene regulation during the apoptotic process (Ding et al., 2020; Xu et al., 2021). 4.2 Study of apoptotic signaling pathways Sanghuangporus polysaccharides induce apoptosis through multiple signaling pathways. One of the primary pathways is the intrinsic mitochondrial pathway, which involves the disruption of mitochondrial membrane potential and the release of cytochrome c into the cytoplasm. This pathway is characterized by the activation of
RkJQdWJsaXNoZXIy MjQ4ODYzNQ==