Yiqi Wenyang Jiedu prescription promotes disulfidptosis of gastric cancer by reversing senescent cancer-associated fibroblasts

Figure 1 Culture of MRC-5 cells and establishment of the cancer-associated fibroblasts model. A: Morphological characteristics of MRC-5 cells and cancer-associated fibroblasts (CAFs); B: Immunofluorescence detection of fibroblast activation protein in MRC-5 cells and CAFs; C: Immunofluorescence detection of α-smooth muscle actin in MRC-5 cells and CAFs. ^bP < 0.01. Error bars represent the standard error of the data. CAF: Cancer-associated fibroblasts; FAP: Fibroblast activation protein; α-SMA: α-smooth muscle actin.

Figure 2 Establishment of the senescent cancer-associated fibroblasts model and intervention effect of Yiqi Wenyang Jiedu prescription. A: Proportion of β-galactosidase staining-positive cells in the cancer-associated fibroblasts (CAFs) model and senescent CAFs (senCAFs) model under the same scale bar; B: Reversal effect on senCAFs by Yiqi Wenyang Jiedu prescription (YWJP)-containing serum at different concentrations vs blank serum (BS); C: Secretion levels of interleukin-6 (IL-6), interleukin-8 (IL-8), hyaluronic acid (HA), and HAase in senCAFs treated with YWJP at different concentrations vs BS, detected by ELISA; D: MRNA expression levels of IL-6, IL-8, HA, and HAase in senCAFs treated with YWJP at different concentrations vs BS, detected by quantitative PCR. ^aP < 0.05, ^bP < 0.01, NS: Not significant. Error bars represent the standard error of the data. CAF: Cancer-associated fibroblasts; YWJP: Yiqi Wenyang Jiedu prescription; BS: Blank serum; senCAFs: Senescent cancer-associated fibroblasts; IL-6: Interleukin-6; IL-8: Interleukin-8; HA: Hyaluronic acid.

Figure 3 Effect of Yiqi Wenyang Jiedu prescription on malignant behaviors of cells under HAase action. A: Effect of Yiqi Wenyang Jiedu prescription (YWJP) on migration of MKN45 cells in HAase environment; B: Effect of YWJP on colony-forming ability of MKN45 cells in HAase environment. ^bP < 0.01. Error bars represent the standard error of the data. YWJP: Yiqi Wenyang Jiedu prescription; BS: Blank serum.

Figure 4 Mechanism study on Yiqi Wenyang Jiedu prescription intervening in disulfidptosis of MKN45 gastric cancer cells. A: Effect of Yiqi Wenyang Jiedu prescription (YWJP) on the expression of SLC7A11 and GLUT1 in MKN45 cells in HAase environment; B: Effect of YWJP on the NADP⁺/NADPH ratio in MKN45 cells in HAase environment; C: Effect of YWJP on the expression of CYFIP1, Rac1, ARP2, and ARP3 in MKN45 cells in HAase environment; D: Effect of YWJP on the disulfide bond level in MKN45 cells in HAase environment; E: Effect of YWJP on PI staining of MKN45 cells. ^aP < 0.05, ^bP < 0.01, NS: Not significant. Error bars represent the standard error of the data. CAF: Cancer-associated fibroblasts; YWJP: Yiqi Wenyang Jiedu prescription; BS: Blank serum.