Objective:To explore the relationship between the expression of tumor necrosis factor receptor-associated factor 5 (TRAF5) and interferon regulatory factor 5 (IRF5) in intestinal mucosa of patients with ulcerative colitis and gut microbiota and intestinal mucosal barrier function.Methods:A total of 126 patients with ulcerative colitis in Qingdao Municipal Hospital from April 2021 to April 2023 were collected, according to the condition, there were 76 patients in the active phase and 50 patients in the remission phase, another 50 patients with intestinal polyps were taken as controls, intestinal mucosal tissues were collected from patients in three groups. Immunohistochemical staining was applied to detect the expression of TRAF5 and IRF5, the relationship between the expression of TRAF5 and IRF5 in intestinal mucosa of patients with ulcerative colitis and clinical symptoms, gut microbiota, and intestinal barrier function indicators was analyzed. Spearman method was applied for correlation analysis.Results:The positive expression rates of TRAF5 and IRF5 in intestinal mucosa of patients with active phase and remission phase ulcerative colitis were higher than those of patients with intestinal polyps: 85.53% (65/76) and 80.00% (40/50) vs. 40.00% (20/50), 81.58% (62/76) and 76.00% (38/50) vs. 50.00% (25/50), and the difference was statistically significant ( P<0.05); the expression of TRAF5 and IRF5 in intestinal mucosa of patients with ulcerative colitis was related to diarrhea, abdominal pain, purulent stool, tumor necrosis factor-α (TNF-α), Interleukin-6 (IL-6), C-reactive protein (CRP), erythrocyte sedimentation rate, Baron endoscopic score, inflammatory bowel disease questionnaire score, Mayo index score and Geboes index ( P<0.05). Compared with patients with negative expression of TRAF5 and IRF5, patients with positive expression of TRAF5 and IRF5 had fewer bifidobacteria and lactobacilli, and more enterobacteria and enterococcus: (8.72 ± 0.43) cfu/g vs. (6.85 ± 0.47) cfu/g, (9.74 ± 0.31) cfu/g vs. (8.26 ± 0.27) cfu/g, (9.73 ± 0.46) cfu/g vs. (11.06 ± 0.48) cfu/g, (7.64 ± 0.31) cfu/g vs. (8.47 ± 0.34) cfu/g; (8.82 ± 0.44) cfu/g vs. (6.73 ± 0.47) cfu/g, (9.13 ± 0.30) cfu/g vs. (8.22 ± 0.27) cfu/g, (11.09 ± 0.48) cfu/g vs. (9.87 ± 0.46) cfu/g, (7.76 ± 0.32) cfu/g vs. (8.48 ± 0.34) cfu/g, and the difference was statistically significant ( P<0.05). The levels of diamine oxidase, lipopolysaccharide and D-lactic acid in patients with positive expression of TRAF5 and IRF5 were obviously higher than those in patients with negative expression of TRAF5 and IRF5: (12.18 ± 2.75) mg/L vs. (7.56 ± 2.49) mg/L, (76.14 ± 13.86) ng/L vs. (37.57 ± 12.51) ng/L, (18.15 ± 4.83) U/L vs. (9.87 ± 3.25) U/L; (12.39 ± 2.72) mg/L vs. (7.65 ± 2.66) mg/L, (77.21 ± 13.79) ng/L vs. (40.87 ± 13.04) ng/L, (18.36 ± 4.75) U/L vs. (10.67 ± 3.86)U/L, and the difference was statistically significant ( P<0.05). The expression of TRAF5 in the intestinal mucosa of patients with ulcerative colitis was negatively correlated with the numbers of bifidobacteria and lactobacilli ( r = - 0.645 and - 0.646; P<0.05), and positively correlated with the number of enterobacteria, number of enterococcus, diamine oxidase, lipopolysaccharide and D-lactic acid ( r = 0.629, 0.589, 0.509, 0.606 and 0.596; P<0.05). The expression of IRF5 was negatively correlated with the numbers of bifidobacteria and lactobacilli ( r = - 0.701 and - 0.690; P<0.05), and positively correlated with the number of enterobacteria, number of enterococcus, diamine oxidase, lipopolysaccharide and D-lactic acid ( r = 0.690, 0.624, 0.605, 0.595 and 0.568; P<0.05). Conclusions:The positive rates of TRAF5 and IRF5 in intestinal mucosa of patients with ulcerative colitis is high, which is closely related to the imbalance of gut microbiota and the damage of intestinal mucosal barrier function.

Objective:To investigate the effects of oleanolic acid on the growth and migration of keloid fibroblasts.Methods:Keloid tissue samples from 9 patients in the Department of Plastic Surgery of General Hospital of Northern Theater were collected and fibroblasts were cultured in vitro. Fibroblasts were treated with different concentrations of oleanolic acid and divided into three groups: control group added 0.9% NaCl; 5 μmol/L oleanolic acid group added 5 μmol/L oleanolic acid; 10 μmol/L oleanolic acid group added 10 μmol/L oleanolic acid. MTT assay was used to detect cell proliferation; flow cytometry was used to detect cell cycle. Annexin V propidium iodide (AV-PI) staining was used to detect cell apoptosis. Transwell assay was used to detect the migration of oleanolic acid. Western blotting and real-time PCR were used to detect the expression of related proteins and mRNA activity. Each group was made in triplicate. Analysis of variance was used to compare the data among the three groups. LSD- t test was used for pairwise comparison, and P<0.05 was considered to be statistically significant. Results:MTT result showed that oleanolic acid could inhibit the proliferation of cells. After 24 hours, the proliferation of cells in 5 μmol/L oleanolic acid group and 10 μmol/L oleanolic acid group were 0.660±0.020 and 0.460±0.020, respectively, compared with 0.780±0.001 in the control group, F=114.4, P<0.001. Compared with the control group, the difference was statistically significant ( t=5.94, P<0.001, t=15.60, P<0.001); flow cytometry showed that the cell cycle G1/S phase transduction was blocked, 5 μmol/L oleanolic acid group and 10 μmol/L oleanolic acid group were significantly inhibited. The percentage of G1 phase cells in the 5 μmol/L oleanolic acid group was significantly higher than that in the control group ( t=3.14, P=0.030, t=6.38, P< 0.001). AⅤ-PI staining showed that the number of apoptotic cells in the 5 μmol/L oleanolic acid group (0.9%) and 10 μmol/L oleanolic acid group (3.4%) was significantly higher than that in the control group (0.4%), and the difference among the three groups was F=119.6, P<0.001. Transwell assay showed that the migration number of cells in 5 μmol/L oleanolic acid group (57.13 ± 2.65) and 10 μmol/L oleanolic acid group (42.15 ± 2.55) was significantly lower than that in control group (72.27± 3.32), F=101.3, P<0.001. Compared with the control group, the difference was statistically significant ( t=6.50, P<0.001, t=14.41, P<0.001). Western blotting showed that oleanolic acid could inhibit the expression of Cyclin D1, Bcl-2, Bax and MMP2. Compared with the control group, 5 μmol/L oleanolic acid t=8.70, P<0.001, t=5.00, P=0.040, t=12.41, P<0.001, t=10.46, P<0.001; compared with the control group, 10 μmol/L oleanolic acid t=31.61, P<0.001, t=23.17, P<0.001, t=12.11, P<0.001, t=44.52, P<0.001. Real-time PCR reaction showed that the mRNA activity levels of Cyclin D1, Bcl-2, Bax, MMP2 were also inhibited. Compared with the control group, 5 μmol/L oleanolic acid t=5.42, P< 0.001, t=3.11, P=0.040, t=16.11, P<0.001, t=11.71, P<0.001; compared with the control group, 10 μmol/L oleanolic acid t=51.78, P<0.001, t=30.89, P<0.001, t=10.64, P<0.001, t=17.10, P< 0.001. Conclusions:Oleanolic acid (5 μmol/L and 10 μmol/L) can inhibit the proliferation and migration of keloid fibroblasts and induce apoptosis of keloid fibroblasts after treating keloid fibroblasts for 24 hours, which can inhibit the growth of keloid and be used for the prevention and treatment of keloid.

Objective:To investigate the effects of oleanolic acid on the growth and migration of keloid fibroblasts.Methods:Keloid tissue samples from 9 patients in the Department of Plastic Surgery of General Hospital of Northern Theater were collected and fibroblasts were cultured in vitro. Fibroblasts were treated with different concentrations of oleanolic acid and divided into three groups: control group added 0.9% NaCl; 5 μmol/L oleanolic acid group added 5 μmol/L oleanolic acid; 10 μmol/L oleanolic acid group added 10 μmol/L oleanolic acid. MTT assay was used to detect cell proliferation; flow cytometry was used to detect cell cycle. Annexin V propidium iodide (AV-PI) staining was used to detect cell apoptosis. Transwell assay was used to detect the migration of oleanolic acid. Western blotting and real-time PCR were used to detect the expression of related proteins and mRNA activity. Each group was made in triplicate. Analysis of variance was used to compare the data among the three groups. LSD- t test was used for pairwise comparison, and P<0.05 was considered to be statistically significant. Results:MTT result showed that oleanolic acid could inhibit the proliferation of cells. After 24 hours, the proliferation of cells in 5 μmol/L oleanolic acid group and 10 μmol/L oleanolic acid group were 0.660±0.020 and 0.460±0.020, respectively, compared with 0.780±0.001 in the control group, F=114.4, P<0.001. Compared with the control group, the difference was statistically significant ( t=5.94, P<0.001, t=15.60, P<0.001); flow cytometry showed that the cell cycle G1/S phase transduction was blocked, 5 μmol/L oleanolic acid group and 10 μmol/L oleanolic acid group were significantly inhibited. The percentage of G1 phase cells in the 5 μmol/L oleanolic acid group was significantly higher than that in the control group ( t=3.14, P=0.030, t=6.38, P< 0.001). AⅤ-PI staining showed that the number of apoptotic cells in the 5 μmol/L oleanolic acid group (0.9%) and 10 μmol/L oleanolic acid group (3.4%) was significantly higher than that in the control group (0.4%), and the difference among the three groups was F=119.6, P<0.001. Transwell assay showed that the migration number of cells in 5 μmol/L oleanolic acid group (57.13 ± 2.65) and 10 μmol/L oleanolic acid group (42.15 ± 2.55) was significantly lower than that in control group (72.27± 3.32), F=101.3, P<0.001. Compared with the control group, the difference was statistically significant ( t=6.50, P<0.001, t=14.41, P<0.001). Western blotting showed that oleanolic acid could inhibit the expression of Cyclin D1, Bcl-2, Bax and MMP2. Compared with the control group, 5 μmol/L oleanolic acid t=8.70, P<0.001, t=5.00, P=0.040, t=12.41, P<0.001, t=10.46, P<0.001; compared with the control group, 10 μmol/L oleanolic acid t=31.61, P<0.001, t=23.17, P<0.001, t=12.11, P<0.001, t=44.52, P<0.001. Real-time PCR reaction showed that the mRNA activity levels of Cyclin D1, Bcl-2, Bax, MMP2 were also inhibited. Compared with the control group, 5 μmol/L oleanolic acid t=5.42, P< 0.001, t=3.11, P=0.040, t=16.11, P<0.001, t=11.71, P<0.001; compared with the control group, 10 μmol/L oleanolic acid t=51.78, P<0.001, t=30.89, P<0.001, t=10.64, P<0.001, t=17.10, P< 0.001. Conclusions:Oleanolic acid (5 μmol/L and 10 μmol/L) can inhibit the proliferation and migration of keloid fibroblasts and induce apoptosis of keloid fibroblasts after treating keloid fibroblasts for 24 hours, which can inhibit the growth of keloid and be used for the prevention and treatment of keloid.

Objective To assess the effect of platycodin D(PD)for alleviating pulmonary fibrosis in mice and explore the underlying mechanism.Methods C57BL/6J mouse models of pulmonary fibrosis induced by bleomycin injection into the airway were treated with daily intragastric administration of 10 mg/kg PD for 28 days.The changes of pulmonary fibrosis and the expression and distribution of transient receptor potential cation channel subfamily C member 6(TRPC6)were evaluated with immunohistochemistry,HE staining and Sirius Red staining.Western blotting was used to detect α-SMA expression in the lung tissues of the mice.Primary cultures of mouse lung fibroblasts were pretreated with PD(2.5,5.0,and 10 μmol/L)or larixyl acetate(LA;10 μmol/L)before exposure to 10 ng/mL transforming growth factor-β1(TGF-β1),and the changes in cell survival rate,expressions of collagen I,α-SMA and TRPC6,reactive oxygen species(ROS)production,mitochondrial membrane potential,and cell proliferation capacity were assessed.Network pharmacology analysis was performed to explore the mechanism by which PD alleviated pulmonary fibrosis.Results PD treatment significantly alleviated pulmonary fibrosis and reduced α-SMA expression in BLM-induced mouse models(P<0.05).In TGF-β1-induced primary mouse lung fibroblasts,PD effectively inhibited the cell proliferation,reduced ROS production(P<0.0001),rescued the reduction of mitochondrial membrane potential(P<0.001),and inhibited the expressions of α-SMA and collagenⅠ(P<0.05).Network pharmacology analysis suggested that TRPC6 mediated the effect of PD for alleviating pulmonary fibrosis.Immunohistochemistry showed that PD significantly reduced TRPC6 expression in the lung tissues of BLM-induced mice.In primary mouse lung fibroblasts,PD significantly inhibited TGF-β1-induced TRPC6 expression(P<0.05),and LA treatment obviously lowered the expression levels of TRPC6,α-SMA and collagenⅠ(P<0.05).Conclusion PD alleviated pulmonary fibrosis in mice possibly by down-regulating TRPC6 and reducing ROS production.

Objective To assess the effect of platycodin D(PD)for alleviating pulmonary fibrosis in mice and explore the underlying mechanism.Methods C57BL/6J mouse models of pulmonary fibrosis induced by bleomycin injection into the airway were treated with daily intragastric administration of 10 mg/kg PD for 28 days.The changes of pulmonary fibrosis and the expression and distribution of transient receptor potential cation channel subfamily C member 6(TRPC6)were evaluated with immunohistochemistry,HE staining and Sirius Red staining.Western blotting was used to detect α-SMA expression in the lung tissues of the mice.Primary cultures of mouse lung fibroblasts were pretreated with PD(2.5,5.0,and 10 μmol/L)or larixyl acetate(LA;10 μmol/L)before exposure to 10 ng/mL transforming growth factor-β1(TGF-β1),and the changes in cell survival rate,expressions of collagen I,α-SMA and TRPC6,reactive oxygen species(ROS)production,mitochondrial membrane potential,and cell proliferation capacity were assessed.Network pharmacology analysis was performed to explore the mechanism by which PD alleviated pulmonary fibrosis.Results PD treatment significantly alleviated pulmonary fibrosis and reduced α-SMA expression in BLM-induced mouse models(P<0.05).In TGF-β1-induced primary mouse lung fibroblasts,PD effectively inhibited the cell proliferation,reduced ROS production(P<0.0001),rescued the reduction of mitochondrial membrane potential(P<0.001),and inhibited the expressions of α-SMA and collagenⅠ(P<0.05).Network pharmacology analysis suggested that TRPC6 mediated the effect of PD for alleviating pulmonary fibrosis.Immunohistochemistry showed that PD significantly reduced TRPC6 expression in the lung tissues of BLM-induced mice.In primary mouse lung fibroblasts,PD significantly inhibited TGF-β1-induced TRPC6 expression(P<0.05),and LA treatment obviously lowered the expression levels of TRPC6,α-SMA and collagenⅠ(P<0.05).Conclusion PD alleviated pulmonary fibrosis in mice possibly by down-regulating TRPC6 and reducing ROS production.

Animals ; Humans ; Thymic Stromal Lymphopoietin ; Pyroglyphidae/metabolism* ; Cytokines/metabolism* ; Asthma/metabolism* ; Respiratory System ; Epithelial Cells/metabolism*