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ObjectiveTo investigate the effect of modified Weijingtang on the pyroptosis of RAW264.7 macrophages via the cysteinyl aspartate-specific protease-1 (Caspase-1)/gasdermin D (GSDMD) pathway. MethodLipopolysaccharide (LPS) was used to induce pyroptosis of RAW264.7 cells. The blank group was treated with the blank serum, and the intervention groups were treated with the sera containing different doses of modified Weijingtang. After 24 h, the viability of cells in different groups was examined by the cell counting kit-8 (CCK-8). The pyroptosis and morphology of cells in each group were observed by a scanning electron microscope and a phase-contrast microscope, respectively. The mRNA and protein levels of nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3), Caspase-1, and GSDMD in each group were determined by real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) and Western blot, respectively. The levels of interleukin (IL)-18 and IL-1β in each group were measured by enzyme-linked immunosorbent assay. ResultUnder the electron microscope, RAW264.7 cells presented the best morphology and structure in the blank group and obvious pyroptosis and leakage of cell contents in the model (LPS) group. Compared with the model group, the intervention groups showed reduced pyroptosis to varying degrees, and the high-dose group had the closest cell morphology and structure to the blank group. Under the optical microscope, RAW264.7 cells were spherical in the blank group and irregular with protrusions in the model group. Compared with the model group, the intervention groups showed improved cell morphology, and the cell morphology in the group with the dose of 20% was the closest to that in the blank group. The mRNA and protein levels of NLRP3, Caspase-1, and GSDMD in the model group were higher than those in the blank group (P<0.05). Compared with the model group, each intervention group showed down-regulated expression of the above indicators (P<0.05). Compared with the blank group, the model group presented elevated levels of IL-18 and IL-1β (P<0.05), which were lowered in the intervention (10%, 20%) groups (P<0.01). ConclusionModified Weijingtang inhibits the pyroptosis of macrophages by down-regulating the Caspase-1/GSDMD pathway and reducing the release of proinflammatory cytokines.
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Cholestatic liver disease is a common disease of the hepatobiliary system. Its etiology and pathogenesis are complex. The establishment of an appropriate animal model of cholestatic liver disease is the basis for further study of its pathogenesis and prevention. This study summarized the existing modeling methods, mechanisms, and characteristics of this model, and analyzed its alignment with the clinical disease and syndrome characteristics of integrated traditional Chinese and Western medicine based on the modern clinical diagnostic criteria and traditional Chinese medicine syndrome characteristics of cholestatic liver disease, so as to provide a reference for establishing standard animal models and evaluation methods for cholestatic liver disease that accord better with the clinical practice of integrated traditional Chinese and Western medicine.
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OBJECTIVE@#To investigate the protective effect of procyanidin B2 (PCB2) on the intestinal barrier and against enteritis in mice with trinitrobenzene sulphonic acid (TNBS)-induced colitis and explore the possible mechanism.@*METHODS@#A mouse model of TNBS-induced colitis was established in male Balb/c mice aged 6-8 weeks. The successfully established mouse models were randomly divided into PCB2 treatment group (=10) and model group (=10) and were treated with daily intragastric administration of PCB2 (100 mg/kg, 0.2 mL) and 0.2 mL normal saline, respectively. After 4 weeks, the disease symptoms, intestinal inflammation, intestinal mucosal cell barrier function and the changes in PI3K/AKT signaling were evaluated using HE staining, immunofluorescence assay and Western blotting.@*RESULTS@#The disease activity index of the mice was significantly lower and the mean body weight was significantly greater in PCB2 group than in the model group in the 3rd and 4th weeks of intervention ( < 0.05). The levels of colonic inflammation and intestinal mucosal inflammatory mediators IL-1β and TNF-α were significantly lower while IL-10 was significantly higher in PCB2 group than in the model group ( < 0.05). Compared with those in the model group, the mice in PCB2 treatment group showed a significantly lower positive rate of bacterial translocation in the mesenteric lymph nodes and a lower thiocyanate-dextran permeability of the intestinal mucosa ( < 0.05). Western blotting showed that PCB2 treatment significantly increased the expressions of claudin-1 and ZO-1 ( < 0.05) and significantly lowered the expression levels of p-PI3K and p-AKT in the intestinal mucosa as compared with those in the model group ( < 0.05).@*CONCLUSIONS@#PCB2 suppresses intestinal inflammation and protects intestinal mucosal functions and structural integrity by inhibiting intestinal PI3K/AKT signaling pathway, suggesting the potential of PCB2 as a new drug for Crohn's disease.
Subject(s)
Animals , Male , Mice , Biflavonoids , Catechin , Colitis , Colon , Enteritis , Intestinal Mucosa , Phosphatidylinositol 3-Kinases , Proanthocyanidins , Trinitrobenzenesulfonic AcidABSTRACT
Objective To investigate the clinical efficacy of intraoperative fluorouracil implant combined with raltitrexed chemotherapy in advanced gastric cancer. Methods The clinical data of patients with advanced gastric cancer from November 2013 to November 2014 were retrospectively analyzed. The patients were divided into 2 groups according to intraoperative treatment method. Sixty-two cases (observation group) received intraoperative fluorouracil implant combined with raltitrexed regional chemotherapy, and 54 cases (control group) were not given the intraoperative chemotherapy drugs. The postoperative ventilation time, incidence of complications, peripheral blood white blood cell and platelets 1st, 3rd, 5th and 7th day after operation, cumulative recurrence rate and cumulative survival rate 3 years after operation were compared between 2 groups. Results There were no significant differences in postoperative ventilation time and incidence of complications between 2 groups (P > 0.05). The white blood cell 1st and 3rd day after operation in observation group was significantly lower than that in control group: (5.21 ± 1.03)×109/L vs. (6.52 ± 1.08)×109/L and (5.29 ± 1.11)×109/L vs. (6.37 ± 1.06)×109/L, the platelet 1st, 3rd and 5th day after operation in observation group was significantly lower than that in control group: (172.64 ± 31.48) × 109/L vs. (188.34 ± 30.05) × 109/L, (175.81 ± 31.77) × 109/L vs. (190.36 ± 31.12) ×109/L and (178.46 ± 32.04) ×109/L vs. (191.18 ± 31.29) ×109/L, and there were statistical differences (P<0.05); but the white blood cell and platelets in 2 groups were in the normal range at all time points. The 3-year cumulative recurrence rate in the observation group was significantly lower than that in control group: 75.8% (47/62) vs. 83.3% (45/54), the 3-year cumulative survival rate was significantly higher than that in control group: 71.0% (44/62) vs. 51.9% (28/54), and there were statistical differences (P<0.05). Further analysis of patients with recurrent 3 years after operation, the incidence of local recurrence and extensive peritoneal metastasis in observation group was significantly lower than that in control group: 40.4% (19/47) vs. 68.9% (31/45), and there was statistical difference (P<0.01). Conclusions It is a safe and effective treatment for intraoperative fluorouracil implant combined with raltitrexed regional chemotherapy to inhibit local recurrence and peritoneal metastasis in patients with advanced gastric cancer.
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Objective To investigate the effects of Shugan Huoxue Huatan Decoction (SHHD) on non-alcoholic steatohepatitis (NASH) of rats. Methods 32 SD male rats were randomly allocated into 4 groups:normal group, model group, Dongbao Gantai (DBG) group, SHHD group. NASH model was induced by hyperlipid diet. General condition and pathological changes of liver tissues were observed. Liver function, blood lipids, liver tissue Hyp and serum LEP were dectected. Fasting insulin resistance index (FIRI) was evaluated according to the levels of fasting blood glucose (FBG) and fasting serum insulin (FINS). Results Compared with model group, SHHD had the effects of improving liver function and blood lipids, decreasing liver tissue Hyp, LEP and FIRI siginificantly. Conclusion Inhibiting leptin and insulin resistances, enhancing the lipid metabolism are probably the main mechanisms of SHHD in preventing and treating NASH.