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Objective:The effects of anemoside B4 on endometritis rats were studied through in vivo and in vitro experiments. Method:Animal experiments used 25% phenol glue to prepare endometritis models. 60 female SD rats were randomly divided into blank group， model group， Kushen gel group（0.005 g·kg-1），anemoside B4 gel low，medium and high dose groups（0.005，0.01，0.02 g·kg-1），10 rats in each group，except for the blank group，rats in each group were injected with 25% phenol glue into their vagina every 2 days，and the modeling continued for 30 days. Administration started on the day after modeling. Anemoside B4 gel low， medium and high dose groups were administered rectal daily，Kushen gel group was given daily vaginal administration. The blank group and model group were given the same amount of normal saline in the same way for 30 consecutive days. After the last administration，the uterus and its attachments of each group of rats were taken to analyze the uterine morphology and index. Hematoxylin-eosin （HE） staining was used to observe the pathological changes of rat uterus. Real-time PCR was used to detect tumor necrosis factor-α （TNF-α），interleukin-1β （IL-1β），and interleukin-6 （IL-6），signal transduction protein 130 （gp130），signal transduction and transcription activator 3 （STAT3）mRNA expression. Detection of IL-6 and STAT3 protein expression in rat uterus by Western blot. In cell experiments，lipopolysaccharide （LPS）was used to induce rat endometrial epithelial cells to prepare an in vitro inflammation model， and Real-time PCR was used to detect the expression of IL-6，gp130 and STAT3 mRNA in each group of rat endometrial epithelial cells. Result:The results of animal experiments showed that compared with the blank group， the model group had inadequate uterine cavity adhesions， endometrial edema and hyperemia. Compared with model group， there was no adhesion in the uterine cavity of the rats in the high dose anemoside B4 gel group and the Kushen gel group. The uterine tissue was relatively complete， and the uterine pathological structure was significantly improved. Compared with the blank group， the uterine index of the model group was significantly increased（P<0.05）， the expression of IL-1β mRNA in the uterine tissue was significantly increased （P<0.05）， the expression of mRNA and protein of IL-6 and STAT3 in the uterine tissue significantly increased （P<0.05）. Compared with model group， the uterine index in anemoside B4 gel high dose group was significantly reduced （P<0.05）， and the mRNA and protein expression levels of IL-6 and STAT3 in the uterine tissue were significantly reduced （P<0.05）. There was no statistically significant difference between the TNF-α and IL-1β mRNA expression compared with the model group. Cell experiment results showed that compared with the blank group， the mRNA expression of IL-6 and gp130 in model group endometrial epithelial cells was significantly increased （P<0.01）， STAT3 mRNA expression was significantly increased （P<0.05）. Compared with model group， the mRNA expression levels of IL-6， gp130 and STAT3 in anemoside B4 high dose group decreased significantly （P<0.05）. Conclusion:Anemoside B4 can improve the inflammatory response of chronic endometritis in rats and reduce the release of inflammatory factor IL-6. The mechanism may be related to the down-regulation of IL-6/STAT3 pathway.
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Objective:To explore the efficacy and mechanism of Qingfei Huatan Tang on chronic obstructive pulmonary disease (COPD). Method:The rat model of COPD was established through smoke inhalation combined with lipopolysaccharide (LPS) and pulmonary compound injection. After successful modeling, the rats were randomly divided into 6 groups, namely the control group, the COPD model group, low, medium and high-dose Qingfei Huatan Tang groups and the ambroxol group. After 28 days of modeling, the drug was administered. Low, medium and high-dose Qingfei Huatan Tang (7.5, 15, 30 g·kg-1) and ambroxol (35 mg·kg-1) were administered continuously for 14 days. Immunohistochemistry and Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) were used to detect protein expression and mRNA expression of cystic fibrosis transmembrane conductance regulator (CFTR) in pulmonary fibrosis. NCI-H292 cells were induced by LPS to establish a mucus hypersecretion model. The experiment was divided into 8 groups, namely the blank control group, LPS group, LPS+10% fetal bovine serum group, LPS+ physiological serum group, LPS+5% drug serum group, LPS+10% drug serum group, LPS+20% drug serum group and LPS+AG490 group. Immunofluorescence, Western blot and Real-time PCR were used to observe the protein and mRNA expressions of CFTR in NCI-H292 cells after LPS stimulation, and western blot was used to detect the expression of tyrosine kinase 2/transcription factor 3 (JAK2/STAT3) signaling pathway in NCI-H292 cells after LPS stimulation. Result:There were a large number of brown particles around the lumen of lung tissues in the normal group, with increased COPD expression. There were a few brown particles around the lumen of lung tissues in the model group compared with the normal group, with decreased COPD expression. Compared with the normal group, mRNA and protein expressions of CFTR in the lung tissues of the COPD model group were significantly decreased (P<0.05). Compared with the model group, mRNA and protein expressions of CFTR in the lung tissues of low, medium and high-dose Qingfei Huatan Tang groups (P<0.05). Compared with the blank control group, mRNA and protein expressions of CFTR in NCI-H292 cells of the LPS group (P<0.05), with significant increases in protein expressions of p-JAK2 and p-STAT3 (P<0.05). Compared with the model group, 5%, 10%, 20% Qingfei Huatan Tang-containing serum groups showed significant increases in mRNA and protein expressions of CFTR, but with significant decreases in p-JAK2, p-STAT3 protein expressions (P<0.05, P<0.01). Conclusion:Qingfei Huatan Tang up-regulated CFTR in the treatment of COPD by inhibiting JAK2/STAT3 pathway.
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Objective@#To explore the inhibitory effect and possible mechanism of Baicalin on the human tongue squamous cell carcinoma cell line SCC15 and to provide a new idea and experimental basis for the clinical prevention and treatment of tongue squamous cell carcinoma.@*Methods @#SCC15 cells cultured in DMEM alone were used as the control group, and SCC15 cells cultured in 20 mg/mL baicalin solution were used as the baicalin group. Scratch tests and Transwell migration tests were performed to detect changes in cell migration ability, and flow cytometry was used to detect changes in the cell cycle. Western blotting was used to detect differences in the phosphorylation levels of signal transduction and transcription activator 3 (STAT3).@*Results @# Compared with the control group, the scratch test and the Transwell migration test showed that the cell migration ability of cells in the baicalin group was significantly decreased (t=4.927, P=0.008); flow cytometry showed that the number of cells of the baicalin group increased in the G0/G1 phase (t=9.893, P=0.001), decreased in the S phase (t=8.528, P=0.001), and decreased in the G2/M phase (t=3.550, P=0.024); Western blotting results showed that the STAT3 protein of SCC15 cells in the baicalin group decreased (t=3.550, P=0.024), and the phosphorylation level significantly decreased (t=8.262, P=0.001).@*Conclusion @#Baicalin inhibits the human tongue squamous cell carcinoma cell line SCC15, and its mechanism may be related to a decrease in STAT3 pathway phosphorylation activity.