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Objective To investigate the effects of baicalin on the apoptosis and cell migration of intestinal epithelial cells 6 (IEC-6) induced by lipopolysaccharide (LPS) . Methods LPS was adopted to establish LPS-induced cells injury model. Methyl thiazolyl tetrazolium ( MTT) assay and Elisa kits were used to evaluate the effect of LPS on the tumor necrosis factor alpha ( TNF-α) and interleukin 6 ( IL-6) concentrations of IEC-6. Hoechst 33258 staining and flow cytometry were used to evaluate the effect of baicalin ( in the dosage of 2.5, 5.0, 10.0 μg/mL ) on the cell apoptosis. Cell migration was observed and calculated on a wounding model of IEC-6 cells induced by a pipette tip. Results Compared with the control group, the survival rate of IEC-6 in 1.0 μg·mL-1 LPS group was much lowered, TNF-α and IL-6 concentrations were significantly increased, the apoptotic rate of IEC-6 was increased and the cell migration activity was significantly decreased ( P<0.05) . Compared with the model group, IEC-6 apoptosis was relieved to various degrees in baicalin groups, the apoptotic rate of IEC-6 was obviously decreased (P<0.05) and cell migration activity was increased significantly (P<0.01) in 10.0 μg·mL-1 baicalin group. Conclusion Baicalin has the capacity of protecting IEC-6 from LPS-induced injury and increasing cell migration activity.
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Objective To investigate the cytokines levels in peripheral blood of ulcerative colitis (UC) patients at active stage, and to observe the effect of baicalin at various concentrations on the cytokines. Methods Quantitative polymerase chain reaction (Q-PCR) assay was used for the detection of interleukin 4 receptor (IL4R), IL6R, IL23R and RAR-related orphan receptor C (RORC) expression in single peripheral mononuclear cell of UC patients. Enzyme-linked immunosorbent assay (ELISA) was used to examine the serum levels of interferon gamma (IFN-γ), IL-4, IL-5, IL-6, IL-10, and transforming growth factor beta (TGF-β) levels in UC patients, and was applied for in-vitro detection of these indicators after baicalin intervention. Results Q-PCR results showed that IL4R, IL6R, IL23R, RORC gene expression levels in UC patients were increased to various degrees as compared to diarrhea-predominant irritable bowel syndrome (IBS-D) patients and the healthy volunteers, and then the levels were decreased to various degrees after intervention by baicalin at different concentrations, in particular at 20 and 40μmol/L. The results of ELISA showed that serum levels of cytokines of IFN-γ, IL-5, IL-6 in UC patients were increased to various degrees while IL-4, IL-10 and TGF-β1 was decreased as compared to IBS-D patients and the healthy volunteers. Baicalin at the concentrations of 20 and 40 μmol/L had an effect on decreasing IFN-γ, IL-5, IL-6 and on increasing IL-4 and IL-10. Conclusion Higher concentrations of baicalin show obvious effect on inhibiting RORC and IL23R expression, on decreasing IFN-γ, IL-5, IL-6 levels, and on increasing IL-4, IL-10 and TGF-β1 levels, which indicated that the therapeutic mechanism of baicalin in relieving ulcerative colonic inflammation is related with the regulation of immune function.
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Objective To investigate the inhibitory effect of baicalein on mycoplasma pneumonia and its the protective mecha -nism in body , and to provide scientific experimental basis for prevention and treatment of mycoplasma pneumoniae infection .Methods The mycoplasma pneumoniae and baicalein treated BALB /c mice lung tissues were stained with hematoxylin eosin (HE), and his-topathological grading .Minimum inhibitory concentration ( MIC) of baicalein on mycoplasma pneumonia was determined by real-time quantitative polymerase chain reaction ( PCR) .The expression of P 1 adhesion molecules mRNA and protein in lung tissue of BALB /c mice was determined with reverse transcription-PCR and Western blot .The expression of epidermal growth factor ( EGF) mRNA and protein in lung tissue was detected by quantitative RT-PCR and immunofluorescence .Results Baicalein significantly reduced the my-coplasma treated mice's lung tissue pathological score .The minimal inhibitory concentration of baicalein was 32 μg/ml.Baicalein sig-nificantly downregulated P 1 gene transcription and protein translation , and upregulated EGF gene transcription and protein expression . Conclusions Baicalein shows significant resistance to mycoplasma pneumoniae , and can protect the body against mycoplasma damage by inhibiting the expression of P 1 protein and promoting the expression of EGF protein .
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Objective To explore the effects of bacailin on the angiogenic regulating factor and migration activity of laryngeal cancer cells.Methods Hep-2 was cultured in vitro,(1)in the groups adding phorbol ester,gelatin-zymogram and Transwell assay were used to observe the effects of bacailin on the gelatinase activity of matrix metalloproteinase (MMP) and the migration ability of Hep-2,respectively; (2)in the group without phorbol ester,enzyme-linked immunosorbent assay (ELISA) was used to detect the effects of bacailin on the protein level of endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF).Results Among the control group,the low-dose group,the middle-dose group,and the high-dose group,the activity of MMP-3 of each corresponding group was (100.0 ±0.0) ％,(95.0 ±4.9) ％,(93.1± 6.2)％,and (90.7 ± 7.3)％ with statistically significant difference (t =2.123,2.309,2.412,P ＜0.05) ; the activity of MMP-9 of each corresponding group was (100.0 ± 0.0) ％,(92.3 ± 7.5) ％,(89.5 ± 9.3) ％,and (85.6 ± 6.1) ％ with statistically significant difference (t =2.253,2.302,3.708,P ＜0.05 or P ＜0.01) ;the protein level of VEGF of each corresponding group was (242.7 ±9.5)ng/L,(230.6 ± 12.7)ng/L,(212.1 ± 15.9)ng/L,and (184.2 ± 23.5)ng/L with statistically significant difference (t =2.408,3.733,4.146,P ＜0.05 or P ＜0.01) ; the protein level of bFGF of each corresponding group was (190.7±8.2) ng/L,(181.2±13.0) ng/L,(169.9±17.3)ng/L,and (140.7±9.2)ng/Lwith statistically significant difference (t =2.330,2.922,5.145,P ＜0.05 or P ＜0.01) ; Cellular migration rate of each corresponding group was (38.3 ± 5.6) ％,(32.9 ± 7.1) ％,(27.9 ± 4.4) ％,and (23.3 ± 6.0) ％ with statistically significant difference (t =2.141,3.365,4.027,P ＜ 0.05 or P ＜0.01).Conclusions Bacailin could suppress the migration of Hep-2 through inhibiting expression of angiogenic regulating factor,which might has clinical value in the treatment of laryngeal cancer.
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[Objective] To observe the effect of Baicalin on the expression of Toll-like receptors (TLRs) induced by chlamydia pneumoniae (CPN). [Methods] The endothelial cells of umbilical veins (ECV-304) was cultured in-vitro in 6-well culture plate (3 double-wells as a group). When a single layer formed, CPN or CPN + baicalin (0.48 g/L or 0.24 g/L) was added, serving as the model group and baicalin groups respectively. The wells without adding agents served as the normal group. After 5 days of culture, the intracellular growth of CPN in ECV-304 was examined by monoclonal antibody fluorescence labeling method and Giemsa stain, and the expression of TLR2 mRNA and TLR4 mRNA was detected by reverse transcriptase polymerase chain reaction (RT-PCR) in the model group. Protein expression of TLR2 on ECV-304 cell surface in the cultured supernatants was detected with flow cytometer. [Results] The intracellular growth of CPN can be found in ECV-304. The expression of TLR4 mRNA was undetectable, and high expression of TLR2 mRNA and TLR2 protein existed in ECV-304. Baicalin in high doses could lower the expression of TLR2 protein. [Conclusion] Baicalin has an inhibitory effect on the high expression of TLR2 protein in ECV-304 stimulated by CPN.