ABSTRACT
AIM: To study the roles of Toll-like receptor 4 ( TLR4 ) and TLR4 activator lipopolysaccharide ( LPS) in colonic inflammation recovery .METHODS:Normal intestinal epithelial cells were cultured with LPS in vitro. The subgroups of the intestinal epithelial cells with differential expression of TLR 4 ( low, normal and high ) were construc-ted by the technique of lentivirus transfection .The cells with normal and high expression of TLR 4 were induced by LPS for 0 h, 2 h and 4 h.Inflammatory cytokines TNF-α, IL-6 and IL-8 in the culture supernatant were detected by ELISA .The mRNA levels of TNF-α, IL-6, IL-8, IL-10 and IL-1βwere detected by qPCR .The cell mobility was also monitored by wound healing assay .RESULTS:The protein expression of TLR 4 was significantly higher after LPS treatment than that in control groups of both cells with TLR4 normal and high expression (P<0.05).The inflammatory cytokines TNF-α, IL-6, IL-8 and IL-1βat mRNA and protein levels were also significantly increased after LPS treatment compared with control group (P<0.05).The protein levels of TNF-α, IL-6 and IL-8 between the 2 groups were also different with statistical sig-nificance ( P<0.05 ) .Higher mobility was observed in the cells with TLR 4 high expression compared to control cells . CONCLUSION:LPS induction might play a role in the activation of TLR 4-mediated inflammatory pathways by up-regula-ting the expression of inflammatory cytokines at both transcriptional and translational levels .
ABSTRACT
Objective To investigate the micro-vessel density and vascular endothelial growth factor receptor 2 (VEGFR2) expression in mucosa and submucosa tissue of the patients with hemorrhoids. Methods Sixty hemorrhoids patients were randomly recruited and involved into experiment group and 20 cases of normal patients involved in control group. RASP, MASP, RSTV, RMTV, Expression of VEGFR2 and microvessel density in two groups were detected by immunohistochemistry. The expression abundance of VEGFR2 mRNA was detected by RT-PCR. Results Excessive angiogenesis was found in hemorrhoids tissue (5.59 ± 0.98) in experiment group as compared with that in control group (3.92 ± 0.81), the difference was statistically significant (P < 0.05). Staining score of hemorrhoid tissue and the control tissue were 5.95 ± 1.01, 2.32 ± 0.72, respectively (P < 0.05). Abundance of VEGFR2 mRNA expression in hemorrhoid tissue was significantly higher than that in control tissue (P < 0.05). Conclusion VEGFR2 plays an important role in angiogenesis during the development of hemorrhoids, which may be closely related to the occurrence of hemorrhoid disease and clinical symptoms.
ABSTRACT
Objective To investigate the effects of gambogic acid on proliferation,invasion and protein expression of MMP-2 in LoVo human colon cancer cell line.Methods The CCK-8 assay was used to determine the anti-proliferation ability.Transwell chamber assay was used to study the invasion of LoVo colon cancer cells.Western Blot was applied to examine the protein expression of MMP-2.Results CCK-8 assay showed that after cells treated with 1,2,4 μmol/L GA for 24 h,48 h the inhibition rates were(0.16±0.11)％,(0.24±0.08)％,(0.58±0.01)％,(0.67±0.03)％,(0.79±0.01)％ and (0.27±0.05)％,(0.69±0.09)％,(0.85±0.01)％,(0.87±0.01)％,(0.89±0.01)％,and there had statistically significant differences between control group (0 μmo/L) with experimental group (P＜0.05).Gambogic acid can effectively inhibit LoVo cells invasion at a low-dose concentration,which the numbers of cells that digested the matrigel in control group (0 μmo/L)was 120.50± 8.69 and 69.83 ±4.75 in experimental group (1 μnol/L),P＜ 0.05).Western Blot revealed that Gambogic acid can down-regulate the expression of MMP-2,which the levels of down-regulating the expression of MMP-2 after treated with 1,2,4 umol/L GA were 48.67％,74.72％and 82.70％ (P＜0.05) Conclusion Gambogic acid can effectively inhibit the growth and invasion of the LoVo cells,which may be related with down-regulating the expression of MMP-2.