| Rat intestinal microvascular endothelial cells were cultured in vitro and were detected by cellar immunity chemical method.Results:cells showed a typical cobblestone-like under the inverted microscope. Staining with non-specific rabbit IgG was performed as a control and no cells were positively stained.To investigate the inhibition of lactic acid on E. coli lipopolysaccharide (LPS)-activation of NF-κB p65 in rat intestinal mucosa microvascular endothelial cells (RIMMVECs). RIMMVECs, cultured in vitro, were pretreated with different concentrations of lactic acid and then exposed to LPS. Cells and cell culture media were then collected at different time intervals. Production of TNF-a and IL-6 was examined at the protein level by enzyme-linked immunosorbent assay. The influence of lactic acid on the LPS-activation of NF-κB was examined at mRNA and protein levels by real-time quantitative PCR and western blot analysis, respectively. TNF-a and IL-6 levels reached a peak at 9 h but were not different from those of the blank control; however, they were significantly lower compared with the positive control.The influence of lactic acid on the LPS-activation of NF-κB was examined at mRNA and protein levels by real-time quantitative PCR and western blot analysis, respectively.Real-time PCR analysis of NF-κB p65 mRNA levels revealed that after pretreatment with 7.5μl.ml-1,5.0μl.ml-1 or 2.5μl.ml-1 lactic acid, followed by stimulation with LPS, the NF-κB mRNA levels of the test groups were 1.51-, 2.62- and 3.00-fold greater than the blank control, respectively. Western blot analysis using antiβ-actin and anti- NF-κB p65 antibodies produced protein bands of 42 kDa and 65 kDa in each group, respectively. There was a significant difference in NF-κB p65 protein level between the positive control group and the test groups (P<0.01). There were no significant differences in the levels of NF-κB p65 between the test groups and the blank control group.
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