Objective

To establish an ischemia-reperfusion injury model of rat cerebral microvascular endothelial cells (MVECs) in vitro, and to explore the relationship between nuclear factor-kappa B (NF-κB) and the protective effects of Qingkailing effective components (hyocholic acid, taurocholic acid, baicalin, jasminoidin, Pinctada martensii) on MVECs.
Methods

Brain MVECs of male rats were digested with trypsin and subcultured, then the content of MVECs was adjusted to 1×105/mL and the MVECs were divided into normal control group, untreated group, hyocholic acid group, taurocholic acid group, baicalin group, jasminoidin group, Pinctada martensii group and nimodipine group, with six holes in each group. Except for the normal control group, the MVECs in the other groups were exposed in oxygen and glucose deprivation (OGD) circumstance in vitro to simulate ischemia-reperfusion injury. Immunocytochemical staining and image analysis system were used to observe the expression of NF-κB protein.
Results

Under a light microscope, the nuclei of MVECs in the normal control group were blank. Staining intensity of NF-κB protein in the nucleus in the untreated group was much deeper than that in the endochylema, with NF-κB shifted to nucleus after activation; a small quantity of NF-κB protein were expressed in the border of nucleus next to endochylema in groups of Qingkailing effective components, and the NF-κB protein expression was weaker than that in the untreated group. With the image analysis, we found that transmittance of nucleus and endochylema in the untreated group was significantly lower than that in the normal control group (P<0.01). Transmittance of nucleus and endochylema in the treated groups was higher than that in the untreated group (P<0.05, P<0.01).
Conclusion

Qingkailing effective components have significant effect in inhibiting NF-κB protein transferring from endochylema to nucleus in vitro.