ABSTRACT
Objective To evaluate the protective effect of curcumin against reperfusion injury of mouse renaltubular epithelial cells in vitro and the related mechanisms. Methods The renal tubular epithelial cells(TECs)were isolated by an improved Percoll density gradient centrifugation method. The cultured TECs were divided into blank group, hypoxia reperfusion group, low-concentration curcumin (5μmol/L) and high? Concentration curcumin (25μmol/L) groups. Apoptoticrate of TECs was examined with FACS, and them RNA and protein levels of TLR4 were detected with RT-PCR and Westernblotting analysis, respectively. Meanwhile, expression of IL-6, IL-1ß, TNF-α and MCP-1 mRNA in TECs was examined by RT-PCR, and their contents in the supernatants of TECs were measured by ELISA. Results The purity of the isolated TECs washigher than 95%, and the cell viability was higher than 93%. Compared with hypoxia reperfusion group, the two curcumin groups exhibited significantly lower apoptotic rates (P<0.01) and lower levels of TLR 4m RNA and protein (P<0.01), inhibited P65 phosphorylation, and significantly decreased levels of IL-6, IL-1ß, TNF-α and MCP-1 in the TECs (at both mRNA and protein levels) and their supernatant (P<0.05orP<0.01). Conclusion Curcumin can protect TECs from reperfusion injury in vitro, which might be associated with the inhibition of TLR4 expression and the consequent attenuation of TLR4- triggered inflammatory damages.