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Effect of EGCG on UBC9 expression in inflammatory model of A549 cells |
CAO Liuxia, WU Jiansong, XU Xiaodan, and HE Dan |
Department of Infectious Diseases, General Hospital of the PLA Rocket Force, Beijing 100088, China |
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Abstract Objective To observe the effect of EGCG on the expression of UBC9 in the A549 inflammatory model, and investigate its correlation with the anti-inflammatory mechanism of EGCG.Methods A549 cells were randomly divided in vitro into four group: control group, EGCG group, LPS group, and EGCG plus LPS group. The proliferative activity of A549 cells, which were incubated with different concentrations of EGCG(50~400 μg/ml) and LPS(10 ~50 μg/ml), was detected by MTT method to select the optimum concentration for experiment. The relative expressions of UBC9 protein and mRNA in each group were determined with immunohistochemical staining, Western blot and real-time-PCR, and compared.Results MTT showed that 200 μg/ml EGCG had the strongest inhibitory effect on proliferation of A549 cells(F=16181.18, P<0.001), while 25 μg/ml LPS had the strongest promoting effect (F=237.38, P<0.001). As observed with immunohistochemistry and Western blot, EGCG down-regulated the expression of UBC9 protein(P<0.001) while LPS up-regulated the expression(P<0.001) compared to control group. Compared to LPS group, the expression of UBC9 protein in EGCG plus LPS group was reduced significantly(P<0.001). Similarly, Real-time-PCR revealed that the expression of UBC9 mRNA decreased in EGCG group(F=89.34, P<0.001), but increased in LPS group(F=225.00, P<0.001) compared with control group. Compared to LPS group, the expression of UBC9 mRNA in EGCG plus LPS group was inhibited and reduced significantly(F=625.00, P<0.001).Conclusions EGCG can down-regulate the expression of UBC9 in the inflammatory model induced by LPS.The anti-inflammatory mechanism of EGCG might be mediated by suppression of UBC9 protein and mRNA expression.
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Received: 05 September 2016
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