| Summary: | 碩士 === 大仁科技大學 === 生物科技研究所 === 98 === In this study, two types of red mold rice extract were analyzed for their effect on human umbilical vein endothelial cell proliferation and cytomorphology, and effect on thrombomodulin (TM) expression in tumor necrosis factor treatment (TNF-α) of HUVEC. First, we used Monascus purpureus TJ 520 to ferment Indica rice, and then extract the yielded red mold rice by ethanol and ethyl acetate. We used HPLC to analyze the concentration of monacolin K. The concentration of monacolin K in ethanol extract and ethyl acetate extract was found to be 37.4 mg/g and 13.9 mg/g, respectively. In addition, we used the MTT method to determine the viability of red mold rice extract treated HUVEC. Both extracts, at 1 μg/mL, did not affect the growth of HUVEC. The red mold rice extract treatment was found to protect the HUVEC cells against TNF-α (10 ng/mL) stimulation by exhibiting no significant change in cell morphology. In comparison, the untreated HUVEC cells revealed characteristic expansion of their cell cytoplasm at a result of TNF-α-induced changes in cell morphology. Red mold rice extract was also found to inhibit the translocation of p65 subunit of NF-κB from the cytosol to the nucleus. Using Enzyme-linked Immuno- sorbent Assay, the red mold rice extract was determined to suppress TNF-α-induced inhibition of TM expreession. Furthermore, both ethanol and ethyl acetate extract of red mold rice were also fround to inhibit TNF-α-induced activation of JNK and ICAM-1. In addition, red mold rice extract can inhibit TNF-α-induced formation of inflammation-associated material NO. Western blotting observation, 6 hours after treatment of HUVEC cell with TNF-α, and with or without red mold rice extract, indicated that the combined treatment efficiently supress the expression of NF-κB p50, κB p65 and Rac-1. We proposed that the red mold rice extract could suppress the effect of TNF-α on the TM expreession of HUVEC, at the same time, inhibited the expression of Rac-1,which in turns activated the NF-κB and affected the subsequent modulation of protein expression, and finally, resulted in mitigation of the TNF-α induced inflammation.
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