| Summary: | 碩士 === 國立中興大學 === 食品暨應用生物科技學系所 === 102 === Recently, Pleurotus eryngii and Hypsizigus marmoreus are important edible mushrooms in Taiwan. Grifola frondosa is a popular material for study and contains various bioactive substances and nutrients. Antrodia salmonea is a new species of genus Antrodia as a basidiomycete first identified in 2004 in Taiwan. Recent research has pointed out that, new components isolated from A. salmonea have good effect on anti-inflammatory and antitumor. Nonalcoholic fatty liver disease (NAFLD) is one of the most common liver diseases currently in worldwide, including fatty liver, nonalcoholic steatohepatitis (NASH), fibrosis, necrosis and cancer. NAFLD is caused by fat metabolic imbalance in liver resulting from excessive fat (triglyceride, TG) accumulation in hepatocytes. In this study, we investigated the effect of water (W) and ethyl acetate (EA) fractions of 70 % ethanolic extracts from P.eryngii (PE), H.marmoreus (HM), G.frondosa (GF) fruiting bodies and A.salmonea (AS) mycelia on anti-inflammatory properties and potential for improving on NASH.
The results showed that the content of total phenols and flavoniods in PEW, PEEA, HMW, HMEA GFW, GFWA, ASW and ASEA were 0.31-8.10 GAE mg/g and 0.22-10.95 RE mg/g, respectively. According to the results of MTT assay, the effect of different extracts on cell viability of RAW264.7 cells showed dose- and time-dependent suppression. The anti-inflammatory activity evaluation, we investigated the effect of extracts on LPS-induced inflammatory mediators production in RAW264.7 cells. The superior effects on reducing NO, TNF-α, IL-1β and IL-6 productions caused by different extracts which were 750 μg/mL, 100 μg/mL PEEA, 750 μg/mL ASW and 750 μg/mL GFEA, respectively. Except HMEA, most of extracts could increase the IL-10 production on LPS-induced activated. RAW264.7 cells. These results indicated that 70 % ethanolic extracts from PE, HM, GF and AS had anti-inflammatory properties in LPS-induced RAW264.7 cells.
The cell model of free fatty acids (FFAs) mixture (oleate : palmitate= 2 : 1) induced HepG2 cells was further established to mimic NASH. There were two groups : steatotic/simultaneous group (prophylaxis group) was incubated with extracts and FFAs mixture at same time for 24 h and steatotic/non-simultaneous group (therapy group) that was treated with FFAs mixture for 24 h prior to 24 h incubation with extracts. According to the results of Oil red O stain, GFW, GFEA, ASW and ASEA may decrease the TG contents on FFAs mixture-induced HepG2 cells. In addition, all of four extracts all significantly decreased ROS production in HepG2 cells caused by FFAs mixture. Besides, GF and AS extracts were able to release glycerol from the fat over-loadding HepG2. These results demonstrated that extracts showed better effect on prophylaxis group than therapy group. According to the results of western blotting, GF and AS down-regulated acyl-CoA carboxylase (ACC), fatty acid synthase (FAS) and SREBP-1c (sterol regulatory element binding protein 1c) expression and enhanced PPARα (peroxisome proliferator activated receptor α) protein expression. These results suggested that GFW, GFEA, ASW and ASEA were effective in protecting liver from fat accumulation which may lead to NASH.
In conclusion, the 70 % ethanolic extracts from PE, HM, GF fruiting bodies and AS mycelia showed strong anti-inflammatory property. GF and AS can decrease TG accumulation in HepG2 cells. It seems that GF and AS might have potential for improvement of NASH.
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