寡孢根黴菌與胚芽乳桿菌共發酵天貝緩解棕櫚酸誘導人類肝癌細胞HepG2及FL83B細胞脂毒性之研究

Alleviation of Rhizopus oligosporus and Lactobacillus plantarum co-fermented tempeh on palmitic acid-induced lipotoxicity in HepG2 and FL83B cells

郭怡萍(Kuo Yi-Ping)

指導教授 ： 吳明昌

國立屏東科技大學/農學院/食品科學系所/碩士(2019年)

https://doi.org/10.6346/NPUST201900252

摘要

天貝是一種黃豆發酵產品，為印尼傳統發酵食物，主要是因為其具有多項有益的成份及抗氧化能力，天貝發酵之菌株主要為寡孢根黴菌，其屬於毛黴科家族之真菌。有研究發現，寡孢根黴菌與胚芽乳桿菌共同培養的天貝比未發酵大豆與寡孢根黴菌培養的天貝具有較高的營養和生物活性。天貝具有調節血糖和增強免疫力的活性，但糖尿病併發症和非酒精性脂肪肝的功效仍未得到闡明。因此，在本研究中，我們評估寡孢根黴菌與胚芽乳桿菌共同培養的天貝萃取異黃酮對棕櫚酸誘導的肝細胞脂質堆積和發炎反應以及脂肪生成之影響。在HepG2和FL83B細胞中使用棕櫚酸誘導脂質毒性24小時。接著使用未發酵的大豆、寡孢根黴菌培養的天貝和寡孢根黴菌與胚芽乳桿菌共同培養的天貝萃取其異黃酮，以不同的劑量來處理細胞24和48小時，以MTT試驗來測量細胞毒性。油紅染色用於測定細胞攝取的游離脂肪酸。此外，西方墨點法用於測定脂質代謝和脂肪生成以及發炎反應相關的各種標誌物的蛋白質表現。結果顯示來自寡孢根黴菌與胚芽乳桿菌共同培養的天貝萃取物異黃酮具有降低HepG2細胞存活率。此外，在所有的異黃酮萃取組別中皆能抑制棕櫚酸所誘導的HepG2和FL83B細胞內脂質堆積，另外，亦發現異黃酮萃取物能使棕櫚酸對TNF-α、NF-κB、COX-2和FASN表現具有影響。這些結果說明了從寡孢根黴菌與胚芽乳桿菌共同培養的天貝萃取的異黃酮能減低棕櫚酸誘導的HepG2和FL83B細胞中的脂毒性和脂肪生成。本研究也顯示，從天貝萃取的異黃酮具有治療非酒精性脂肪肝的潛力，值得做更進一步的研究。

關鍵字

天貝 ； 非酒精性脂肪肝 ； 棕櫚酸

並列摘要

Tempeh, which is a fermented soybean product consumed widely in Indonesia has several beneficial characteristics and antioxidative activity. Rhizopus oligosporus, a fungus of Mucoraceae family is widely used as the starter of homemade tempeh. It has been shown that R. oligosporus and L. plantarum co-inoculated tempeh contained higher isoflavones and possessed high nutritional and bioactive properties than R. oligosporus inoculated tempeh and unfermented soybean. Tempeh has the activity of regulating blood sugar and boosting immunity, but the efficacy of diabetic complications and non-alcoholic fatty liver is still not elucidated. Therefore, in the present study, we attempt to evaluate the effects of isoflavones extract from tempeh co-inoculated with Rhizopus oligosporus and L. plantarum on palmitic-induced lipid accumulation, inflammation and lipogenesis in hepatocytes. HepG2 and FL83B cells were treated with palmitate for 24 hours to induce lipid toxicity. The cells were then treated with different doses of isoflavones extract from unfermented soybean, isoflavones extract from R. oligosporus inoculated tempeh and isoflavones extract from R. oligosporus and L. plantarum co-inoculated tempeh for 24 or 48 hours. MTT assay was used to measure the cytotoxicity. Oil Red O staining was used to determine the cellular uptake of free fatty acid. In addition, western blotting was used to determine the expression levels of various protein related to lipid metabolism, lipogenesis and inflammations. Isoflavones extract from L. plantarum co-inoculated tempeh, caused a reduction in HepG2 cell viability. Furthermore, isoflavones extract in all groups reversed the palmitate-induced intracellular lipid accumulation in HepG2 and FL83B cells. Moreover, isoflavones extract was also found to be reverted the palmitic acid effect on the expression of TNF-α, NF-κB, COX-2, and FASN. These results indicated that isoflavones extract from tempeh co-inoculated with Rhizopus oligosporus and L. plantarum attenuates PA-induced lipotoxicity and lipogenesis in HepG2 and FL83B cells. In conclusion, this study suggested that isoflavones extract from tempeh has a therapeutic potential for fatty liver disease and warrants further investigation.

並列關鍵字

Tempeh ； non-alcoholic fatty liver disease (NAFLD) ； palmitic acid (PA)

參考文獻

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