益生菌 Lactobacillus casei Shirota 與高油高糖飲食對 Vhlh 基因缺失小鼠脂質代謝與免疫反應之影響

Effects of probiotic Lactobacillus casei Shirota and high-fat-sucrose diet on lipid metabolism and immune responses in Vhlh gene defect mice

葉雨晴(Yu-Qing Ye)

指導教授 ： 林璧鳳

國立臺灣大學/生命科學院/生化科技學系/碩士(2020年)

https://doi.org/10.6342/NTU202004212

摘要

台灣末期腎臟病疾病盛行率為全球第一，已知過重及肥胖所造成的代謝異常如高血糖、高血脂會增加腎病變的風險。因此，本研究欲探討代田菌 (Lactobacillus casei Shirota, LcS) 的補充對高油高糖致肥胖的Vhlh基因缺失小鼠脂質代謝的影響。實驗一先以自發性腎炎致腎癌之Vhlhdel/del小鼠，將離乳後5週齡開始以管餵方式補充LcS，觀察生命期，結果發現LcS可以顯著延長Vhlhdel/del小鼠生命期。實驗二以高油高糖飲食致肥胖Vhlh+/del小鼠，探討補充LcS對醣類與脂質代謝及免疫調節的影響。結果顯示，餵飼LcS8週後能顯著降低小鼠體重，延緩血糖的上升並減緩胰島素阻抗。肝臟三酸甘油酯及膽固醇在高油高糖飲食下顯著增加，補充LcS可顯著降低肝臟總膽固醇含量。補充LcS對盲腸及糞便的指標菌門的結果，高油高糖飲食顯著降低盲腸的厚壁菌門及擬桿菌門，補充LcS有降低厚壁菌門並提升擬桿菌門的趨勢；糞便中擬桿菌門相對量達顯著差異，而有降低厚壁菌門與擬桿菌門比例的趨勢。指標菌屬評估，高油高糖飲食糞便中乳酸菌屬有降低趨勢，LcS補充增加盲腸及糞便中乳酸菌屬量，WT小鼠糞便達顯著差異；高油高糖飲食降低盲腸及糞便中比菲德氏菌屬，LcS補充可回升比菲德氏菌屬數量，也在糞便達顯著差異。腸道免疫細胞群結果顯示PP中，LcS補充可顯著增加脾臟中CD4+ T細胞比例。細胞激素方面，高油高糖飲食促進MLN及脾臟細胞IL-6分泌以及PP的TNF-α的分泌。補充LcS降低脾臟IL-6的分泌。高油高糖飲食顯著降低IL-10的分泌，補充LcS有增加Vhlh+/del小鼠調節型細胞激素IL-10生成的趨勢，同時降低IL-4的產生。綜上所述，補充LcS對於高油高糖飲食致肥胖之Vhlh+/del小鼠，能有效改善其血糖及脂質代謝，並調節體內免疫反應，可能有助延緩慢性腎炎致腎癌Vhlhdel/del小鼠生命期。

關鍵字

代田菌 ； 黏膜免疫系統 ； 肥胖 ； 腸道菌群

並列摘要

The incidence of kidney cancer is increasing by approximately 2% per year. The prevalence of end-stage renal disease in Taiwan ranks the first place in the world. This increasing incidence is thought to be partly due to an increased prevalence of obesity, which may cause metabolic abnormalities like hyperglycemia and dyslipidaemia, the risk factors of nephropathy. This study aimed to investigate the effects of Lactobacillus casei strain Shirota (LcS) on lipid metabolism and rephropathy. The experiment 1 demonstrated that oral administration of LcS improved the life span of Vhlhdel/del mice. To investigate whether LcS may favourably affect metabolic abnormalities in obese Vhlh+/del mice. Herein, high-fat-sucrose diet (HFS)-induced obese mice were supplemented with oral administration of LcS for 48 weeks. The result showed that the body weight of Vhlh+/del mice significantly decreased, fasting serum glucose level and insulin resistance were improved compared to the HFS group. Furthermore, the gut microbiota in fecal and cecum contents from mice fed high-fat-sucrose diet exhibit significantly reduced the population of Firmicutes and Bacteroidetes compared to the that of control diet group. Probiotic LcS increased Bacteroidetes, decreased Firmicutes in both on fecal and cecum. Lactobacillus and Bifidobacterium significantly decreased in the HFS diet group. LcS supplement elevated the population of Lactobacillus and Bifidobacterium in fecal significantly, and also increased Lactobacillus in cecum in Vhlh+/del mice. In addition, after long-term induced by HFS diet which in turn triggers chronic low-grade inflammation, results of immune cells population and secreting cytokines showed that the proinflammatory cytokine IL-6 increased in mesenteric lymph nodes (MLN) and spleen in the HFS group. LcS group significantly decreased on IL-6 in spleen compared to the HFS group. The HFS diet also decreased regulatory cytokine IL-10 in MLN and spleen, which increased with LcS supplement. And declined IL-4 effectively after LcS supplement in spleen.
In conclusion, LcS as a probiotic supplement, greatly improve the carbohydrate and lipid metabolism, gut microbiota, immune responses affected by the HFS diet. And LcS supplement also may improved the life span of Vhlhdel/del mice.

並列關鍵字

Lactobacillus casei strain Shirota ； obesity ； mucosal immune system ； gut microbiota

參考文獻

辜祥霖 (2016)，不同葉酸含量對高油飲食小鼠腎臟的影響，國立臺灣大學生化科技學系 碩士論文

Amabebe, E., F. O. Robert, T. Agbalalah, and E. S. F. Orubu. 2020. 'Microbial dysbiosis-induced obesity: role of gut microbiota in homoeostasis of energy metabolism', Br J Nutr, 123: 1127-37.

Amar, J., C. Chabo, A. Waget, P. Klopp, C. Vachoux, L. G. Bermúdez-Humarán, N. Smirnova, et al. 2011. 'Intestinal mucosal adherence and translocation of commensal bacteria at the early onset of type 2 diabetes: molecular mechanisms and probiotic treatment', EMBO Mol Med, 3: 559-72.

Anders, H. J., K. Andersen, and B. Stecher. 2013. 'The intestinal microbiota, a leaky gut, and abnormal immunity in kidney disease', Kidney Int, 83: 1010-6.

Aronsson, L., Y. Huang, P. Parini, M. Korach-André, J. Håkansson, JÅ Gustafsson, S. Pettersson, et al. 2010. 'Decreased fat storage by Lactobacillus paracasei is associated with increased levels of angiopoietin-like 4 protein (ANGPTL4)', PLoS One, 5.