PREVENTION OF NON-ALCOHOLIC FATTY LIVER DISEASE BY DECAFFEINATED GREEN TEA EXTRACT IN HIGH FAT-FED MICE
Open Access
- Author:
- Khoo, Weslie Yu
- Graduate Program:
- Food Science
- Degree:
- Doctor of Philosophy
- Document Type:
- Dissertation
- Date of Defense:
- March 26, 2019
- Committee Members:
- Joshua D. Lambert, Dissertation Advisor/Co-Advisor
Joshua D. Lambert, Committee Chair/Co-Chair
Ryan J. Elias, Committee Member
Kathleen L. Keller, Committee Member
Donna H. Korzick, Outside Member - Keywords:
- NAFLD
Non alcoholic fatty liver disease
Decaffeinated green tea extract
Green tea
PPARα
Lipid metabolism
Mitochondrial biogenesis
Inflammation
Toxicity - Abstract:
- Non-alcoholic fatty liver disease (NAFLD) is a spectrum of diseases characterized by abnormal lipid accumulation in the liver in the absence of excessive consumption of alcohol. NAFLD is a significant and growing public health problem, with a global prevalence of about 25%. NAFLD is strongly associated with obesity, insulin resistance and type 2 diabetes mellitus. Currently, there are no validated therapies for NAFLD although some studies have suggested that lifestyle interventions that promote weight loss alone or in combination with a pharmacological treatment may be beneficial. Green tea (Camellia sinensis, Theaceae) is a widely consumed beverage and its extract can be found in various herbal dietary supplements. Green tea-based dietary supplement sales in the U.S. exceeded USD 48 million in 2015. Green tea has been extensively studied for weight loss and weight maintenance effects, preventive effects against cancer, cardiovascular diseases, neurodegenerative diseases and improvements in psychopathological symptoms. There is a growing body of evidence from animal and human studies of the effects of green tea or its polyphenolic constituents (e.g. catechins) on markers related to NAFLD. Green tea was found to decrease body weight gain, fat mass, and improve dyslipidemia by lowering blood levels of cholesterol and triglycerides (TAG) in mice fed a high fat (HF) diet. In addition, liver weight as well as biochemical markers of NAFLD such as liver TAG and total lipid content in HF-fed mice were reduced by green tea or its catechins. The number of studies on the impact of green tea on NAFLD in human subjects is limited, but the results are promising. Green tea treatment improved blood markers of liver injury and, and in some cases, dyslipidemia. Some studies have however, failed to find an effect of green tea and its catechins on blood lipid levels. Therefore, more research is required to understand the underlying mechanisms of how green tea affects lipid changes and NAFLD in humans. Previous studies in our laboratory have shown that combination treatment with decaffeinated green tea extract (GTE) and voluntary exercise (Ex) reduced the development of obesity and insulin resistance in HF-fed mice to a greater extent than GTE- or Ex- treatment alone. These effects were related to increased expression of genes related to mitochondrial biogenesis in skeletal muscle and visceral adipose tissue “browning”. In addition, combined effect of GTE- and Ex- increased the expression of hepatic genes related to fatty acid oxidation. The overall purpose of this dissertation research is to examine the NAFLD preventive effects of GTE in two HF-fed mouse models. In the first study, we investigated the effects of GTE-, Ex- and the combination of both GTE- and Ex- on parameters related to NAFLD in HF-fed mice. We hypothesized that the combination of GTE- and Ex- would have greater NAFLD preventive effects than either GTE- or Ex- alone and that these effects are due to the inhibition of macronutrient digestion and the regulation of genes related to mitochondrial biogenesis, lipid metabolism and inflammation. Male C57BL/6J mice were randomized to a HF diet (60% energy from fat), HF supplemented with decaffeinated green tea extract (7.7g GTE/kg), HF plus access to a voluntary running wheel (Ex), or the combination (GTE and Ex) and treated for 16 weeks. We found that treatment of mice with the combination of GTE- and Ex- mitigated HF-induced NAFLD and was more effective than either treatment alone. The combination of GTE- and Ex- reduced plasma alanine aminotransferase, hepatic TAG and lipid accumulation, compared to either treatment alone. Mitigation of NAFLD was associated with increased fecal lipid and protein levels, reduced systemic inflammation, and higher hepatic expression of genes related to mitochondrial biogenesis. In the liver, GTE-, Ex-, and the combination-treatment groups also had higher hepatic expression of genes related to cholesterol synthesis and uptake. The magnitude of these effects was not different between mice receiving single treatments or the combination. No difference treatment effect on the hepatic expression of lipolysis-associated genes was observed. In the second study, we hypothesized that peroxisome proliferator-activated receptor alpha (PPARα) plays a role in regulating the beneficial effects of green tea on preventing NAFLD. PPARα is a transcription factor and plays a role in regulating gene expression related to lipid metabolism, gluconeogenesis, antioxidant response, and intestinal nutrient absorption. PPARα is a potential target for the therapeutic treatment of NAFLD. Although tea polyphenols were found to activate PPARα in the liver, skeletal tissues, there are still inconsistencies examining the relationship between green tea and PPARα in models of metabolic syndrome. For this second study, we used PPARα-deficient and wild-type mice of the same genetic background (C57BL/6N) to investigate the role of PPARα in the NAFLD-mitigating effects of green tea. Female PPARα−/− (KO) and PPARα+/+ (WT) were randomized to receive either a HF diet (60% energy from fat) or a GTE supplemented HF diet (60% Kcal, with 7.7g GTE/kg) for 12 weeks. We report that GTE supplementation led to 20% mortality in HF-fed KO mice, while no mortality occurred in WT mice. Furthermore, GTE-treated KO mice were observed to be lethargic, and some noticeable decreased responsiveness to touch. The dose of GTE used in our studies has been used in a number of previous mice model experiments without reported adverse effects. These results demonstrate for the first time that lack of PPARα may increase sensitivity to the potential toxic effects of GTE. Higher doses of GTE used in animal models have been shown to induce mortality and hepatotoxicity. Overall, GTE tended to prevent hepatic lipid accumulation in WT mice but not in KO mice. However, GTE increased systemic inflammation and decreased hepatic anti-inflammatory markers in KO mice. Taken together, our results suggest the role of PPARα in modulating the effects of GTE. As both studies used different sex, a post-hoc analysis of data derived from the studies described in chapter 2 and chapter 3 was done to develop a hypothesis regarding the role of sex as a mediator of the NAFLD preventive effects of GTE that can be tested in future animal model studies. Our post-hoc analysis demonstrates that GTE supplementation can affected weight gain, hepatic lipid accumulation and hepatic expression of genes related to cholesterol synthesis, mitochondrial biogenesis, and inflammation differently in HF-fed male and female mice. Collectively, we have demonstrated that GTE can prevent NAFLD in HF-fed mice in a PPARα -dependent manner. This preventive effect of GTE is further enhanced when combined with Ex. The preventive effects of GTE and Ex are related to treatment-related decreases in macronutrient absorption and inflammation, and increased expression of hepatic markers related to fatty acid oxidation and mitochondrial biogenesis. From both studies, GTE was found to upregulate genes related to hepatic cholesterol synthesis in a PPARα-dependent manner. Lastly, not only were the beneficial effects of GTE lost in KO mice, KO mice could be more sensitive to GTE toxicity. Taken together, this dissertation provides novel mechanisms which GTE exerts it’s NAFLD preventive effects.