Speaker: Wei-Zhi Hsu Advisor : Jun-Yi Yang 2017/05/02 Ganoderma lucidum reduces obesity in mice by modulating the composition of the gut microbiota Speaker: Wei-Zhi Hsu Advisor : Jun-Yi Yang 2017/05/02

low-grade inflammation Obesity Insulin resistance type 2 diabetes… Obesity-associated low-grade inflammation Composition of the gut microbiota are associated with the development of obesity and its associated metabolic disorders. (Science 341, 1241214 (2013)) Obesity-induced gut dysbiosis caused by either environmental or genetic factors impairs intestinal integrity.(Gut 58, 1091–1103 (2009))

Gut microbiota The endotoxin lipopolysaccharide (LPS) from intestinal Gram-negative bacteria get into the bloodstream, leading to metabolic inflammation and insulin resistance (Invest. 115, 1111–1119 (2005).)

Obesity Pro-inflammation LPS : Lipopolysaccharide LBP : Lipobinding protein Journal of Molecular Endocrinology 51:2 ,R51–R64, 2013

Obesity Pro-inflammation, insulin resistance November 2015 | Volume 6 | Article 341

Previous study Triterpenes and polysaccharides isolated from G. lucidum inhibit adipocyte differentiation and produce hypoglycaemia effects in diabetic mice. Proteoglycans isolated from G. lucidum fruiting bodies induce antidiabetic, antihyperlipidemic and antioxidant activities.

Pro-inflammation cytokines Aim How G. lucidum produces any effect on body weight and obesity-related disorders? Obesity Insulin resistance Imflammation Intestinal integrity Pro-inflammation cytokines Gut microbiota Endotoximia (LPS) TLR4 Insulin resistance

Pro-inflammation cytokines Obesity Insulin resistance Imflammation Intestinal integrity Pro-inflammation cytokines Gut microbiota Endotoximia (LPS) TLR4

Water extract of G. lucidum mycelium (WEGL) 400 g of dried G. lucidum mycelium / 10 L water agitation at 150 r.p.m. for 30 min at 121 °C cooled to room temperature (22 °C) centrifugation at 5,894g for 30 min at 4 °C The supernatant was concentrated at 65 °C to obtain a final volume of 2 L.

WEGL prevents HFD-induced obesity in mice mice fed for 2 months chow diet HFD 100 ml water or WEGL at 2, 4 or 8% (Everyday) standard chow diet : 13.5% of energy from fat high-fat diet ( HFD ) : 60% of energy from fat

WEGL prevents HFD-induced obesity in mice

Adipose size and liver lipid content oil red O staining : lipid Dye WEGL prevent HFD-induce obesity and decrease adipose size and liver lipid content

Pro-inflammation cytokines Obesity Insulin resistance Imflammation Intestinal integrity Pro-inflammation cytokines Gut microbiota Endotoximia (LPS) TLR4

Detect inflammation gene in HFD-fed mice with WEGL Pro-inflammatory cytokines Anti-inflammatory cytokines TNF-α, IL-1β, IL-6, PAI-1 IL-10 Measure messenger RNA of hepatic and adipose

Pro-inflammatory cytokines

Anti-inflammatory cytokines WEGL reduces inflammation gene expression in HFD-fed mice

Pro-inflammation cytokines Obesity Insulin resistance Imflammation Intestinal integrity Pro-inflammation cytokines Gut microbiota Endotoximia (LPS) TLR4

WEGL reduce serum endotoximia

Pro-inflammation cytokines TLR4 JNK IKK-β IKB- α phopholation NFk-β Pro-inflammation cytokines

WEGL reduces endotoxemia and insulin resistance in HFD mice

Pro-inflammation cytokines Obesity Insulin resistance Imflammation Intestinal integrity Pro-inflammation cytokines Gut microbiota Endotoximia (LPS) TLR4

Classification of mice with WEGL or not

WEGL alters gut microbiota composition in HFD-fed mice

WEGL faecal transplants reduce obesity transferred the microbiota of WEGL-treated mice to HFD-fed mice Chow->HFD HFD->HFD Chow+8% WEGL ->HFD HFD+8% WEGL ->HFD

Compare with different organ weight Obesity and fat accumulation are reversed by faecal transplantation from WEGL-treated mice to HFD-fed mice

Analysis pro-inflammatory cytokines following faecal transplantation from WEGL-treated mice

Gut microbiota influence intestinal integrity in HFD mice Gut microbiota decrease tight-junction protein in HFD mice

Analysis of gut microbiota following faecal transplantation

Analysis of gut microbiota following faecal transplantation

Polysaccharides analysed from a WEGL

Effect of WEGL polysaccharide fractions on body weight and fat accumulation in HFD-fed mice

Summary Ganoderma lucidum can reduce obesity. Polysaccharide of WEGL play a important role of reduce obesity.