Obesity is becoming a major public health problem worldwide. Making charcoal from wood (“Sumi-yaki”) has been a traditional activity in the southern part of Nagano Prefecture for centuries, with activated charcoal having reported detoxifying effects. However, it is unclear whether activated charcoal also possesses anti-obesity properties. Additionally, since activated charcoal is usually alkaline and might be affected by gastric juice, we evaluated the effect of acidic activated charcoal on high-fat diet (HFD)-induced obesity. This study demonstrated that co-treatment of acidic activated charcoal with a HFD significantly improved obesity and insulin resistance in mice in a dose-dependent manner. Metabolomic analysis of cecal contents revealed that neutral lipids, cholesterol, and bile acids were excreted at markedly higher levels in feces with charcoal treatment. Moreover, the hepatic expressions of genes encoding cholesterol 7 alpha-hydroxylase and hydroxymethylglutaryl-CoA reductase/synthase 1 were up-regulated by activated charcoal, likely reflecting the enhanced excretions from the intestine and the enterohepatic circulation of cholesterol and bile acids. No damage or abnormalities were detected in the gastrointestinal tract, liver, pancreas, and lung. In conclusion, acidic activated charcoal may be able to attenuate HFD-induced weight gain and insulin resistance without serious adverse effects. These findings indicate a novel function of charcoal to prevent obesity, metabolic syndrome, and related diseases.
Introduction
Obesity has reached pandemic levels worldwide. Since 1980, the prevalence of obesity has doubled in more than 70 countries/regions and has continued to increase in most other countries. Obesity is a disease caused by excessive fat accumulation. It increases the risk of other diseases, such as cardiovascular disease, type 2 diabetes, hypertension, non-alcoholic fatty liver disease, chronic kidney disease, sleep apnea syndrome, and mood disorders. In recent years, obesity has become a major public health concern in South Asia. In some Asian countries, obesity and its related metabolic diseases is widespread in more than one-third of the population (1–3).
Making charcoal from wood (“Sumi-yaki”) has been a traditional activity in the southern part of Nagano Prefecture for centuries, with “Ina Akamatsu” charcoal was made into activated charcoal for deodorant and water purification. Previous studies have revealed detoxifying effects for activated charcoal. Meinita et al. reported that the material could remove 5-hydroxymethylfurfural, levulinic acid, and other toxic substances within 30 min (4). Elsewhere, recommendations given by poison control centers in Germany state that activated charcoal is suitable for primary toxin clearance in cases of moderate-to-severe poisoning; charcoal is especially suitable for poisons that remain for long periods in the stomach and poisons circulating between the intestine and liver (5). Neuvonen et al. also demonstrated the ability of activated charcoal to effectively bind bile acids (BAs) in vitro (6). The above observations corroborate the biological action of activated charcoal as an absorber of toxins and lipid derivatives.
It is well known that dietary composition, including fat and contaminants, as well as microbiota-produced BAs affect adiposity through multiple pathways. To date, however, the anti-obesity effect of activated charcoal has not been addressed. In a preliminary experiment, the anti-obesity effect of alkaline activated charcoal was not significant in high-fat diet (HFD)-treated mice. Additionally, the mice consuming alkaline activated charcoal looked irritable (unpublished data). We speculated that acidic activated charcoal may avoid interference of gastric juice and be more suitable than regular alkaline charcoal. Therefore, we originally developed acidic activated charcoal with a pH of approximately 5 and treated C57BL/6J mice with the acidic activated charcoal powder-containing HFD, in order to investigate whether the acidic charcoal exerts an anti-obesity effect.
Discussion
The current study showed that acidic activated charcoal could prevent HFD-induced obesity, insulin resistance, eWAT hypertrophy and inflammation, as well as enhance BAT whitening in a dose-dependent manner without any serious adverse effects. Metabolomic analysis of cecal contents revealed that neutral lipids, cholesterol, and BAs were markedly more excreted into the feces with charcoal supplementation. Consequently, the enterohepatic circulation of cholesterol/BA was promoted as evidenced by the up-regulated hepatic expression of de novo BA- and cholesterol-synthesizing enzymes (13). The current study proposes a novel function of acidic activated charcoal to prevent obesity, overnutrition, metabolic syndrome, and related diseases.
The most intriguing finding of the initial study was that the BW gain in the mice treated with activated charcoal was significantly lower than in the HFD + Veh group, which was similar to that in the Con group, with no difference in food intake or thyroid function between the HFD + 5%C and HFD + Veh groups. Moreover, acidic activated charcoal significantly improved HFD-induced hyperinsulinemia, insulin resistance, WAT hypertrophy and intestinal length shortening and could reduce cecum volume to levels comparable to the Con group.
In order to validate the findings of the first study and clarify which concentrations of activated charcoal exhibited the highest anti-obesity properties, we investigated progressively higher doses of the material. Our results showed that 4.5% activated charcoal was the most effective in improving the phenotypic changes induced by long-term HFD feeding without any detrimental effects. This information might be applicable to humans in future food additives to prevent obesity.
In the initial study and dose-dependent study, liver TG levels did not increase in the HFD + Veh group. In this study, inulin, dextrin, and raffinose were used as a vehicle to prevent constipation by charcoal. It was documented that inulin could attenuate hepatic TG accumulation in several mouse models, such as high-fat/high-sucrose treated mice and high-cholesterol treated mice (14, 15). Additionally, raffinose could ameliorate hepatic lipid accumulation in high cholic acid treated rats (16). The absence of marked TG accumulation may be associated with the action of inulin and raffinose to attenuate hepatic steatosis.
According to the metabolomic analysis, large amounts of BAs were excreted into the feces by inhibiting their reabsorption in the intestine, and so altered amounts of BAs in the enterohepatic circulation were detected in the liver. As a compensatory response, Cyp7a1, a rate-limiting enzyme for de novo BA synthesis from cholesterol, may have been increased by the charcoal; a sufficient supply of cholesterol is necessary to synthesize BAs. The intestinal absorption of dietary cholesterol requires emulsification by BAs (17). A large amount of BAs were combined with activated charcoal and excreted through the feces, which demonstrated altered digestion and absorption of cholesterol in the intestine. These changes may increase Hmgcs1 and Hmgcr.
Prior studies have found that FXR disruption and activated TGR5 in the intestine, which are closely associated with intestinal BA signaling and nutrient homeostasis, produce an anti-obesity effect and improve glucose metabolism (18–22). The inconsistency of changes in intestinal FXR/TGR5 between the two independent experiments prompted us to conclude that FXR/TGR5 was not a primary reason explaining the anti-obesity effect of acidic activated charcoal. The charcoal also absorbed BAs, cholesterol, and neutral lipids in the small intestine, thereby enhancing the excretion of fat into the feces and preventing BW gain.
It was earlier reported that obese patients were in a state of persistent chronic inflammation, which mediated the development of obesity-related diseases, especially type 2 diabetes (23, 24). In particular, adipose expansion induces the adipose inflammation related to insulin resistance (25–27). Indeed, the expression levels of Il1b and Ccl2 in the HFD + 5%C group were significantly decreased vs. the HFD + Veh group. Although no remarkable differences were observed for Cd68 or Tnf, the HFD + 5%C group showed a decreasing trend. In the dose-dependency experiment, the mRNA levels of Cd68, Ccl2, Tgfb1, and Col1a1 all decreased with increasing levels of acidic activated charcoal. This indicated that the charcoal alleviated the adipose microinflammation caused by obesity, with larger doses of the material being more effective.
Since the activated charcoal was a powder and was not digested by the animals, we were concerned about pneumoconiosis and damage to the digestive tract (28). However, there were no visible differences of the lungs and gastrointestinal tract, and inflammation-related gene levels were not increased in the HFD + 5%C compared with HFD + Veh groups. Based on those findings, we consider the oral administration of acidic activated charcoal to be relatively safe, although further long-term studies are required to confirm its safety and applicability in humans. Moreover, because activated charcoal can absorb lipids, it may disrupt the absorption of fats, fat-soluble vitamins, and folic acid (29–31). As the next step, extended experiments will be mandatory to monitor for deficiencies in essential FAs and fat-soluble vitamins. Acidic activated charcoal can absorb not only lipids, but also toxic contaminants in foods, and metabolites from microbiota, such as indole acetate and lithocholic acid. These metabolites are ligands for the aryl hydrocarbon receptor and pregnane X receptor, which modulate intestinal mucosal immunity (32). It is of great interest whether acidic activated charcoal can modulate the crosstalk between metabolites, the microbiome, epithelial cells, and immune cells.
Conclusion
Acidic activated charcoal improved HFD-induced obesity and insulin resistance without any serious adverse effects. These beneficial effects were likely due to modulating lipid absorption and altered FA/BA metabolism.