A brew for appetite control ?

Bacteria in my colon are making me fat!

Short chain fatty acids, e.g. acetate, propionate, and butyrate, are produced by the fermentation of dietary fibers in the colon. We’ve heard a lot about probiotic good bacteria and their optimal food know as prebiotics. This post examines some studies that demonstrate how the right combination of pre- and probiotics can influence appetite in mice. The ability of two different fermentable carbohydrates (inulin and β- glucan) to effect  body composition and central appetite regulation in high fat fed mice was examined by Arora and coworkers (2012).   

I can’t wait to revamp my microbiotia. I’ll be at a potluck in an hour!

A fermentation of wheat or rye bran with probiotic intestinal bacteria of the Lactobacillus genus may be  a way by producing short chain fatty acids long before the colon.  Perhaps such a brew or vegan yogurt could work in the opposite direction of an appetizer before entering a setting in which one has the potential to eat out of control.  This was my plan as I read the paper.

Methods

Thirty six C57BL/6 male mice were randomized and maintained for 8 weeks on a high fat
diet containing

• 0% (w/w) fermentable carbohydrate,
• 10% (w/w) inulin
• 10% (w/w) β-glucan

Take note: 0% fermentable carbohydrate is the control. The carbohydrate bulk was recreated with cellulose. The mice were essentially eating powdered paper.

Inulin is a polymer of glucose and five member ring fructose.  β-glucan is a polymer of glucose with 1,3 and 1,6 linkages (mushrooms and sea weed) or   β-1,3 and  β-1,4 in grains. (Nakashima 2018).

Fecal and cecal microbial changes were measured using fluorescent in situ hybridization (FISH) For the reader who has never cut open a rodent, the caecum/cecum is a vacillating organ filled with a greenish/ brownish liquid goo. The human equivalent is the appendix. We need to remember that humans are not always the best model organism for rodents.

Note the abundance of intestinal bacteria in the mouse cecum, bold pink line.

• Fecal metabolites were analyzed with  proton nuclear magnetic resonance (¹H NMR)
• Colonic short chain fatty acids were measured by gas chromatography.
• Body composition and hypothalamic neuronal activation were measured using magnetic resonance imaging (MRI) and manganese enhanced MRI (MEMRI), respectively.
• PYY (peptide YY) concentration was determined by radioimmunoassay.  PYY is a saiety hormone secreted by the ileum to proximal colon in humans.
• Adipocyte cell size and number were also measured.

Mice eating a diet rich in β -glucans gained less weight

Figure 2. The effect of inulin and β-glucan supplementation over the 8-week dietary interventional period (a) weekly cumulative body weight gain, n = 12 per group (b) weekly cumulative food intake over the 8 week dietary intervention period, n = 12 per
group. *  p<0.05, **  p<0.01, ***  p<0.001.

It would appear that both β-glucan and inulin suppress weight gain and food intake. β-glucan appears to be a little more effective than inulin in decreasing food intake. The color of the bars really did not come out well. The β-glucan bars are the first in the triplet groupings. The controls are the last.

The really significant changes were seen in the cecum

Superscipt (*) shows the significant difference between HFD-I or HFD-BG vs.  HFD-C.
*   P<0.05,
**    P<0.01,
***  P<0.001.
Superscript (#) shows the significant difference between HFD-I vs. HFD-BG.
#  P<0.05,
## P<0.01.

The β-glucans do not promote cecal growth itself or the  bacteria therein as much as inulin.  This will be revisited in the discussion

The cecal short chain fatty acid profile

Acetate and propionate levels were significantly increased in HFD-BG and HFD I compared with HFD-C, p<0.05

total SCFA  p<0.05
HFD-BG      34.462.5 mmol/mg
HFD-I         27.862.1 mmol/mg
HFD-C         17.764.1 mmol/mg
acetate  p<0.05
HFD-BG      30.262.4 mmol/mg;
HFD-I          25.163.1 mmol/mg
HFD-C          16.763.1 mmol/mg
Propionate  p<0.05
HFD-BG:     3,061.2 mmol/mg
HFD-I          2,060.9 mmol/mg
HFD-C:        0,660.3 mmol/mg, .
butyrate, not significant
HFD-BG        1.160.3 mmol/mg
HFD-I            0.760.2 mmol/mg                                                                                               HFD-C          0.460.1 mmol/mg

β-glucan and inulin in feeding associated brain activity

UT-Health has a nice tutorial on  feeding circuits in the mouse brain.

In Manganese enhanced MRI, Manganese ions (Mn²⁺) enter into the excitable neuronal cells in the brain producing contrast in the MRI images. The appetite centers are located in the hypothalamus.  This figure combines material from the UT website and Arora (2016).

The arcuate nucleus is comprised of neurons expressing orexigenic (neuropeptide Y, agouti related peptide) and anorexigenic (proopiomelanocortin, melanocyte stimulating hormone) peptides, which project into other areas like  VMH and PVN.

The ventromedial hypothalamus responds to gastrointestinal filling.
Periventricular nucleus releases neuropeptide Y (NPY) as well as agouti-related peptide (AGRP).

paraventricular hypothalamic nucleus receives input from NPY-containing neurons in the arcuate nucleus.  It coordinates metabolism and energy intake.

nucleus of solitaries tractus receives baroreceptor (pressure) information from the gut wall.

Arora and coworkers also profiled classes of intestinal bacteria in the feces over the course of the study.  They reviewed the work of others and the role that these bacteria may play in satiety.

Just my opinion

I left out figures on bacteria in rectal contents and the fecal NMR spectra.   How much are the SCFA products of these bacteria absorbed in the colon?  Did any of these short chain fatty acids make their way into the blood of these mice? I wanted to know if there are transporters for short chain fatty acids in the colon since most of the nutrient absorption occurs in the small intestine.  Al-Mosauwi  and coworkers addressed this in a 2016 study.  They used real time (quantitative) PCR to assess the  MCT1 proton mono carboxylate cotransporter thoruut the human GI tract: :   colon > small intestine > stomach.  Immunolocalization of human tissue revealed  MCT1 to be abundant in the basolateral membranes of epithelial cells of the ascending, transverse, and descending colon, but significantly less prevalent in the sigmoid colon.

A diagram of some very common transporters selected from a random cartoon on the Internet and colorized. On the apical side, short chain fatty acids may diffuse through the membrane in the protonated state.  SCFA may also pass through a bicarbonate/anion exchanger.

Arora and coworkers just reported MRI results for the hypothalamus and surrounding regions.  The hypothalamus is a region of the brain  that lacks a very tight blood brain barrier.    This Wikipedia link contains all sorts of interesting information.   See the blood borne signal section in particular.  As interesting as this paper is, the possibility remains that some other product of  inulin and/or β-glucan are responsible for the changes in feeding behavior.  Perhaps these products require access to regions of the brain not protected by the blood brain barrier.

Borş the brew for appetite control?

An Internet search was perform for beta-glucan and lacto fermentation, inspired by the Lactobacilli in the Arora study.  Though not specifically a lacto fermentation,  Borş is a Hungarian fermentation of wheat bran.  The link provides a recipe for fermentation with natural bacteria as well as a starter.  I’m using the starter recipe with a commercial yogurt starter.

My brew, in all of its glory, fermenting by Lactobacilli in my window seal.  After two days it had a very fruity smell, not at all like the gym sock, vinegar, and rotten butter smell I was expecting.  The taste was rather sweet and fruity.  A quick search of PubMed for borş revealed one publication by Pasqualone and coworkers (2018).  I was way off base.  Phenolics, ferulic acid in particular,  esters (fruity notes) , and ” pungent-sour and goat milk-cheese odor notes” at higher temperatures were reported.   I’m also wondering about the role of oxygen tensions for achieving the “pungent-sour and goat milk cheese odor notes.”

Further studies

My suggestion for further studies would be to test a brew made by fermenting inulin or  β-glucan with some of the Lactobacilli and other bacteria Arora (2016) reported being increased in their study.  Give this brew, or vegan “yogurt” to rodents or ourselves as a possible means of appetite control. Silva and coauthors (2020) have written an interesting reveal of how short chain fatty acids from intestinal fermentations contribute to nervous system health. Would consuming a fermented food product rich in SCFA have the same benefits?

References

Al-Mosauwi H, Ryan E, McGrane A, Riveros-Beltran S, Walpole C, Dempsey E, Courtney D, Fearon N, Winter D, Baird A, Stewart G.(2016) Differential protein abundance of a basolateral MCT1 transporter in the human gastrointestinal tract. Cell Biol Int.40(12):1303-1312

Arora T, Loo RL, Anastasovska J, Gibson GR, Tuohy KM, Sharma RK, Swann JR, Deaville ER, Sleeth ML, Thomas EL, Holmes E, Bell JD, Frost G.(2012)Differential effects of two fermentable carbohydrates on central appetite regulation and body composition. PLoS One. 7(8):e43263  free paper

Logsdon AF, Erickson MA, Rhea EM, Salameh TS, Banks WA.(2018)Gut reactions: How the blood-brain barrier connects the microbiome and the brain. Exp Biol Med (Maywood). 243(2):159-165.

Nakashima A, Yamada K, Iwata O, Sugimoto R, Atsuji K, Ogawa T, Ishibashi-Ohgo N, Suzuki K.(2018) β-Glucan in Foods and Its Physiological Functions. J Nutr Sci Vitaminol (Tokyo). 64(1):8-17.

Pasqualone A, Summo C, Laddomada B, Mudura E, Coldea TE. (2018) Effect of processing variables on the physico-chemical characteristics and aroma of borş, a traditional beverage derived from wheat bran. Food Chem. 265:242-252.

Silva, Y. P., Bernardi, A., & Frozza, R. L. (2020). The Role of Short-Chain Fatty Acids From Gut Microbiota in Gut-Brain Communication. Frontiers in endocrinology, 11, 25. PMC free paper