PreBiotic-Bio

PreBiotic-Bio Formulation Research

The International Scientific Association for Probiotics and Prebiotics (ISAPP)[1] proposed the following update of the prebiotic definition: “a substrate selectively utilised by host microorganisms conferring a health benefit.” This new definition expands the concept of prebiotic to polyunsaturated fatty acids, phytochemicals, and phenolics. The criteria to define prebiotic properties were: i) the impact on gut microbiota, ii) generation of SCFAs, and iii) improvement in lipid and/or glucose metabolism. Since in current medical practice prebiotics are proposed to patients experiencing gut discomfort, Brochot et al., (2019) chose ob/ob mice, a known model of metabolic diseases and gut microbiota dysbiosis.[2]

Recent studies have suggested that plant extracts rich in polyphenols might modulate a dysbiotic gut microbiota in various animal models including mice fed on a fat‐rich diet.[3] [4] Based on this evidence, Brochot et al., conducted a proof‐of‐concept study in ob/ob mice, a well‐known model of gut microbiota dysbiosis (increased Firmicutes to Bacteroidetes ratio) and metabolic disease to evaluate the ability of Porphyra umbilicalis (PU) and Melissa officinalis (MO) extracts to induce changes in a dysbiotic gut microbiota and also to determine whether these changes may be associated with some ameliorations in both lipid and glucose metabolism.[5] It is noteworthy that in clinical practice prebiotics are proposed to patients suffering from gastrointestinal problems possibly due to a gut microbiota dysbiosis. Thus, a good indicator for treatment by a particular prebiotic is its capacity to change an already dysbiotic gut microbiota. 

Porphyra umbilicalis is a potential functional food with a high content of dietary fibres, minerals, and trace elements as well as proteins and lipids that can exert multiple biological activities and has a high antioxidant capacity.[6] Melissa officinalis is known to have multiple pharmacological effects including anxiolytic, antiviral, and antispasmodic activities, as well as to have an impact on mood, memory, and cognition.[7]

Also studied was an extract of Emblica officinalis Gaertn (also known as amla or Phyllanthus emblica Linn). The fruit of E. officinalis Gaertn is a rich source of vitamin C and tannins and has a strong antioxidant activity.[8] [9] It has also been shown to improve glucose and lipid metabolism in both normal subjects and type 2 diabetic patients.[10]

Discussion

The modifications in the gut microbiota induced by PreBiotic-Bio formula were associated with a significant shift of the overall metabolome of the caecum content. In particular, mice treated with PU and those treated with MO displayed higher caecal levels of propionate than control mice. It is also noteworthy that PU induced a substantial reduction in plasma triacylglycerols (TG), which could be mechanistically linked to the observed increase in propionate, as previously reported.[11] The increase in propionate and reduction in TG were associated with a change in hepatic gene expression but not with a modulation of glucose metabolism.

Conclusion

Modifications of the gut microbiota induced by PreBiotic-Bio formula were associated with a significant shift of the overall metabolome of the caecum content.

Porphyra umbilicalis 

Porphyran is a characteristic polysaccharide of Porphyra umbilicalis which contains up to 48%.

Porphyran is a galactose, highly substituted by the 6-O-sulfation of L-galactose units and 6-O-methylation of d-galactose units. Porphyrans have been reported to be hypolipidemic, anti-cancer, and anti-inflammatory in human beings. Porphyran inhibits NO production in macrophages by blocking NF-B activation in the mouse macrophages of RAW264.7 cells that were stimulated with lipopolysaccharides. This may explain some of the anti-inflammatory effects of porphyran. It has been reported that porphyrans have the potential to prevent hyperlipidaemia due to its excellent antioxidant activities in mice. Previous studies have shown that porphyrans inhibited lipid synthesis in HepG2 cells and also decreased apolipoprotein B100 secretion, realizing its hypolipidemic effect. Oral porphyran alleviates liver damage induced by the high-fat diet of ICR mice, implicating the use of porphyran as a dietary hypolipidemic component.[12]

Porphyran is non-toxic on normal cells, although toxic for cancer cells, and induces cell death in a dose-dependent manner. In vitro anti-proliferative activity of crude and purified porphyran, also in a dose-dependent manner, was reported in HT-29 colon cancer cells and AGS gastric cancer cells. The polysaccharide portion of the crude porphyran was thought to account for anti-proliferative activity via apoptosis, as indicated by increased caspase-3 activity.12

Floridoside is a neutral heteroside found in many red seaweeds - up to 42% in Porphyra umbilicalis.

Biological results obtained in a research study strongly indicate a close relationship between floridoside and innate immunity. Floridoside constitutes a potent activator of the classical complement pathway and this activation seems to be mediated by the recruitment and the activation of natural IgM

directed against this molecule. This is the first time that the molecule of floridoside has been described as an activator of the complement system, particularly via the classical pathway.[13]

Phyllanthus emblica

Phyllanthus emblica contains some very potent phytochemicals: Flavonoids: Chebulic acid, chibulinic acid, rutin, leucodelphinidin, Ellagic acid, gallic acid, ethyl gallate, corilagin. Tannins: Gibberellin A-1, gibberellin A-3, gibberellin A-4, gibberellin A-7, gibberellin A-9. Diterpenes: Zeatin, zeatin nucleotide, zeatin riboside.14

Antimicrobial Effect

Aqueous infusion extract of Phyllanthus emblica exhibited potent antimicrobial activity against Enterobacter cloacae followed by Escherichia coli and Klebsiella pneumoniae. Aqueous infusion and decoction of E. officinalis exhibited strong antibacterial activity against E. coli, K. pneumoniae, K. ozaenae, Proteus mirabilis, Pseudomonas aeruginosa, Salmonella typhi, S. paratyphi A, S. paratyphi B and Serratia marcescens. 1,2,4,6-tetra-O-galloyl-β-D-glucose, from Phyllanthus emblica was reported for anti-viral activity against anti-herpes simplex virus in vitro. Antifungal property of E. officinalis was reported against Aspergillus. Fruit ethanol and acetone extracts showed moderate activity against Fusarium equiseti and Candida albicans where Grisofulvin was used as standard antibiotic.[14]

Antidiarrheal Effect

In Ayurvedic medicine, fruit decoction of Phyllanthus emblica is mixed with sour milk and given in cases of dysentery. Infusion of the leaves with fenugreek seed is given for chronic diarrhea. Recent report suggest that the Phyllanthus emblica fruit extract possesses antidiarrheal and spasmolytic activities, mediated possibly through dual blockade of muscarinic receptors and Ca2+ channels, thus explaining its medicinal use in diarrhea.14

Hepatoprotective Effect

Phyllanthus emblica and its flavonoid quercetin were found to be hepatoprotective against acetaminophen-induced liver damage in Albino rats and mice. The research results indicated that the possible mechanism of P. emblica for hepatoprotective activity is in decreasing glutathione depletion and preventing stimulation of cytochrome P450. Since quercetin alone was more effective than the P. emblica extract, it is thought to be the active principle. Toxic effects induced by lead nitrate and aluminium sulphate were also counteracted by the administration of Phyllanthus emblica extract and ascorbic acid in albino rats. It is also reported that Phyllanthus emblica has strong hepatoprotective effect against carbon tetrachloride-induced hepatic damage.14

Immunomodulatory Effect

Laboratory experiment in mice showed that an aqueous Phyllanthus emblica extract natural killer cell activity and antibody-dependent cellular cytotoxicity in mice. Ethanol extract of Phyllanthus emblica showed biphasic activity in ulcerated mice with dose-dependent healing effect. It efficiently reduced pro-inflammatory cytokine, tumour necrosis factor α (TNF-α) and interleukin-1β (IL-1β) levels and appreciably up-regulate the anti- inflammatory cytokine (IL-10) concentration.14

Anti-ulcerogenic activity

The ethanolic extract of Phyllanthus emblica has found highly effective in controlling growth of H. pylori in-vitro with minimum inhibitory control ranging from 0.91 to 1.87 μg/ μl. The result concluded that the plant ethanolic extract is well retained with total phenolics, reducing power, flavonoids and the antioxidant properties which make amla a proper remedial use against H. pylori infection and gastric ulcer.[15]

Melissa officinalis

Phytochemical investigations on Melissa officinalis have revealed the presence of various phytochemicals including terpenes (monoterpenes, sesquiterpenes and triterpenes) and phenolic compounds (phenolic acids, flavonoids and tannins). The main active constituents of Melissa officinalis are volatile compounds (e.g. geranial, neral, citronellal and geraniol), triterpenes (e.g. ursolic acid and oleanolic acid), and phenolics (e.g. cis-and trans-RA isomers, caffeic acid derivatives, luteolin, naringin and hesperidin).[16]

Antimicrobial effects

Melissa officinalis has shown notable antimicrobial effects on Gram-negative pathogenic bacteria, such as Pseudomonas aeruginosa, Salmonella enteritidis, Salmonella typhi, Escherichia coli, and Shigella strains, particularly on the multi-resistant strains. Melissa officinalis showed moderate antimicrobial activity against Candida albicans, Candida glabrata, Candida krusei and Aspergillus fumigatus (fungal strains), and S. aureus, E. coli, P. aeruginosa and Mycobacterium intracellulare (bacterial strains). These findings verify the traditional applications of Melissa officinalis in the treatment of fevers, wounds, scrofula and aphtha.16

Anti-inflammatory

Melissa officinalis has traditionally been used to treat several inflammatory diseases including asthma and joint inflammation. It is also used as a pain killer. Several pharmacological experiments revealed anti-inflammatory and anti-nociceptive effects of the plant. Pre-treatment with aqueous extract of M. officinalis significantly reduced inflammagen-induced paw edema in rats and decreased the nociceptive response. The anti-inflammatory mechanism of Melissa officinalis seems to be related to its citral content which has been reported to inhibit TNF-???? in RAW 264.7 cells stimulated by lipopolysaccharide (LPS), and suppress IL-6 and IL-1???? in LPS-stimulated peritoneal macrophages.16

Antiviral

Antiviral activity Aqueous extracts of Melissa officinalis have been reported to inhibit the development of several viruses. Extracts of Melissa officinalis have been shown to exert significant anti-HSV-1 and anti-HSV-2 activities in vitro. Anti-human immunodeficiency virus type 1 (HIV-1) activity has been reported for an aqueous extract of Melissa officinalis in in vitro studies.16

Antianxiety effect

There are several records on the traditional use of Melissa officinalis as a tranquilizing medicine. In recent decades, several studies have supported the anxiolytic effects of M. officinalis. In an in vitro study of Melissa officinalis and its main component RA showed GABA-T inhibitory activity. In another in vivo research, oral administration of the hydroalcoholic and ethanolic extracts of the plant induced anxiolytic-like effects, possibly through GABA-T inhibition resulting in an increase in brain GABA levels. An open-label pilot study in stressed volunteers with mild-to-moderate anxiety disorders and sleep disturbances showed that oral administration of a patented standardized extract of Melissa officinalis at a dose of 600 mg/day for 15 days significantly reduces anxiety manifestations by 18%, ameliorates anxiety-associated symptoms by 15%, and lowers insomnia by 42%.16

References

[1] Expert consensus document: The International Scientific Association for Probiotics and Prebiotics (ISAPP) consensus statement on the definition and scope of prebiotics. Nat Rev Gastroenterol Hepatol. 2017 Aug;14(8):491-502. doi: 10.1038/nrgastro.2017.75.

[2] Gut microbiota and diabetes: from pathogenesis to therapeutic perspective. Acta Diabetol. 2011 Dec;48(4):257-273. doi: 10.1007/s00592-011-0333-6.

[3] A Two‐Week Treatment with Plant Extracts Changes Gut Microbiota, Caecum Metabolome, and Markers of Lipid Metabolism in ob/ob Mice. Molecular Food and Nutrition. 2019 https://doi.org/10.1002/mnfr.201900403

[4] Reduced obesity, diabetes, and steatosis upon cinnamon and grape pomace are associated with changes in gut microbiota and markers of gut barrier. Am J Physiol Endocrinol Metab. 2018 Apr 1;314(4):E334-E352. doi: 10.1152/ajpendo.00107.2017.

[5] A polyphenol-rich cranberry extract protects from diet-induced obesity, insulin resistance and intestinal inflammation in association with increased Akkermansia spp. population in the gut microbiota of mice. Gut. 2015 Jun;64(6):872-83. doi: 10.1136/gutjnl-2014-307142.

[6] Obesity alters gut microbial ecology. Proc Natl Acad Sci U S A. 2005 Aug 2;102(31):11070-5.

[7] Nutritional and antioxidant properties of different brown and red Spanish edible seaweeds. Food Sci Technol Int. 2010 Oct;16(5):361-70. doi 10.1177/1082013210367049.

[8] Melissa officinalis L. - A review of its traditional uses, phytochemistry and pharmacology. J Ethnopharmacol. 2016 Jul 21;188:204-28. doi: 10.1016/j.jep.2016.05.010.

[9] Ascorbic acid and tannins from Emblica officinalis Gaertn. Fruits--a revisit. J Agric Food Chem. 2009 Jan 14;57(1):220-5. doi: 10.1021/jf802900b.

[10] Composition and biological activities of hydrolyzable tannins of fruits of Phyllanthus emblica. J Agric Food Chem. 2014 Jan 22;62(3):529-41. doi: 10.1021/jf404703k.

[11] Importance of propionate for the repression of hepatic lipogenesis and improvement of insulin sensitivity in high-fat diet-induced obesity. Mol Nutr Food Res. 2016 Dec;60(12):2611-2621. doi: 10.1002/mnfr.201600305.

[12] Liu Z, Gao T, Yang Y, et al. Anti-Cancer Activity of Porphyran and Carrageenan from Red Seaweeds. Molecules. 2019;24(23):4286. Published 2019 Nov 25. doi:10.3390/molecules24234286

[13] Courtois A, Simon-Colin C, Boisset C, et al. Floridoside extracted from the red alga Mastocarpus stellatus is a potent activator of the classical complement pathway. Mar Drugs. 2008;6(3):407–417. doi:10.3390/md20080019

[14] Gaire BP, Subedi L. Phytochemistry, pharmacology and medicinal properties of Phyllanthusm emblica Linn. Chinese Journal of Integrative Medicine, January 2015 DOI: 10.1007/s11655-014-1984-2

[15] Phytochemistry, pharmacological activities and traditional uses of Emblica officinalis: A review.

Hasan et al., International Current Pharmaceutical Journal, January 2016, 5(2): 14-21

http://www.icpjonline.com/documents/Vol5Issue2/02.pdf

[16] Shakeri et al. Melissa officinalis L.– A review of its traditional uses, phytochemistry and pharmacology. Journal of Ethnopharmacology, 2016. http://dx.doi.org/10.1016/j.jep.2016.05.010