Are polyphenols the life extending structures in humic substances?

Transcription

1 Are polyphenols the life extending structures in humic substances? From: J. Berger & R. Sommer, Max-Planck-Institut für Entwicklungsbiologie, Tübingen Nadine Saul 1, Kerstin Pietsch 1, Ralph Menzel 1, Stephen Stürzenbaum 2, and Christian Steinberg 1 1 Humboldt-Universität zu Berlin, Laboratory of Freshwater & Stress Ecology, Berlin, Germany 2 King s College London, Analytical and Environmental Science Division, London SE1 9NH, United Kingdom

2 Longevity

3 Longevity with HS? female + Humic Substance male?

4 survivors (%) The effects of humic substances on C. elegans as exemplified by HuminFeed Lifespan during HuminFeed exposure 0 mg/l 2.5 mg/l HF (0.2 mm DOC) 5 mg/l HF (0.4 mm DOC) 25 mg/l HF (2 mm DOC) days

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6 Tannins: Contrasty polyphenols a) Tannic acid, b) Gallic acid, c) Ellagic acid, d) Catechin

7 Tannins: Contrasty polyphenols Plants synthesize polyphenols (PP) for their defense against fungi, bacteria and parasites the creation of colors to attract insects PP have antioxidative, anti-inflammatory, and antimicrobial capacities PP protect against Cardiovascular and neurodegenerative diseases Cancer Oxidative stress Ageing

8 Tannins: Contrasty polyphenols Tannins showed health-promoting and toxic properties have protein-binding & precipitating capacities are anti-nutritional substances due to precipitation of digestive enzymes binding of nutritional proteins astringent taste

9 Individuals alive (%) Individuals alive (%) Longevity via tannin exposure µm TS 100 µm µm GS 300 µm µm ES 50 µm µm CT 200 µm Days Days

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12 3 proven mechanisms for life extension

13 3 proven mechanisms for life extension Hormesis

14 3 proven mechanisms for life extension Hormesis Calorie Restriction

15 3 proven mechanisms for life extension Hormesis Calorie Restriction Targeted molecular modulation

16 3 proven mechanisms for life extension Hormesis Calorie Restriction Targeted molecular modulation

17 Hormesis = mild stress (e.g. a toxin in low concentrations) with beneficial effects on organisms Beneficial effects = e.g. longer life or enhanced stress resistance Hormesis in C. elegans: Radiation, inorganic & organic substances and heat stress

18 Hormesis

19 Survivors after oxidative stress (%) Survivors after heat stress (%) Mean lifespan (%) Are tannins hormetic? TA * * GA * EA * CT * * * * * * * Concentration (µm)

20 survivors after heat stress (%) And what about HuminFeed? 80 Thermal Stress Resistance Control HuminFeed mm DOC 0.4 mm DOC 4 mm DOC

21 Mean lifespan in days survivors (%) And what about HuminFeed? mg/l 2.5 mg/l HF (0.2 mm DOC) 5 mg/l HF (0.4 mm DOC) 25 mg/l HF (2 mm DOC) days 15, , , ,5 Control 0.2 mm DOC 0.4 mm DOC 2 mm DOC

22 Body length (µm) Offspring per nematode Are there side effects or is it real hormesis? *

23 Offspring after 85h Are there side effects or is it real hormesis?

24 Length alteration (%) Reproduction alteration (%) Are there side effects or is it real hormesis? A1 A6 A1 A6 A1 A6 0.2 mm DOC 0.4 mm DOC 2 mm DOC mm DOC 0.4 mm DOC 2 mm DOC

25 3 proven mechanisms for life extension Hormesis Calorie Restriction Targeted molecular modulation

26 3 proven mechanisms for life extension Hormesis Calorie Restriction Targeted molecular modulation

27 Calorie Restriction Calorie Restriction= Longevity and ageing deceleration by reducing the amount of food to about % of the ad libitum amount Tannins possess antinutritional capacities

28 Calorie Restriction 1989: Start of a Calorie Restriction rhesus monkey experiment They get only 30% of the ad libitum food amount So far: Less age related diseases like cancer, heart disease or diabetes Better brain function Extension of the median lifespan

29 Calorie Restriction 1989: Start of a Calorie Restriction rhesus monkey experiment They get only 30% of the ad libitum food amount Healthy, but hungry ad libitum Calorie Restriction

30 Individuals alive (%) Calorie Restriction in C. elegans 100 no food (A6) 50 ad libitum Days

31 Quantity of offspring related to control (%) Attraction Assay ~ ~ ~ 6 alternating drops of food bacteria (with and without test substance) 1 L1-Larvae per drop ~ ~ ~ Waiting for 96h Counting the offspring per drop TA 100 µm GA 300 µm EA 50 µm CT 200 µm 140 * *

32 Change of pharyngeal pumping (%) Pharyngeal Pumping Frequency * Day 6. Day 9. Day mmmmmmmm0

33 Mean lifespan variation (%) Lifespan of the eat-2 mutant N2 eat TA 100 µm GA 300 µm EA 50 µm CT 200 µm HF 0.4 mm

34 3 proven mechanisms for life extension Hormesis Calorie Restriction Targeted molecular modulation

35 3 proven mechanisms for life extension Hormesis Calorie Restriction Targeted molecular modulation

36 Insulin / IGF-1 (insulin/insulinlike growth factor) pathway Insulin/DAF-28 DAF-2/insulin receptor AGE-1 DAF-18 PDK-1 AKT-1, AKT-2 DAF-16 Translocation in the nucleus and regulation of diverse genes Enhanced stress response & Longevity

37 p38 MAPK pathway

38 TGF-beta pathway TGF-β = Transforming growth factor beta Controls proliferation, cellular differentiation, immunity, apoptosis, lifespan, etc

39 Microarray

40 Microarray & Mutant Strains Results Both, Tannic acid & Huminfeed, modulate 3 known longevity pathways: The TGF-beta signaling The p38 MAPK signaling The Insulin like signaling They seem to modulate the same molecular mechanisms of longevity

41 Do the properties of HF changed after enrichment with hydroxybenzene? Chemical enrichment of HuminFeed by a polycondensation reaction with hydroquinone or benzoquinone: lifespan extension not changed, but increased tolerance to thermal stress delayed onset of reproduction even more growth reduction stronger pumping activity of the pharynx

42 Conclusion Longevity possible via HF & tannins Hormetic action of TA & EA, tendency for HF No clear hints for Calorie Restriction Molecular background: TGF-beta p38 MAPK insulin like signaling Enrichment with hydroxybenzene molecules boost HF action

43 Conclusion Due to numerous overlapping actions, tannins might be the trigger of longevity in humic substances

44 Thank You! Menzel et al. Enrichment of Humic Material with Hydroxybenzene Moieties Intensifies Its Physiological Effects on the Nematode Caenorhabditis elegans. Environ. Sci. Technol. 2011, 45, Saul et al. Diversity of Polyphenol Action in Caenorhabditis elegans: Between Toxicity and Longevity. J. Nat. Prod. 2011, 74,

45 HuminFeed HuminFeed (Humintech GmbH, Düsseldorf, Germany) was made by an alkaline extraction process of highly oxidized lignite Prior to extraction, the raw lignite material was milled and homogenized The humate solution was separated from the humin via sedimentation of the latter

46 Chemical Enrichment with Hydroquinone HuminFeed (1 g) was dissolved in 2.5 ml of distilled water and further diluted in water to a volume of 50 ml the ph was adjusted to ph 7 addition of 500 mg of the monomer hydroquinone in the presence of oxalic acid and formaldehyde stirring for 1 h at 100 C precipitation by the addition of 6 M HCl at ph 1, filtration, and freeze-drying