Re: 버드나무 추출물 아스피린의 수명연장기전

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beyond reason

버드나무 껍질에서 추출한 아스피린

- 장수유전자를 켠다. 

The multiple faces of aspirin

1) antioxidant

2) antiinflammatory

3) inhibitor of glycation

aspirin may affect oxidant production and cytokine responses, and block glycooxidation reactions, posing it as a triple threat against the symptoms of aging

aspirin may have a indirect impact on the endogenous production of oxidants by acetylating proteins at the -amino group, thereby preventing interand intra-protein cross-links as well as glycation reactions

최근의 과학자들은 inflammaging(염증성 노화)라는 말에 주목하고 있다.

저용량 아스피린 100mg 

아스피린 경구복용후 30-40분만에 혈중농도가 최대치로 올라가고 100mg이하 소량일때 혈중반감기는 15-20분에 불과하고  항혈소판 효과는 아스피린과 접촉한 혈소판의 cox1은 비가역적으로 아세틸화되어  오랫동안 기능이 억제되기 때문에 항노화목적으로 사용할때는 소량으로 충분하다.  아스피린 반감기는 이처럼 농도의존적이어서 300~650 mg 일 경우 3.1 - 3.2시간, 1g일 경우 6시간, 2g일 경우 9시간이다.

아래의 버드나무 껍질 추출물 800mg안에 15%의 살리신(아스피린)이 들어 있으므로 대략 100mg이다. 

Lifelong Aspirin Supplementation as a Means to Extending Life Span.pdf

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Lifelong Aspirin Supplementation as a Means to Extending Life Span

TRACEY PHILLIPS and CHRISTIAAN LEEUWENBURGH

ABSTRACT

Arising from an initiative by the National Institute of Aging (NIA) requesting novel proposals challenged with increasing lifespan and longevity, our laboratory has generated a hypothesis to test the efficacy of lifelong, low-dosage aspirin administration as a means to achieving this goal. The intervention testing program (currently underway) proposing aspirin as an anti-aging agent evolved from the multitude of properties encompassed in aspirin and the potential of these attributes to prevent the cellular and functional declines, particularly from inflammatory and oxidative sources, evidenced to contribute to aging. Aspirin is a widely administered, cheap, anti-inflammatory, and antioxidant compound that has a variety of positive effects on the immune system and cardiovascular health.

Notably, aspirin may affect oxidant production, cytokine responses, and block glycooxidation reactions, thus posing it as a triple threat against the symptoms of aging. Whether aging is molded by interplay between oxidative stress and inflammatory mediators has received little attention; however, we and other laboratories have explored this notion and have observed an elevated inflammatory status with age. Stemming from these observations and in view of the limited success of antioxidant therapies in improving lifespan in long-lived species, in this article we propose a protocol to examine life-long use of a very low dose anti-inflammatory compound such as aspirin to engage the inflammatory and endogenous oxidative insults accompanying aging and, in so doing, attempt to increase maximum and mean life span.

INTRODUCTION

IN RESPONSE to the National Institute of Aging’s (NIA) call for novel proposals challenged with increasing lifespan and longevity,1 our laboratory generated a hypothesis to test the efficacy of lifelong, low-dosage aspirin administration as a means to achieving this goal. “This project is one of several studies currently being undertaken under the auspices of the NIA Interventions Testing Program (ITP) to test compounds in mouse longevity.”2 The premise for selecting this common place ‘wonder-drug’ as an anti-aging agent evolved from the multitude of properties encompassed in aspirin and the potential of these attributes to prevent the cellular and functional declines, particularly from inflammatory and oxidative sources and their interplay, evidenced to contribute to aging.

One of the primary causes of aging is arguably derived from free radical (oxidants) damage to biological molecules and many attempts have been made to attenuate free radical reactions through supplementation with exogenous antioxidants. By abating these harmful reactions, scientists theorized that aging-associated diseases and the aging process itself would be slowed, ultimately extending life span. However, sizeable experimental evidence now strongly casts doubt over whether any antioxidant is able to prolong life in longlived species such as rodents and humans. Previous studies3–6 have demonstrated the inability of antioxidants in extending the maximum life span in longer-lived species. A selection of these interventions is reported in Table 1, with their effect on mean and maximum life span in short-lived and longer-lived species.

While these studies provide evidence that shorterlived species may benefit from antioxidant therapy, there is no evidence that longer-lived species benefit from the addition of antioxidants or supplements containing antioxidant. Although it may be scientifically interesting and relevant to extend the life span of a worm from 2 weeks to 4 weeks, it becomes clear that testing must be performed in longer living species to truly indicate a more definitive cause and effect interaction. Moreover, studies of antioxidant supplementation often have one important shortcoming: the lack of food intake data. Without monitoring food intake, it is impossible to determine to what extent some of the increases in life span are due to the effect of calorie restriction, rather than just the antioxidant. In studies where food intake was monitored, there was an associated decrease in body weight of up to 10–15% with the intake of various antioxidants.3–6 Thus, whether or not beneficial effects are due to antioxidant supplementation or to calorie restriction remains to be eval‎uated. Moreover, adding antioxidants does not guarantee alterations in the endogenous production of oxidants and could negatively affect redox balance and therefore protein expression‎. Endogenous hydrogen peroxide production is a potent stimulus for gene activation.7,8 Hence, beneficial effects of the antioxidants could be annulled by the negative effect on cell signaling. Considering the limited effectiveness of antioxidant supplementation to prolong lifespan in longer-lived species, an alternative approach still focused on minimizing the “endogenous” free radical damage and inflammation to biological molecules becomes necessary. Aspirin supplementation may present as a strong candidate to fill this void and concurrently tackle other age-linked deteriorations that occur.

The multiple faces of aspirin: antioxidant, antiinflammatory, and an inhibitor of glycation

Aspirin (acetylsalicylic acid) is a widely administered, cheap, anti-inflammatory, and antioxidant compound that is easily dissolved in drinking water or supplemented into food pellets. Although side effects at doses of 75 and 81 mg in humans have been reported, the incidence is low.9–13 Furthermore, aspirin has already been shown to have a variety of positive effects on the immune system and on cardiovascular health, both in rodents and humans.14 Most importantly, aspirin may affect oxidant production and cytokine responses, and block glycooxidation reactions, posing it as a triple threat against the symptoms of aging. A recently emerging and potentially important area of aging research closely associated with free radical biology and redox flux is inflammation. Specifically, key mediators of inflammatory pathways—tumor necrosis factor–alpha (TNF-
) and nuclear factor–kappa B (NF-B)—are under investigation by laboratories (including ours) with regard to a possible role in aging.5,15–19 Inflammatory reactions are an extremely complex series of physiological reactionary events that minimize cellular trauma and promote repair. However, TNF-
and NF-B, which are responsive to multiple stimuli that include oxidative stress, have the ability to have a detrimental effect on cell function. Persistent, unwarranted immunological episodes implicating these and other pivotal inflammatory agents (e.g., inhibitor of B kinase (IKK) complex and inhibitor of B (IB) proteins) have been observed with increased frequency in aged tissues.15,16,20,21

Furthermore, many of the inflammatory cell types are sensitive to reactive oxygen species and regulate their function based on the presence of oxidants.22,23 This presents the possibility that a pro-inflammatory state may become more chronic in association with advancing age. The responsiveness of NF-B to oxidative stress has been well documented23,24; but whether aging is molded by an interplay between oxidative stress and inflammatory mediators has received much less attention Several laboratories have explored this notion through use of lifelong calorie restriction, which reduced the overall chronic inflammation present in rodents.15,16 These researchers formulated the inflammation theory of aging, which complements the free radical oxidative stress theory of aging and the glycooxidation theory of aging. Their postulate16 centers on the increased signaling to NF-B accompanying aging and the ability of calorie restriction to counter this occurrence. Work from our laboratory unveiled a similar response. We assessed TNF-
and several of the signaling proteins intrinsic to TNF-
activation of NF-B. We observed an age-associated increase in TNF-
, which calorie restriction attenuated, and other proteins in the signaling pathway responded similarly. Thus, based on our observations and the finding of others,15,16 an alternative intervention to antioxidant supplementation may be life-long use of a very low dose of a anti-inflammatory compound such as aspirin to extend life span.

Aspirin as a therapeutic intervention to reduce inflammation

Aspirin belongs to the non-steroidal anti-inflammatory drug family of pharmacological agents.25 With its anti-inflammatory, antithrombotic, anti-pyretic, analgesic,25 and antioxidant properties,9 aspirin offers a highly attractive therapy that may prevent an overshoot in the pro-inflammatory component while simultaneously reducing levels of oxidative stress. Aspirin partly elicits its effects by targeting enzymes cyclooygenase I and cyclooygenase II (COX I and COX II). These COX enzymes synthesize biologically active prostaglandins through enzymatic oxidation of arachidonic acid, a fatty acid within cell membranes. The prostaglandins participate in a host of physiological processes in the body and are key players in promoting the inflammatory response.26 One mode of action of aspirin is to intervene in this cascade at the level of cyclooxygenase activity by acetylation of a serine residue at the active site of the cyclooxygenases I and II, thereby inactivating the enzymes.27 Furthermore, aspirin has been reported to inhibit the activation of NF-B through stabilization of IB.28 Therapeutic doses of aspirin suppress lipopolysaccharide-inducible NF-B binding to an NF-B binding site in the TNF-
promoter, lipopolysaccharide-induced TNF-
mRNA accumulation, and protein secretion.28 Other data show that aspirin inhibits NF-kappa B mobilization, induction of VCAM-1 and Eselectin, and subsequent monocyte adhesion in endothelial cells stimulated by TNF-
, thereby providing an additional mechanism for therapeutic effects of aspirin.10 Aspirin also inhibits the activation of TNF-
and NF-B induced by silica, as demonstrated by electrophoretic mobility shift assay.9 Furthermore, thromboxane A2 (THA2) a product of platelet prostaglandin synthesis and a potent vasoconstrictor and clotting agent, also experiences a decline in the presence of aspirin, arising from aspirins’ upstream actions on the active site of cyclooxygenase and a subsequent reduction in the prostaglandins required for THA2 formation.26

Aspirin’s antioxidative actions and other potential benefits of aspirin

In the case of oxidative insult, aspirin may have a indirect impact on the endogenous production of oxidants by acetylating proteins at the -amino group, thereby preventing interand intra-protein cross-links as well as glycation reactions.9–11,29 Furthermore, diabetic rats with a diet supplemented with aspirin were found to have reduced levels of oxidative stress.11 This study indicated that dietary supplementation of the diabetic group with acetylsalicylic acid (100 mg kg
1 body weight day
1) had a marked effect on the level of protein modification (dityrosine and sulfhydryl groups) induced in diabetic crystallins. Effects of aspirin on the pathogenesis of fatty liver, cirrhosis and hepatocarcinogenesis caused by a choline-deficient L–amino acid–defined (CDAA) diet were also examined in male Fischer 344 rats fed a CDAA diet supplemented with 0%, 0.1%, 0.2%, 0.4%, or 0.8% aspirin for 30 weeks.30 Aspirin at concentrations of 0.2% prevented the development of both cirrhosis and preneoplastic and neoplastic nodules, but without any directly associated prevention of fatty changes. Aspirin also prevented hepatocyte proliferation and the generation of thiobarbituric acid-reactive substances and 8-hydroxydeoxyguanosine caused by feeding the CDAA diet. This was analyzed respectively, after 1, 12, and 24 weeks. The results clearly indicated that aspirin, which was not a lipotropic factor, was able to prevent the pathogenesis of cirrhosis and hepatocarcinogenesis caused by a CDAA diet. This prevention may have been partly attributable to the interruption of reactive oxygen species production.30 Hence, long-term low dose aspirin may have a dramatic effect on tumor formation, a pathology often associated with death in rodents. Aspirin in small doses is now recommended by most physicians as a means to lowering the risk of non-fatal myocardial infarction and stroke, and reducing the risk in people with a predisposition of developing vascular disease.31 Moreover, aspirin use is associated with a reduced risk for colon cancer.32 The first clinical trials eval‎uating the efficacy of aspirin and other chemotherapeutic agents in preventing colorectal polyps in people with familial adenomatous polyposis are underway.33 The effectiveness of aspirin in this context is thought to be in the maintenance of the wingless-type (Wnt) signaling transduction pathway. The wnt signaling pathway regulates the presence of -catenin, a protein involved with the promotion of gene transcription, and deemed to be central in the development of colonic neoplasia when the regulatory systems go awry. Aspirin is thought to counter this process due mainly to its cyclooxygenase-inhibitory properties, which result in suppression of tumor cell growth, enhanced apoptosis, and inhibition of angiogenesis.34,35 Chronic use of aspirin by patients to reduce the symptoms of arthritis may also decrease the risk of developing Alzheimer’s disease by inhibiting amyloid-beta aggregation but further investigations to explore this outcome are required before this can be confirmed.36,37 In the most recent clinical trials that have administered NSAIDs as a preventative tool against the risk of dementia the results have been negative.38 However, others have reported some protection against the development of dementia with NSAID use, illustrating the need for further inquiry.36 Furthermore, the rationale for using aspirin as an intervention for aging, as opposed to using other comparable NSAID pharmaceuticals (e.g., ibuprofen), stems from recent studies9–11,39,40 documenting previously unrecognized mechanisms of aspirin action, especially its role as a propagator of anti-inflammatory lipids (termed aspirin-triggered-lipoxins). Recently, it was suggested that aspirin’s mechanisms for blocking polymorphonuclear neutrophil accumulation and infiltration at sites of inflammation was partly due to aspirin-triggered lipoxins and the glucocorticoid-induced annexin 1–derived peptides that are both generated in vivo and act at the lipoxin A4 receptor to halt neutrophil diapedesis.41 Additionally, aspirin can induce hypothermia42 and therefore may alter metabolism and have an effect on the mitochondrial free radical leak. Hence, subtle long-term effects could have a profound impact on longevity and require immediate investigation and resolution.

Dosing considerations

Although the public use this compound extensively, very little is known regarding the effects of chronic long-term use of aspirin on disease pathology and longevity. Moreover, to obtain the anti-inflammatory actions of aspirin sought in the treatment of inflammatory diseases (e.g., rheumatoid arthritis), high dosages of approximately 1 g daily and above have traditionally been prescribed.43 Such regimens, however, revealed one of the potential side effects of aspirin use; a heightened occurrence of stomach irritation and ulceration caused by the blockage of stomach prostaglandins that are essential in regulating overproduction of acid and synthesizing the mucus barrier that prevents its self-digestion. Studies show that acute supplementation using very high doses of aspirin causes mucosal damage.

To circumvent this, lower dosages of aspirin have now been shown to have similar anti-inflammatory and other beneficial biological changes, but without the side effects. Few studies have investigated and compared different anti-inflammatory substances together. One study investigated the relative anti-inflammatory potencies of aspirin, phenylbutazone, indomethacin, three fenamates and several other nonsteroidal anti-inflammatory agents in several rodent models of acute and chronic inflammation.44 Relative toxicities and ulcerogenicities were determined in rats of the same source, strain and sex. The acute ulcerogenic assay measures the minimal irritation potential of these agents and leads to a sensitive index of the safety of such compounds when compared with their therapeutic potencies.44 This study suggests that there was minimal mucosal damage and no significant differences between the various compounds used. Only very high doses of aspirin in rats, supplemented acutely by intragastric dose of 3000 mg/kg/daily, have been shown to cause mucosal lesions.45 One study shows that aspirin (two intragastric doses of aspirin (1.4 mmol/ kg body wt) markedly reduced mucosal prostanoid concentrations in rats, without apparent effects on gastric injury.46 Other studies show that aspirin-induced gastric injury is reduced when aspirin is administered along with phosphatidylcholine.45 Few studies have been conducted where aspirin is supplemented to animals for a longer time. Studies have used doses ranging from as low as 1 mg/kg/day for 110 days47 to 300 mg/ kg/day for periods up to 5 months.48 The absence of more direction in the literature pertaining to the use of aspirin for longer durations makes it difficult to determine the optimal dose for animals. We anticipate that using several low doses of 0.2–3 mg/kg/day either in drinking water or fortified in food compares well with the human clinical trials doses used (low doses use of 81 mg), which show no clinical side effects. In humans, several supplementation regimens have been adopted, including daily, alternating days and single daily dose (325 or 750 mg) schedules,49–52 all of which generally result in an approximate average dose of 5–10 mg/kg/day. However, recent studies suggest that 75–150 mg works just as well, with less chance of internal bleeding.53,54 In the United States, baby aspirin, also available as low-dose adult aspirin, contains 81 mg. In Europe, aspirin comes in doses of 75 and 300 mg.54 Based on the available evidence, a daily dose of aspirin in the range of 75–150 mg is advised for the long-term prevention of serious vascular events in high-risk patients. Few clinical studies have been conducted using aspirin dosages of 325–750 mg (5–10 mg/kg/day) administered at different time intervals.50–52,55,56 However, these dosages have demonstrated positive outcomes on specific cytokines (e.g., reducing C-reactive protein and IL-6 levels) and reductions in the incidence of myocardial infarcts,57–59 providing suggestive evidence that aspirin can modulate the “inflammation theory of aging.” Another consideration pertains to the initiation point of aspirin supplementation. Our study will examine lifelong aspirin use, yet the advantages from an administration protocol embarked on, for example, later in life, lie unknown and need to be investigated. Aspirin has been relied upon on innumerable occasions in the acute sense to combat conditions such as unstable angina51 and to lower C-reactive protein.60 However, the extent to which these episodes of aspirin use affect the individual in the long term is undefined and embodies another area deserving further investigation. 

PROPOSED INTERVENTION PROTOCOL

Although some of the beneficial effects of aspirin are dependent on a (relatively) high dose, we propose to use various low doses of aspirin in this initial study. The major reason for this is that we believe providing a chronic high dose of any compound during the life span of an animal may provide a negative physiological and biochemical effect. Moreover, the doses being used reflect the doses currently utilized by the human population. We realize, that after these initial experiments, future research need to be conducted to further test potential beneficial effects of higher doses and monitor long-term safety and efficacy. Hence, the current study and experimental design for the National Institute on Aging has some of the following features: (1) The primary endpoint is increased longevity. (2) Group size will be sufficient to detect 11% change in longevity at 80% power and 13% change at 90% power. (3) Test subjects will be genetically heterogeneous mice, bred as the four-way cross between CByB6F1 females and C3D2F1 males. (4) Tests will be conducted simultaneously at three sites—the University of Michigan, the Jackson Laboratory, and the NIA Intramural Program. (5) The design will allow comparison of multiple experimental agents with two control groups (ad libitum fed animals and calorie-restricted animals). (6) Two dosages are to be administered: 0.5 and 3 mg/kg/day in the drinking water. (7) The mice will also be tested for a number of age-sensitive traits, such as T cell subsets, cataract development, humoral immunity, cognitive abilities, and spontaneous activity. (8) Agents that show promise in the initial screen will be tested in a follow-up protocol including additional age-sensitive traits as well as detailed pathology. (9) Extensive biochemical analysis (inflammation markers, oxidative stress, signaling proteins related to inflammation) will be performed on plasma and selected organs. (10) All findings—positive or negative—will be submitted for publication. There are striking similarities and discrepancies between the various age-associated diseases between mice and humans. It is well known that rodent’s predominately die from cancer, and humans suffer predominantly from atherosclerosis, with more than 50% dying from cardiovascular disease. In addition, it is well-established that cardiovascular disease is strongly influenced by factors such as diet, exercise, and stress and that very old humans (85 years) often die from conditions such as cancer and neurodegenerative diseases. Hence, depending on the model used (humans, animals) and environmental changes (exercise, diet, stress) aspirin’s action could have different effects at different times during the life span of an organism. We are interested in preventing “normal aging,” and very old humans do not die from cardiovascular disease, which we believe is largely a “preventable disease.” We are attempting to capitalize on aspirin’s other therapeutic qualities in conjunction with its well-established abilities as an anti-inflammatory, antioxidant to achieve a multifaceted approach against the varied systemic degeneration characteristic of aging.

CONCLUSION

In spite of the fact that humans use aspirin on a massive scale worldwide, no long-term rodent (life-long) and human (20–30 years) prospective randomized clinical studies have been conducted to determine its effect on life span. Moreover, aspirin’s use, as opposed to other similar pharmaceuticals (e.g., ibuprofen), stems from potential additional benefits derived from aspirin. These include some of the following: propagation of anti-inflammatory lipids, aspirin-triggered-lipoxins, reduction in pro-inflammatory cytokines, and prevention of glycation and post-translational modifications to protein.39,40 These effects may give support to the argument that aspirin may provide a greater range of beneficial effects than other similar agents (i.e., statins, ibuprofen). This should also be examined and information on their individual effects would be of great interest to clinicians, health officials and the general population. The proposed studies of the Interventions Testing Program (ITP) will be the first to investigate the potential benefits of aspirin following long-term use and its efficacy to extend maximum and/or mean life span. Furthermore, this proposal also hopes to generate a contender for the Methuselah Mouse Prize, an enticing gauntlet laid down by the Methuselah Foundation under the direction of Aubrey de Grey, to encourage aging research and the discovery of novel ways to control aging processes.61,62 It is a contest designed to accelerate progress towards real longevity-enhancing medicine, and to promote public interest and involvement in research on healthy life extension. To this end, aspirin is an alternative to antioxidant therapy and could potentially be an easy available compound to extend healthy life span in humans. Life-long calorie restriction may always remain the most robust anti-aging intervention; however, lifelong treatment with aspirin may be more easily achieved and is worthy of further investigation. Thus, the question of whether life-long aspirin supplementation can perform such a daunting role is an intriguing prospect and will be resolved sooner rather than later.

[teres major]

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