X-Message-Number: 28044 Date: Fri, 16 Jun 2006 11:38:03 -0700 (PDT) From: Doug Skrecky <> Subject: perilla leaf extract inhibits TNF-a [For a variety of reasons, confidence is high that inhibitors of tumour necrosis alpha (TNF-a) can offer a significant reduction in human age associated mortality increases. Luteolin and the structurally related apigenin are two flavones that are responsible for the TNF-a inhibitory effect of a variety of orally administered food materials. Quercetin is another flavone which fails to inhibit TNF-a orally. The difference appears to be due to the higher molecular weight of quercetin reducing its cellular uptake, and thus reducing bioavailability intercellularly.] [The anti-inflammitory effect of oral perilla leaf is due solely to luteolin.] Biol Pharm Bull. 2002 Sep;25(9):1197-202. Luteolin as an anti-inflammatory and anti-allergic constituent of Perilla frutescens. Oral administration of the perilla leaf extract (PLE) to mice inhibits inflammation, allergic response, and tumor necrosis factor-alpha production. We also found that PLE suppressed the tumor necrosis factor-alpha (TNF-alpha) production in vitro. Using the inhibitory activity of TNF-alpha production in vitro as the index for isolation, we searched the active constituents from PLE and isolated luteolin, rosmarinic acid and caffeic acid as active components. Among the isolated compounds, only luteolin showed in vivo activity: inhibition of serum tumor necrosis factor-alpha production, inhibition of arachidonic acid-induced ear edema, inhibition of 12-O-tetradecanoylphorbol-13-acetate-induced ear edema and inhibition of oxazolone-induced allergic edema. These results suggest that luteolin is a genuinely active constituent which is accountable for the oral effects of perilla. [Only luteolin and apigenin are only orally effective TNF-a inhibitors.] Biosci Biotechnol Biochem. 2004 Jan;68(1):119-25. A hydroxyl group of flavonoids affects oral anti-inflammatory activity and inhibition of systemic tumor necrosis factor-alpha production. We previously reported that oral administration of luteolin can inhibit serum tumor necrosis factor (TNF)-alpha production and several inflammatory and allergic models. We investigated here the effect of various flavonoids which resemble luteolin in structure. Lipopolysaccharide (LPS)-induced TNF-alpha production from macrophages was inhibited by treatment with flavone (luteolin, apigenin, and chrysin), flavonol (quercetin and myricetin), flavanonol (taxifolin), and anthocyanidin (cyanidin chloride) in vitro. Most of these, however, did not affect mice when administered orally. Serum TNF-alpha production was inhibited only by luteolin or apigenin, and only luteolin or quercetin inhibited 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced ear edema. These results suggest that the structure of luteolin: 3',4',5,7-tetrahydroxyflavone, is most suitable for the oral anti-inflammatory activity and that existence or disappearance of a hydroxy group may cause a loss of efficiency. [Luteolin dramatically boosts survival from septic shock from 4.1% to 48%.] Am J Respir Crit Care Med. 2002 Mar 15;165(6):818-23. Luteolin reduces lipopolysaccharide-induced lethal toxicity and expression of proinflammatory molecules in mice. Luteolin is a flavonoid that has been shown to reduce proinflammatory molecule expression in vitro. In the present study, we have tested the ability of luteolin to inhibit lipopolysaccharide (LPS)- induced lethal toxicity and proinflammatory molecule expression in vivo. Mice receiving LPS (Salmonella enteriditis LPS, 32 mg/kg, intraperitoneally) exhibited high mortality with only 4.1% of the animals surviving seven days after the LPS challenge. On the contrary, mice that had received luteolin (0.2 mg/kg, intraperitoneally) before LPS showed an increased survival rate with 48% remaining alive on Day 7. To investigate the mechanism by which luteolin affords protection against LPS toxicity we measured intercellular adhesion molecule-1 (ICAM-1) and tumor necrosis factor-alpha (TNF-alpha) production in response to LPS in the presence or absence of luteolin pretreatment. Treatment of animals with LPS increased serum TNF-alpha levels in a time-dependent manner. The increase in peak serum TNF-alpha levels was sensitive to luteolin pretreatment. Luteolin pretreatment also reduced LPS-stimulated ICAM-1 expression in the liver and abolished leukocyte infiltration in the liver and lung. We conclude that luteolin protects against LPS-induced lethal toxicity, possibly by inhibiting proinflammatory molecule (TNF-alpha, ICAM-1) expression in vivo and reducing leukocyte infiltration in tissues. [Although luteolin and apigenin are both antioxidants, their TNF-a inhibitory ability does not derive from free radical scavenging. Although the free-radical theory of aging might be false, some antioxidents might antagonise aging associated mortality by other means.] Microcirculation. 1996 Sep;3(3):279-86. Apigenin inhibits tumor necrosis factor-induced intercellular adhesion molecule-1 upregulation in vivo. OBJECTIVE: Apigenin is a flavonoid that effectively blocks intercellular adhesion molecule-1 (ICAM-1) upregulation and leukocyte adhesion in response to cytokines in vitro. In the present study, we characterized the effects of tumor necrosis factor (TNF) on ICAM-1 expression in different tissues of the rat. We then assessed whether apigenin alters this response. METHODS: ICAM-1 expression was measured under baseline conditions or 5 h after treatment with rTNF. We used 125I-labeled anti-rat ICAM-1 monoclonal antibody (mAb) and an isotype-matched control mAb labeled with 131I to correct for nonspecific accumulation of the binding mAb. Animals were pretreated with either placebo, apigenin, narigenin (a flavonoid without inhibitory effect in vitro), or vehicle. Additional groups of animals were treated with either allopurinol, glutathione, dimethyl-thiourea, or an anti-CD18 monoclonal antibody in order to assess possible actions of flavonoids that were mediated via free radical scavenging or through interference with neutrophil function. RESULTS: Treatment with rTNF resulted in a marked increase in ICAM-1 expression in all organs studied. The magnitude of the response varied in different organs and increases ranged from onefold (lung) to threefold (muscle). Treatment with apigenin blocked ICAM-1 upregulation in organs with low to intermediate responses to rTNF and it significantly attenuated the increased ICAM-1 expression in organs that normally exhibit more marked upregulation. Treatment with narigenin or vehicle did not affect rTNF-induced ICAM-1 upregulation in all tissues studied. Pretreatment with either allopurinol, free radical scavengers, or anti-CD18 monoclonal antibody did not affect the ICAM-1 upregulatory response to rTNF. CONCLUSIONS: TNF-induced ICAM-1 upregulation in vivo effectively is blocked by apigenin through a mechanism that is unrelated to free radical scavenging or leukocyte function. [TNF-a is involved in the death of heart failure patients.] Circulation. 2005 Feb 22;111(7):863-70. Epub 2005 Feb 7. Tumor necrosis factor-alpha receptor 1 is a major predictor of mortality and new-onset heart failure in patients with acute myocardial infarction: the Cytokine-Activation and Long-Term Prognosis in Myocardial Infarction (C-ALPHA) study. BACKGROUND: Tumor necrosis factor alpha-alpha (TNF-alpha) activation is an independent prognostic indicator of mortality in patients with heart failure (HF). Despite the recognition that several TNF family cytokines are elevated during myocardial infarction, their role in predicting subsequent prognosis in these setting remains poorly understood. METHODS AND RESULTS: We performed a systematic evaluation of TNF-alpha and its type 1 and 2 soluble receptors, together with interleukin (IL)-6, IL-1 receptor antagonist, and IL-10, in 184 patients (132 men; mean age, 64+/-12) consecutively admitted for myocardial infarction. We correlated their values to short- and long-term incidence of death and HF (primary outcome). In 10 patients, we also studied the presence of transcardiac gradients for TNF-alpha and its soluble receptors. The control group comprised 45 healthy subjects who were sex and age matched (33 men; mean age, 65+/-6 years) to the patients. All tested cytokines were increased in patients, and no transcardiac or systemic AV difference was found. After a median follow-up of 406 days (range, 346 to 696 days), 24 patients died and 32 developed HF. Univariate analysis showed that all cytokines were related to outcome, whereas after adjustment for baseline and clinical characteristics, sTNFR-1 remained the only independent predictor of death and HF (hazard ratio, 2.9; 95% CI, 1.9 to 3.8, tertile 1 versus 3), together with left ventricular ejection fraction, Killip class, and creatine kinase-MB at peak. CONCLUSIONS: sTNFR-1 is a major short- and long-term predictor of mortality and HF in patients with acute myocardial infarction. [Although TNF-a increases with age, centenarians avoid these increases in lymphocytes. Extreme human longevity appears to require some avoidance of TNF-a in critical subsystems. Humans which do not possess this attribute appear to have no chance at all of reaching 100 years of age.] Gerontology. 2003 May-Jun;49(3):155-60. High circulating levels of tumor necrosis factor-alpha in centenarians are not associated with increased production in T lymphocytes. BACKGROUND: Aging is characterized by increased inflammatory activity reflected by increased plasma levels of proinflammatory cytokines, concomitant with an altered cytokine profile of T lymphocytes. High plasma levels of tumor necrosis factor (TNF)-alpha are strongly associated with morbidity and mortality in elderly humans. However, the cellular source and mechanisms for the increased circulating TNF-alpha levels are unknown. OBJECTIVE: The aim of the present study was to investigate if high plasma levels of TNF-alpha are associated with increased production of TNF-alpha by T lymphocytes in elderly humans. METHODS: TNF-alpha production by CD4+ and CD8+ T lymphocytes was measured by flow cytometry following stimulation with phorbol 12-myristate 13-acetate and ionomycin in 28 young controls, 14, 81-year-olds and 25 centenarians. RESULTS: Plasma levels of TNF-alpha increased with increasing age. An increased percentage and number of T lymphocytes from the 81 year olds expressed TNF-alpha, whereas centenarians did not show this altered TNF-alpha secretion profile. CONCLUSION: T cells may contribute to the elevated levels of plasma TNF-alpha in healthy elderly subjects, whereas other mechanisms are responsible in very old individuals. [TNF-a is highly variable, and has restricted plasma access. Interleukin-6 levels are increased by TNF-a, and appear to offer a more reliable biomarker for TNF-a than a single measurement of TNF-a itself.] Clin Exp Immunol. 2003 Apr;132(1):24-31. Predicting death from tumour necrosis factor-alpha and interleukin-6 in 80-year-old people. Ageing is associated with low-grade inflammation and markers such as IL-6 possess prognostic value. Tumour necrosis-alpha (TNF-alpha) initiates the inflammatory cascade and has been linked to several age-associated disorders. It remains, however, unknown if TNF-alpha is associated with mortality in old populations. The aim of the present study was to investigate if serum levels of TNF-alpha were associated with all-cause mortality independently of interleukin (IL)-6 in a prospective study of 333 relatively healthy 80-year-old people. A Cox regression model was used to explore effects of TNF-alpha and IL-6 on survival in the following 6 years. A total of 133 participants died during this follow-up period. TNF-alpha was associated with mortality in men, but not in women, whereas low-grade elevations in IL-6 were associated strongly with mortality in both sexes. TNF-alpha explained only 7% of the variability in IL-6 and effects of the two cytokines were independent of each other as well as of other traditional risk factors for death [smoking, blood pressure, physical exercise, total cholesterol, co-morbidity, body mass index (BMI) and intake of anti-inflammatory drugs]. These findings indicate that at least in old populations chronic elevated levels of TNF-alpha and IL-6 have different biological functions that trigger age-associated pathology and cause mortality. [Back to perilla leaf. To no great surprise the powerful anticancer effect of perilla leaf has been attributed to luteolin. Please note that artichoke leaf is also a good source of luteolin.] Biol Pharm Bull. 2003 Apr;26(4):560-3. Inhibitory effect of Perilla leaf extract and luteolin on mouse skin tumor promotion. In the present study, the effects of perilla leaf extract (PLE) and luteolin on 7,12-dimethylbenz[a]anthracene (DMBA)- and 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced skin papillomas in mice were investigated. Topical application of PLE prior to TPA treatment in DMBA-initiated mouse skin resulted in a significant reduction in tumor incidence and multiplicity. An even more potent preventive effect was observed with topical application of luteolin, which we previously identified as an antiinflammatory constituent. PLE was dissolved in drinking water at a 0.05% dose and mice ingested it ad libitum; no significant difference was observed in tumor incidence or multiplicity but there was a significant reduction in tumor volume between the PLE-treated and untreated groups. These results suggest that PLE has potent antipromotion activity and ingesting it as a daily food may provide a beneficial chemopreventive effect. Rate This Message: http://www.cryonet.org/cgi-bin/rate.cgi?msg=28044