Anm53(1)_abstract.indd

Cell-Specific Regulation of TGF-␤1 Promoter Prevention of Amyloid ␤-Induced Memory in Dermal Papilla Cells from Androgenetic Impairment by Fluvastatin, Associated with the Decrease in Amyloid ␤ Accumulation and Oxidative Stress in Amyloid ␤ Injection Mouse Model Department of Regenarative Dermatology, School of Medicine, Osaka University, Osaka, H. Kurinami1, N. Sato1, M. Shinohara1, D. Takeuchi1, S. Takeda1, M. Shimamura2, T. Ogihara3, R. Morishita1 1Department of Clinical Gene Therapy, Graduate School of Aims: We have already reported that TGF-β1 is up-regulated by Medicine, Osaka Universitsy, 2Department of Advanced androgen in dermal papilla cells (DPCs) from androgenetic alopecia Clinical Science and Therapeutics, Graduate School (AGA) using our coculture system of DPCs transiently transfected of Medicine, The University of Tokyo, 3Osaka General with androgen receptor (AR) expression vector and keratinocytes Medical Center, Osaka Prefectural Hospital Organization (KCs). This finding suggested that the regulation of TGF-β1 by androgen is a key step in the pathogenesis of AGA. Our purpose is to Aims: Alzheimer disease (AD), the most common cause of examine the possible regulatory mechanism of TGF-β1 gene expres- dementia in the elderly, is characterized by Aβ-containing plaques and neurofibrillary tangles, synaptic and neuronal loss, along with pro- Methods and Results: We analysed TGF-β1 promoter activity gressive cognitive impairment. Although there are growing evidences of the pTGF-β1-Luc vector, which was constructed by connecting the suggesting the beneficial effects of 3-hydroxy-3-methylglutaryl 5’ upstream region of the TGF-β1 gene (-1362 bp upstream from the coenzyme A reductase inhibitors (statins) on AD, this notion is still transcription start site) to the luciferase reporter gene. When pTGF- controversial. We investigated the efficacy of statins for Aβ-induced β1-Luc and AR expression vector were cotransfected in DPCs from AGA, R1881 increased luciferase activity to around 10-fold. In con- Methods: We employed male ddy mice (6 weeks old), which trast, this induction was not observed in CV-1 cells and transformed were injected Aβ1-40 into the cerebral ventricle. Fluvastatin (5 mg/ DPCs. Thus the regulation of promoter activity by androgen is cell- kg/day) was administered ollary for 2 weeks before Aβ injection, and specific. From analysis of deleted pTGF-β1-Luc vector using DPCs mice were treated for a further 3 weeks. Then water-finding task were from AGA, there are two possible regulatory regions (-1131~-731 and performed to evaluate cognitive function. Biochemical and immuno- -459~-323) and two negative regulatory regions (-1326~-1127 and histochemical analysis were also performed.
-735~-459). The region of -1131~-731 region contains the androgen- Results: The present study demonstrated that pretreatment with responsive consensus region (5’-GCC AGT TGG CGA GAA CAG fluvastatin, but not post-treatment just after Aβ exposure, prevented TTG GCA CGG G).
Aβ-induced memory impairment. We also observed that fluvastatin Conclusions: We suggested the possibility that this region might significantly decreased Aβ accumulation and oxidative stress after Aβ function as a key regulatory site for androgenetic alopecia.
injection. Mice treated with simvastatin did not demonstrate preven-tion of Aβ-induced memory impairment, and showed no significant decrease in oxidative stress. More importantly, fluvastatin signifi-cantly prevented the loss of neurons in the basal forebrain induced by Aβ. Conclusions: The present study demonstrated that fluvastatin significantly prevented memory impairment induced by Aβ. The ben-eficial effects of fluvastatin might be explained by the preservation of neurons through a significant decrease in Aβ accumulation and oxida-tive stress. In clinical practice, the timing of start of treatment with fluvastatin might be critical to acquire a beneficial effect on cognitive function.
2008 S. Karger AG, Basel0250–6807/08/0531–0059$24.50/0 Fax ϩ41 61 306 12 34E-Mail [email protected] [5] Neubauaer M et al., FEBS Lett. 577: 277–283, 2004 [6] Kakudo N et al., Biochem. Biophys. Res. Coommun., 359: 239–244, FGF2 Stimulates Preadipocyte Differentiation [7] Eda H te al., Biochem. Biophys. Res. Coommun.,366: 471-475, 2008 through a Reduction in Intracellular TAZ [8] Hong J.H. et al., Science., 309: 1074–1078, 2005 H. Eda1,2, K. Aoki1, H. Ohtomo3, K. Ohkawa1 1Department of Biochemistry, The Jikei University School of Medicine, Tokyo, Japan, 2Department of Orthopaedic Surgery, Chikuma Central Hospital, Nagano, Japan, 3 Department of Pain Clinic, The Jikei University School of Elegans by an Antioxidant, Platinum Nanoparticle Aims: In recent years, excess visceral fat accumulation or obesity J. Kim1, M. Takahashi2, T. Shimizu2, T. Shirasawa3, has reached epidemic proportions in the developed world, resulting M. Kajita4, A. Kanayama5, Y. Miyamoto1 in an increase in the prevalence of metabolic syndrome. Many stud- 1Department of Integrated Biosciences, University of ies have reported the importance of the rule of adipocytes as endo- Tokyo, Chiba, 2Research Team for Molecular Biomarkers, crine cells for energy metabolism and insulin sensitivity in metabolic Tokyo Metropolitan Institute of Gerontology, Tokyo, syndrome [1][2][3]. Some of the mechanisms of adipocyte differen- 3Department of Aging Control, Graduate School of tiation have been clarified, such as the role of PPARgamma in the Medicine, Juntendo University, Tokyo, 4APt Co., Ltd, transcription mechanism [4]. It is known that fibroblast growth factor Tokyo, 5Department of Global Infectious Diseases and 2 (FGF2) signaling induces the expression of PPARgamma [5] and Tropical Medicine, National Defense Medical College, the relation between FGF2 and the adipogenic differentiation [5] [6], however the detailed mechanism is still unclear. Previously it has been reported that the transcriptional coactiva- Aims: Recently, we have shown that platinum nanoparticles tor with PDZ-binding motif (TAZ) protein acts as a transcriptional (nano-Pt) are a superoxide dismutase (SOD)/catalase mimetic. regulator for the differentiation of mesenchymal stem cells into osteo- Accumulated data indicate that antioxidants are helpful materials to blasts and adipocytes [7]. Additionally we reported that FGF2 reduces extend the Caenorhabditis elegans (C. elegans) lifespan. However, the TAZ protein in osteoblastic cells and that the expression of TAZ the effect of well-known SOD/catalase mimetic EUK-8 on the is involved in osteoblast proliferation and differentiation [8]. Because lifespan is controversial. Thus, further clarification of the effects of preadipocytes and osteoblasts derive from mesenchymal stem cells, SOD/catalase mimetics on lifespan is needed. The present study was this study investigates the detailed mechanism of FGF2 stimulation designed to elucidate the survival benefit conferred by nano-Pt, as Methods: Preadipocyte-like cells, MC3T3-G2/PA6, were cul- Methods: We cultured age-synchronized wild-type N2 and mev- tured with various concentrations of FGF2 or without. Two days 1(kn1) mutant in liquid culture medium with nano-Pt or EUK-8 at later, total mRNA was extracted from these cells and aP2 mRNA 20°C. The mev-1(kn1) mutant has the shortened lifespan due to exces- and osteocalcin mRNA, which are the markers for adipocyte and sive oxidative stress. We counted survivals and transferred them into osteoblast differentiation, respectively, were detected by RT-PCR. fresh medium every other day. In the paraquat assay, we measured the Additionally, TAZ protein levels in these cells were detected using survival of nano-Pt or EUK-8-pretreated worms with 0.4 M paraquat incubation for 12 hrs. Fluorescent intensity of lipofuscin was esti- Results: FGF2 increased aP2 mRNA level, dose dependently mated with 10-day-old worms and detection of ROS with fluorescent and decreased osteocalcin mRNA level in preadipocyte-like cells. probe DCF was performed with nano-Pt or EUK-8-pretreated worms Furthermore, FGF2 significantly decreased intracellular TAZ protein for 2 days from day 5. Thermotolerance assays were performed with levels which act as a corepressor of PPARgamma, an aP2 transcrip- 5-day-old N2 at 35°C for 16 hrs. Moreover, we monitored the effects tional factor, and a coactivator of Runx2, an osteocalcin transcrip- Results: At 0.5 mM, nano-Pt significantly extended the lifespan Conclusions: Our previous report demonstrated that FGF2 of N2 nematodes and at 0.25 and 0.5 mM, nano-Pt recovered the stimulated preosteoblast proliferation but inhibited differentiation shortened lifespan of the mev-1(kn1) mutant. In both instances, and calcification through a decrease in the TAZ protein level [6]. The EUK-8 at 0.05, 0.5, and 5 mM did not extend C. elegans lifespan. present study indicates that FGF2 signaling decreases the intracellular When 0.4 M paraquat was loaded exogenously, nano-Pt (0.1 and 0.5 TAZ protein, and the reduction of TAZ increases aP2 mRNA through mM) and EUK-8 (0.5 and 5 mM) were effective in rescuing worms. activation of PPARgamma. The effects of FGF2 signaling may be a Moreover, 0.5 mM nano-Pt significantly reduced the accumulation of key cause of adipogenic differentiation. These results contribute to lipofuscin and ROS induced by paraquat. We measured the in vitro understanding of the mechanism of adipogenic related deseases.
dose-dependent quenching of O2 and H2O2, indicating that nano-Pt is a more potent SOD/catalase mimetic than EUK-8. Nano-Pt neither increased thermotolerance nor interacted with dietary restriction sig- [1] Hotamisligil G.S. et al., J. Clin. Invest., 95: 2409-2415,1995 [2] Yamauchi T. et al., Nat. Med., 7: 941-946, 2001 Conclusions: These results suggest that exogenously-treated [3] Maeda N et al., Nat. Med., 8: 731-737, 2002 nano-Pt can extend lifespan of C. elegans by reducing oxidative [4] Picard F et al., Nature, 429: 771-776, 2004 8th Scientific Meeting of the Japanese Society of Anti-Aging Medicine stress, regardless of thermotolerance or dietary restriction. Both of Conclusions: In the post-menopausal women, rAI values were nano-Pt and EUK-8 counteract acute oxidative stress such as para- influenced by smoking. In the pre-menopausal women, the subjects quat-induced. Taken together, nano-Pt has interesting anti-aging who engaged in regular exercise had significantly lower rAI values properties.
than the subjects who did not exercise regularly.
Lifestyle Factors, and the Radial Artery Kaoru Fujii1, Ayako Higo1, Hiroshi Hirose1,2, Hirokazu Yokoyama1,2, Takanori Moriki1,2, Hirotaka Shibata2, Kazuko Uno1, Masumi Tominaga2, Goji Hasegawa2, Minako Tsujioka1,2, Hiroshi Kawabe1,2, Ikuo Saito1,2 Michiaki Fukui2, Katsumi Yagi1, Mari Tanigawa1, 1Health Center, 2Department of Internal Medicine, Keio Setusya Fujita1, Toshikazu Yoshikawa3, Naoto Nakamura2 University School of Medicine, Tokyo, Japan 1Louis Pasteur Center for Medical Reserch, 2Department of Endocrinology and Metabolism, Kyoto Prefectural Aims: Augmentation Index (AI) is a parameter that reflects University of Medicine, Graduate School of Medical including aortic compliance, central blood pressure, and cardiac load. Science, 3Department of Gastoroenterological Medicine, The radial artery augmentation index (rAI) is significantly correlated Kyoto Prefectural University of Medicine, Graduate School with the aortic augmentation index (Aortic AI), and has been tenta- tively used as an alternative to the aortic AI clinically. We report here the relationship between rAI measurements and lifestyle factors in Aim: Cytokines and chemokaines are suspected to have a role in the development of impaired glucose tolerance (IGT) and diabetes Methods: The study subjects were 327 women (age: 50 ± 7 mellitus (DM). We aim to characterize the pattern for the status of years). We categorized the subjects into pre-menopausal and post- 3 groups (healthy subjects, IGT patients, and DM patients) through menopausal groups, and then according to smoking status, alcohol an analysis of multiple cytokine and chemokine levels in their blood consumption level, exercise level, and salt intake. We analyzed the plasmarelationships between these factors and rAI. We used the HEM-9000 Methods: We determined 27 cytokines/chemokines (IL-1 β, AI Omron Digital automated sphygmomanometer for rAI measure- IL-1ra, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-12p70, ments and rAI values were adjusted for a pulse rate of 75 beats/min. IL-13, IL-15, IL-17, TNF-α, Eotaxin, FGFbasic, G-CSF, GM-CSF, Although a well-trained nurse conducted the rAI measurements, an IFN-γ, IP-10, MCP-1, MIP-1α, MIP-1β, PDGF-bb, RANTES, VEGF) error pertaining to the sensor position occurred at a frequency of one in human plasma using the beads array. For this study, we included in three people. Both the distance to the reflection point and the reflec- healthy subjects and subjects with IGT, and DM. They were classified tion efficiency affect rAI measurement; thus, before using measured into 3 groups based on the data from complete medical checkups.
values for analysis, variation should be monitored and reproducibility Results: In comparison to healthy subjects, subjects with IGT showed elevated levels of MIP-1α, IL-2, TNF-α, IL-12p70, IL-13, A self-administered questionnaire was used to obtain details IL-6, MIP-1β, Eotaxin, IL-1ra. Where as subjects with DM showed about the subject’s lifestyle, and Excel Eiyo-kun FFQg software elevated levels of IL-2, IL-12p70, MCP-1, Eotaxin, IL-1ra, but (Kenpakusha) was used to calculate salt intake. The Mann-Whitney U-test was used to compare the two groups, and a difference of Discussion: These results indicate that impaired glucose homeo- P < 0.05 was considered statistically significant.
stasis leads to changes of cytokine and chemokine networks, thus Results: Age and pulse pressure were both positively correlated indicating that the production of cytokines/chemokines by immune with rAI, and the post-menopausal group (N = 167) had higher rAI val- cells, adipose tissues, and endothelial cells play important roles in ues than the pre-menopausal group (N = 160). Both total cholesterol the development of IGT and DM. This suggests a kind of feedback and low-density lipoprotein cholesterol were positively correlated loop of causality where the artherogenesis induces inflammation and with rAI in the pre-menopausal group, but not in the post-menopausal cytokine/chemokine production, which in turn advances the arthero- genesis. This process significantly contributes to the development of In the post-menopausal group, rAI values were higher in smok- IGT in to DM. As the multi-cytokine/chemokine analysis revealed ers than in non-smokers (P = 0.0160), but there was no difference that many kinds of cytokines/chemokines are related to the develop-between smokers and non-smokers in the pre-menopausal group, ment of IGT and DM, these results will be useful for further research which suggests that women become more sensitive to tobacco after into preventative intervention. Additionally, analysis of the cytokines/ menopause. In the pre-menopausal group, subjects who engaged in chemokines identified in this paper will be useful as bio-parameters regular exercise had significantly lower rAI values (P = 0.0317) than to know the risk of DM/artherogenesis development. Additionally, the subjects who did not exercise regularly. For alcohol consumption these data will be useful in determineing the risk of the development and salt intake, there was no difference in rAI values between the pre- menopausal and post-menopausal groups.
Shogo Asano, Atsushi Suzuki, Sahoko Sekiguchi, Department of Ant-Aging Dock, Aeba Surgical Hospital, Division of Endocrinology and Metabolism, Department of Internal Medicine, Fujita Health University, Toyoake, Aichi, Japan Aims: This study is aimed to examine the relationship between a habit and functional age and the oxidative stress markers. Aims: Suitable mammalian models for aging are desirable to Methods: The subjects were 104 (52 men, 29-85 years old, 52 study human age-associated pathology. Among these model animals, women, 30-84 years old) who underwent Anti-Aging Dock in this hos- klotho-deficient mouse could generate multiple premature aging-like pital from June 2006 to April 2008. For diagnosis of functional aging, features with hyperphosphatemia. Type III sodium-dependent inor- the muscle age was evaluated by body composition analysis measur- ganic phosphate (Pi) transporter, Pit-1, is a ubiquitous protein. Pit-1 is ing weight bearing index (WBI), blood vessel age by pulse velocity essential to maintain the cell activity, because Pi should be supplied analysis, neurological age by Wisconsin Card Sorting test (WCST), for ATP synthesis. However, accumulating evidences suggest that Pi hormone age (serum IGF-I, DHEA-s), and bone age by DEXA mea- overload from the extracellular millieu would be stress for the cells, suring bone mineral density (BMD). For the risk factors, oxidative because Pi uptake itself might induce the formation of apoptosome, stress (urine 8-OHdG, isoprostane, STAS: serum total anti-oxidative resulting in the apoptosis of the cells. The aim of the present study score, OSPPI: oxidative stress prospective index), mental stress (serum was to investigate the effect of Pit-1 overexpression on the develop- cortisol, DHEA-s/cortisol ratio), and metabolic function. In addition, the anti-aging QOL common questionnaire data were collected from Method: Mouse Pit-1 gene was ubiquitously expressed under the all subjects. They were divided into 2 groups according to good or bad control of CAG promoter in transgenic (Tg) rats, and pronuclear DNA habits in eating, exercise, sleep, smoking, drinking, and mental symp- microinjection into the rat zygotes. All rats were allowed free access toms. The statistical analysis was performed for these parameters.
to tap water and a normal rodent chow. Proteinuria was measured by Results: “Good eating” group tended to be younger in muscle the biurent method on a 24 h urine collection at each time points. age and hormone age. WBI was 7.7% higher (0.821 ± 0.120, P = 0.031) Routine serum chemistries were measured by an automatic analyzer.
and cortisol was 25.0% higher (13.20 ± 5.74μg/dl, P = 0.034). STAS Results: An early onset of cataract development was observed was 13.1% higher only in men (1261 ± 70, P = 0.000). “Good exercise” after 2 to 8 weeks of age. Progressive degeneration of core protein was significantly associated with muscle age (P = 0.010) and hormone of lens occurred in accordance with aging. Skeletal development and age (P = 0.038). BMD was 10.9% higher in men (1.141 ± 0.178g/cm height were normal in Tg compared with WT animals. However, 2, P = 0.043). High sensitive CRP was Ϫ50.7% lower (52.2 ± 45.6μg/later in their life, Tg rats had lower bone mass, analyzed by DXA and dl, P = 0.021) and urine 8-OHdG production was 40.2% higher in μCT, suggesting the development of early-onset osteoporosis. Body women (8.30 ± 4.64ng/kg/hr, P = 0.042). “Bad sleep” (below 6 hours) weight was significantly decreased in Tg compared with WT rats, a was significantly associated with hormone age, decreasing DHEA-s difference that was more pronounced in male compared with female (766 ± 472ng/ml, by Ϫ34.0%, P = 0.030,) and DHEA-s/cortisol ratio animals. Tg rats showed progressive proteinuria associated with (7.05 ± 4.41, by Ϫ39.0%, P = 0.013). “Smoking” was significantly hypoalbuminemia and dyslipidemia, suggesting the development of associated with blood vessel age (P = 0.015), increasing serum homo-nephrotic syndrome. Proteinuria was detected at 3 months of age and cystein (12.51 ± 5.89nmol/ml, by 36.9%, P = 0.006). It significantly glomerular abnormality assessed by transmission electronmicroscopy affected hormone age (P = 0.003) and bone age (P = 0.019) in men, already observed in kidneys of 8 weeks old Tg rats indicating the decreasing IGF-I (151.1 ± 41.5ng/ml, by Ϫ24.6%, P = 0.045), BMD development of nephrotic syndrome. Survival was much shorter in (YAM 86.53 ± 14.71%, by Ϫ9.7%, P = 0.044) and OSPPI (5.9 ± 21.1, Tg rats. Tg rats died because of malnutrition and cachexia between 7 by Ϫ72.3%, P = 0.034). Non-smoking habits positively affect muscle age in women, increasing WBI by 12.9% (0.746 ± 0.069, P = 0.011). In conclusion, results presented in this study suggest that Pit-1 “Bad drinking” (over 30 mg/day alcohol) deteriorated hormone age overexpression induces cellular stress on several organs, resulting in in men (P = 0.007), decreasing IGF-I (161.7 ± 48.2ng/ml, by Ϫ24.8%, the early-onset of aging-related diseases such as cataract and osteopo- P = 0.029), however in women, increasing IGF-I (196.3 ± 47.4ng/ml, rosis. In addition, phosphate overload causes alteration in the integ- by 34.3%, P = 0.051). In both sex, isoprostane generation rate was rity of the glomerular system leading to nephrotic syndrome. increased (3.49 ± 3.94 ng/kg/hr, by 96.0%, P = 0.007). “Bad mental symptoms” were significanly associated with muscle age (P = 0.004) and hormone age(P 0.004), decreasing DHEA-s/cortisol ratio (8.84 ± 5.57, by Ϫ32.6%, P = 0.003) and STAS (1166 ± 95, by Ϫ4.7%, P = 0.013).
Conclusion: Our data indicate that bad habits can affect the func- tional aging partially mediated by oxidative stress. It will be impor-tant that practicants repeat examinations of the statistics of clinical inspection data and establish a scientific evidence for an anti-aging medicine.
8th Scientific Meeting of the Japanese Society of Anti-Aging Medicine which showed greater longevity, fewer side effects and more natural Effects of Cultured Autologus Fibroblast Grafting on the Rejuvenation of Facial Skin Y. Hori1, 4, H. Kagami2, S. Oshima1, K. Fushimi1, A. Hayashi1, S. Nakada3, Y. Watanabe4 1TES Holdings Co. Ltd, 2The Institute of Medical science, University of Tokyo, 3ClINTEXE Clinic, 4Department Deoxyguanosine (8-OHdG) and Periodontitis of Pharmacology and Pharmacotherapy, Nihon T. Komatsu1, A. Sasahara1, Y. Maehata2, Y. Inoue3, A. Miyagi1, MC-il. Lee2, M. Ikeda4 Aim: Although xenogenic collagen and hyaluronic acid have been widely used as soft tissue fillers, they might cause adverse effects Department of Clinical Care Medicine, 1Division of such as allergic reaction and infection. Accordingly, novel fillers that Dentistry for Special Patients, 2Pharmacology and ESR are less allergic and longer lasting have been awaited. Recently, we Laboratories, Kanagawa Dental College, 3Kanagawa have introduced a novel skin rejuvenating treatment using autologous Children Hospital, 4Department of Comprehensive fibroblasts, which would be able to overcome those shortcomings of Dentistry, KDC Yokohama Clinical Training Center, xenogenic fillers. To date, the effect of autologous fibroblast grafting has been evaluated mostly by subjective manner and there is limited information for the objective findings. In this study, we evaluated the Aims: The 8-hydroxy-2’-deoxyguanosine (8-OHdG) is well clinical effects of cultured autologus fibroblast grafting on skin sur- known as a biomarker to evaluate oxidative stress-induced disease face topography using 3D-replica analyzing system. such as Alzheimer disease or early aging. Recently, 8-OHdG has Methods: From January 2007 to April 2008, 30 subjects includ- also been a useful biomarker for assessing status of periodontitis. It ing 8 males and 22 females (between 30 and 66 years old, 50 ± 12 is well known that early aging including rapidly progressive perio- in average of age) have been treated. Before the treatment, both the dontitis occurs in Down syndrome (DS) patients. We had reported general health condition and the presence of infectious diseases were that the generation of reactive oxygen species (ROS) increased in cul- screened. Autologous fibroblasts were obtained from skin biopsies. tured gingival fibroblasts (GF) from DS patients. Thus, the aim of After enzymatic digestion, the primary culture of fibroblasts was this study was to evaluate oxidative stress and periodontitis from DS maintained in Dulbecco’s modified Eagle’s medium (DMEM) sup- plemented with 10% autoserum at 37°C under the presence of 5% Methods: The study group consisted of DS patients aged from CO2 for 6-8 weeks. On the treatment day, the cells were detached 1year to 39 years and systemically healthy subjects (control) aged from the dishes, washed and resuspended in saline. For the injection, from 4 years to 45 years. Informed consent was obtained and care 3x107cells were used and injection was repeated twice every two was taken to ensure that none of these individuals were under any weeks. At one and six months after the injection, the condition of skin medication. The levels of periodontal status were judged from stan- was analyzed using 3D-replica analyzing system. Statistical differ- dard measurements of probing depth (PD), gingival index (GI). The ences were determined by paired-t test. A P value less than 0.05 was salivary levels of 8-OHdG were determined using an enzyme-linked considered statistically significant.
Results and Discussion: Inflammation seen in the injected Results: The mean values of clinical indices, PD and GI, were region was disappeared within a few days after the treatment. The not significantly different between young (Ϲ 12 years) patients with results from silicon 3D-replica analyses showed that our newly devel- DS (DS-1) and controls (C-1; young), adults (from 30 years to 45 oped skin rejuvenating treatment increased skin surface fineness for years) patients with DS (DS-2) and controls (C-2; adults). The sali- 63.3% and reduced the depth of wrinkles by 6.2% at 6 months after vary levels of 8-OHdG in DS-1 and DS-2 were significantly higher treatment. Moreover, these improvements were getting to be better than those in C-l and C-2. and better day by day. And such improvements between age of the Conclusion: These results suggest that the progressive oxidative subjects were not detected any significant differences. In addition, the stress in DS due to increased the value of 8-OHdG in saliva. These improvements of female subjects were about 1.4 times much more redox imbalances may occur to storage ROS in DS. The high oxida- than that seen in male subjects. These results indicate the importance tive stress of DS in saliva may lead to clinical features of DS, espe- of daily skin care such as maintenance of skin moisture. Thus this cially early aging including rapidly progressive periodontitis. newly developed skin rejuvenating treatment might be highly potent for skin anti-ageing. Since the improvement was observed even at 6 month after treatment, the effects of this new treatment will be main-tained over years. Through this treatment, no serious adverse effect has been observed. Although the underlying mechanisms of the effi-cacy are not fully understood, the results from basic study showed ultrastructural changes consistent with new collagen formation at the end of the 3-month treatment period.
Conclusion: This study documents that the efficacy of our newly developed skin rejuvenating treatment was clearly proved, Although the effect of parthenolide in the etiology AGA remained unclearly, it was demonstrated that parthenolide was promising for Tetsuya Tomita, Daiki Morimoto, Hideki Yoshikawa Department of Orthopaedics, Osaka University Graduate School of Medicine 12
Novel SNP Detection as a Genetic Marker
Introduction: There are many patients with androgenic alopecia (AGA). Dihydrotestosterone (DHT) that is one of androgenic hor- mones was considered to be a major cause of AGA. There are many Tomohiko Urano*†, Ken’ichiro Narusawa‡, drugs for AGA, which inhibit transforming testosterone into DHT, Masataka Shiraki§, Takahiko Usui*†, Yasuyoshi Ouchi*, but the etiology of AGA remained unclear.
Toshitaka Nakamura‡, Satoshi Inoue*† It has been well known that feverfew have the potent role of anti- inflammatory and used as folk remedy for attack of fever. Recently, From the *Department of Geriatric Medicine, Graduate the component of feverfew was analyzed and the main component is School of Medicine, The University of Tokyo, Tokyo, clarified to be parthenolide (PTH). It was demonstrated that partheno- Japan, †Department of Anti-Aging Medicine, Graduate lide inhibited the nuclear transition of NF-kB, which was important School of Medicine, The University of Tokyo, Tokyo, transcription factor and promoted osteoclast formation.
Japan, ‡Department of Orthopedic Surgery, University In previous study, we demonstrated that parthenolide was effective of Occupational and Environmental Health, School of for rheumatoid arthritis (RA). Recently patients with RA and AGA Medicine, Kitakyushu, Japan, §Research Institute and who has been administered PTH orally for RA treatment has shown Practice for Involutional Diseases, Nagano, Japan the effect of restoring hair. In this study, we evaluated the effect of PTH for AGA in vivo and in vitro experiments.
Spinal Osteoarthritis is a very common condition in the axial skel- Material and Methods: In vivo experiment, the hair of C3H/ etons of aged people and a major cause of back symptoms. Vertebral He male mouse on their back was shaved at day 0. In control group, osteophytes, endplate sclerosis and intervertebral disc narrowing 0.5%methyl cellulose solution at 10 ml/kg as vehicle was administered are recognized as characteristic features of spinal disc degeneration. p.o and in PTH group, 40mg/kg parthenolide solution was adminis- Recent studies indicate that the appearance of these radiographical tered p.o from day 1 to day 19. The photograph of growing area of features is influenced by genetic factors, physical loading and other hair on the back was taken every other day and measured the ratio of environmental factors. Moreover, spinal osteoarthritis has been shown restore hair area on the back. At day 19, all mice were sacrificed. to have a familial component and in some studies to be influenced by Human follicle dermal papilla cells (HFDPCs) from white male specific genetic risk factors. In the present study, to identify common were purchased and used for in vitro experiments. In control group, genetic variants associated with spinal osteoarthritis,we examine an HFDPCs were cultured with medium and in DHT group HFDPCs association between polymorphisms in bone and cartilage metabo- were cultured with 50 nM DHT. In PTH group, HFDPCs were cul- lism-related genes and radiographic feautures of spinal osteoarthri- tured with 2 μg/ml PTH. Cell proliferation was evaluated with BrdU tis including osteophyte formation, endplate sclerosis and disc space assay. In control, DHT and PTH group, HFDPCs were cultured and nuclear protein was extracted. The amount of NF-kB p65 protein from For this purpose, we evaluated the presence of osteophytes, nuclear protein of cultured cells was measured with the optical den- endplate sclerosis, and narrowing of disk spaces in Japanese post- sity at the wavelength of 450 nm and calibrated by standard protein. menopausal women. The severities of spinal degeneration including Results: In vivo experiment, the hair on the back showed grow- osteophyte formation, endplate sclerosis and disc space narrowing ing faster in PTH group than that in control group and mean ratio of were assessed semi-quantitatively from Th4/5 to L4/5 disc level or restore hair area on the back showed significant differences between from Th4 to L4 vertebrae by using the grading scale of Genant. Then control and PTH group at day 15,17 and 19.
we assessed radiographical spinal osteoarthritis using scoring system. In vitro experiments, there were no significant differences in Briefly, osteophyte formation at a given disc was graded 0-3 degrees, BrdU assay between control and DHT and PTH group. The amount endplate sclerosis at given vertebra was graded 0-2 degrees, and disc of NF-kB p65 protein from nuclear protein was 2.54 ng in control space narrowing was graded 0-1 degrees. Then we defined sum of group, 2.28 ng in PTH group and 4.33 ng in DHT group respectively. each degree from Th4/5 to L4/5 disc level for osteophyte formation Although there were no significant differences between three groups, on anteroposterior radiographs as a score of osteophyte formation. these results suggested that the amount of NF-kB p65 protein from We also defined sum of each degree from Th4 to L4 vertebra for end- nuclear protein was higher tendency in DHT group than that in con- plate sclerosis and that from Th4/5 to L4/5 disc level for disc space narrowing on lateral radiographs as a score of endplate sclerosis and Conclusion: In this study, it was suggested that parthenolide disc narrowing, respectively. The selected SNPs in bone and cartilage have potent role of growing hair in vivo experiment and the amount of metabolism-related genes were extracted from the Assays-on-Demand NF-kB p65 protein in nuclear protein increased in DHT group. These SNP Genotyping Products database (Applied Biosystems, Foster City, results suggested that in the etiology of AGA there were novel path- CA) and genotyped using the TaqMan (Applied Biosystems) poly- merase chain reaction (PCR) method according to the manufacture’s protocol.
8th Scientific Meeting of the Japanese Society of Anti-Aging Medicine In these search, we found the association of single-nucleotide cies. Muscle contraction requires a large amount of ATP in skeletal polymorphism (SNP) in the insulin-like growth factor-1 receptor muscles. The vast majority of such ATP is generated by OXPHOS. (IGF1R) with spinal disc narrowing. We compared those who carried The high rate of energy consumption in skeletal muscles can cause the G allele (GG or GC) with those who did not (CC) in the IGF1R an electron leakage from the electron transfer chain and results in gene at intron 1 (rs11247361). We found that the subjects with the G increased oxidative damages. Accordingly, oxidative stress and dam- allele (GG or GC) were significantly over-represented in the subjects ages observed in mitochondria of skeletal muscles may be attribut- having higher disc narrowing score. We also found that a synony- able to phenotypic changes associated with aging, such as the loss of mous SNP (Q89R) in the LRP5 gene (rs41494349) is significantly skeletal muscle mass and functions. In order to investigate the path-associated with spinal osteophyte formation score and a SNP at 3’ ological significance of oxidative stress in the skeletal muscle, we UTR region in the Wnt-1-induced secreted protein 1 (WISP1) gene generated skeletal muscle-specific manganese superoxide dismutase (rs2929970) is significantly associated with spinal endplate sclerosis (Mn-SOD)-deficient (muscle-Sod2-/-) mice.
score. LRP5 and WISP1 are Wnt/LRP5 signaling molecules.
Methods and Results: Muscle-Sod2-/- mice showed severe These data suggest that IGF1R, LRP5 and WISP1 are genetic disturbance of physical activities, but no atrophic changes in skeletal determinants of bone and cartilage metabolism. SNPs of IGF-I/IGF1R muscle. In histological and histochemical analyses, the mutant mice and Wnt/LRP5 signaling genes will serve to facilitate early diagnosis, showed centralized nuclei in muscle fibers. In addition, we found that treatment and prevention of spinal osteoarthritis.
NADH dehydrogenase (Complex I) and SDH (Complex II) activi-ties in mitochondrial respiratory chain complexes were selectively reduced in muscle-Sod2-/- mice. Particularly, the loss of Complex II activity was remarkable and was hardly detected in skeletal muscle of muscle-Sod2-/- mice while a strong enzymatic activity was detected in those of control mice. Furthermore we showed that the selective Oxidative Stress in Skeletal Muscle Causes loss of enzymatic activity led the reduced ATP synthesis in skeletal Severe Disturbance of Physical Activities muscle. Subsequently, we performed the rescue experiment using EUK-8. EUK-8 is one of the synthetic salen-manganese compounds with the activities of superoxide dismutase, catalase and oxyradical Tetsuro Horie1, Chizuru Tsuda1, Shin Ishikawa1, scavengers. These enzymatic activities confer these compounds the Yoshihiro Noda1, Hirotomo Kuwahara1,2, ability to inactivate superoxide anions and their dismutation products Satoru Kawakami1,4,5, Tomoe Yoshida1, Yusuke Ozawa1, hydrogen peroxide, thereby preventing hydroxyl radical formation. Takuji Shirasawa1,3,5, Takahiko Shimizu1,4,5 We observed that the single intraperitoneally injection of EUK-8 sig- Research Team for Molecular Biomarkers, 1Tokyo nificantly extended the endurance time of muscle-Sod2-/- mice in Metropolitan Institute of Gerontology, 2Department treadmill task even 96 hours after a injection and increase cellular of Orthopedics and 3Department of Ageing Control ATP contents. Therefore, muscle-Sod2-/- mice is a good model to Medicine, Juntendo University Graduate School of evaluate the muscle dysfunction and the declined physical activities Medicine, Applied Biological Chemistry, United Graduate School of Agricultural Science, Tokyo University of Conclusion: We for the first time showed that muscle-Sod2-/- Agriculture and Technology4, Anti-Aging Science, Co., Ltd mice exhibited drastic physical disturbance without muscle atrophy and the impairment due to oxidative stress generated in mitochondria. Furthermore, we demonstrated the efficacy and limitations of anti-oxidant salen-manganese compounds. Subsequent studies with our Aims: The age-related muscle weakness and loss of muscle mice should help to develop novel drugs or new dietary supplements, mass are major contributors to frailty in the elderly. These changes which would contribute to the improvement of physical disturbance cause the loss of independence and impair the quality of life. One as well as the quality of senescent life.
of the factors triggering these age-related pathological conditions is the accumulation of the cellular damages due to reactive oxygen spe- Fax ϩ41 61 306 12 34E-Mail [email protected]

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