300mg/dl (HF/HFD) U/kg 1.5mg/kg. Plasminogen activator inhibitor 1 PAI-1

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2 1 PPRaP Plasminogen activator inhibitor 1PI-1 PI-1 PI-1 PI-1 PI-1 PI-1 PI-1 PI-1 PI-1 () 1 PI-1 PI-1 (1) 1 1 PI-1 5mg/kg 1 3mg/dl T 1 PI-1 (HF/HFD).3U/kg 1.5mg/kg T () total PI-1 Gla-osteocalcin TNF-alpha cross-linked -telopeptide of type I collagen (TX) Total PI-1 ELIS kit (Molecular Innovations) ELIS (Morinaga Institute of iological Science) Gla-osteocalcin high-sensitive enzyme immunoassay kit (Takara-bio) E E WakoPure hemical Industries TNF-alpha ELIS kit (R&D Systems) RatLaps EI kit (Immunodiagnostic Systems) (3) %1 7 % m TRPtartrate-resistantacid phosphatase (TRP)/alkaline phosphatase (LP) staining kit (Wako Pure Industry) () PR RNeasy mini-kit (Qiagen) RN StepOne Plus (Life Technologies Japan) PR (5) 3

3 %O 1% 1mg/mlMEM PI-1nM RN PR PI-1nM 1 lizarin red () student t one-way NOV p<.5 StatView, version 5., software (SS Institute) (1) PI-1 PI-1 PI-1 PI-1 PI-1 PI-1 mrn PI-1 mrn PI-1 PI-1 mrn Plasma total PI-1 (ng/ml) Male Female () PI-1 HE PI-1 1T PI-1 -DPI-1 data not shown (mg/cm 3 ) PI-1 WT (mg/cm 3 ) 5 3 PI-1 KO D (mg cm) PI-1 WT PI-1 KO (3) PI-1 RunxOsterixlkaline phosphatase (LP) PI-1 3- PI-1 data not shown PI-1 Ratio of Runx/b-ctin mrn Ratio of Osterix/b-ctin mrn Ratio of LP/b-ctin mrn Diabetes. and Osteoporosis PI-1 WT PI-1 KO

4 () PI-1, PI-1, PI-1 RNKL mrn PI-1 PI-1 WT PI-1KO Relative mrn expression (/GPDH) Runx Osterix LP Vehicle ctive PI-1 ( nm) ctive PI-1 ( nm) () PI-1 PPRaP PI-1, Mineralization (OD 55nm).5. TRP positive cells / bone perimeter (mm -1 ) Ratio of RNKL/b-ctin mrn PI-1 WT PI-1 KO (5) PI-1 PI-1 PI-1 PI-1 Runx Osterix LP 5 PI-1 5 PI-1 PI-1 Ratio of PPRg/b-ctin mrn 1 1 Ratio of ap/b-ctin mrn 1 1 PI-1 WT PI-1 KO (7) PI-1 PI-1 7, lood glucose (% basal value) 1 1 WT KO # # # WT HF/HSD KO HF/HSD 3 9 (min) Plasma Total holesterol (mg/dl) PI-1 WT PI-1 KO HF/HSD

5 () PI-1, PI-1 PI-1, (Osterix, LP) PI-1, D (mg/cm 3 ) Ratio of Osterix/b-ctin mrn HF/HSD (g cm 3 ) D PI-1 WT PI-1 KO HF/HSD HF/HSD Ratio of LP/b-ctin mrn HF/HSD (9) PI-1 RNKL mrn RNKL/OPG ratio TX 9-D PI-1 9-D Ratio of OPG/b-ctin mrn RNKL/OPG ratio HF/HSD HF/HSD Ratio of RNKL/b-ctin mrn D Plasma TX (ng/ml) PI-1 WT PI-1 KO HF/HSD HF/HSD (1) TNF- PI-1 TNF-1 TNF- Osterix mrn LP mrn 1-D Osterix mrn PI-1 WT PI-1 KO HF/HSD R =.1 p<.1 (mg/cm 3 ). 111 D LP mrn WT KO WT HF/HSD KO HF/HSD R =.9 p< R =.57 p= (11) 1 PI-1 PI-1 TNF- PI-1 TNF-1 1 Tamura Y, Kawao N, Yano M, Okada K, Matsuo O, Kaji H. Plasminogen activator inhibitor-1 deficiency ameliorates insulin resistance and hyperlipidemia but not bone loss in obese female mice. Endocrinology. 155:17-17, 1 doi: 1.11/en.13-1

6 Mao L, Tamura Y, Kawao N, Yano M, Okada K, Okumoto K, Kaji H. Influence of diabetic state and vitamin D deficiency on bone repair in female mice. one. 1:1-1, 1. doi: 1.11/j.bone Yano M, Kawao N, Tamura Y, Okada K, Kaji H. novel factor, Tmem17b, induced by activin-like kinase signal promotes the differentiation of myoblasts into osteoblasts. Exp lin Endocrinol Diabet. 1:7-1, 1. article/end ocrj// 1/_EJ13-7/_article Tamura Y, Kaji H. Parathyroid hormone and Wnt signaling. lin alcium. 13 3:7-5. doi: lia1375 Tamura Y, Kawao N, Oakda K, Yano M, Okumoto K, Matsuo O, Kaji H. Plasminogen activator inhibitor-1 is involved in streptozotocin-induced bone loss in female mice. Diabetes. : , 13. doi: 1.337/db Kawao N, Tamura Y, Okumoto K, Yano M, Oakada K, Matsuo O, Kaji H. Plasminogen plays a crucial role in bone repair. J one Miner Res. : , 13. doi: 1.1/jbmr.191. Tamura Y, Yano M, Kawao N, Okumoto K, Ueshima S, Kaji H, Matsuo O. Enzamin ameliorates adipose tissue inflammation with impaired adipocytokine expression and insulin resistance in db/db mice. J Nutr Sci. : 1-1, 13. doi: 3. Mao L, Yano M, Kawao N, Tamura Y, Okada K, Kaji H. Role of matrix metalloproteinase-1 in the MP- inducing osteoblastic differentiation. Endocr J. : , Okada K, Ueshima S, Kawao N, Yano M, Tamura Y, Tanaka M, Sakamoto, Hatano M, rima M, Miyata S, Nagai N, Tokuhisa T, Matsuo O. Lack of both α-antiplasmin and plasminogen activator inhibitor type-1 induces high IgE production. Life Sci. 93: 9-95, 13. doi: 1.11/j.lfs Tamura Y, Murayama T, Minami M, Matsubara T, Yokode M, rai H. Ezetimibe ameliorates early diabetic nephropathy in db/db mice. J theroscler Thromb. 19(7):-1, 1. Role of plasminogen activator inhibitor-1 in the development of insulin resistance and osteoporosis in obese female mice D Tamura Y. Plasminogen activator inhibitor-1 contributes to the pathogenesis of diabetic osteoporosis in female mice. nd Joint Meeting of the International one and Mineral Society/Japanese Society of one and Mineral Metabolism. May 9-June 1, 13 Kobe