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2 動脈硬化 び 血漿 ア ポ 蛋 白 濃 度 は そ れ ぞ れ, TG 136.2±6.8, HDL-Ch ±5.2, apo A-II 31.0±1.4, C-II 3.8±0.3, apo mg/100mlで ±0.36%で C-III あ っ た. あ っ た. Ch 58.6±4.0, apo Vol. 7.6±0.4, apo B-48比 の 性 差 も検 討 し た が, apo 210.4±14.1, あ り, 特 にIIb型 討 す る と, B-48比 IV型 は, 2.34 齢 と の 間 に は 有 意 な 相 関 は 認 め な か っ た. February 1986 ml)で E 4.2±0.2 B-48比 6 型3例(Ch A-I B 106.6±9.1, に お い て, No ±10.4, apo VLDLのB-48比 C群 13 IIb型, と年 は, で は4.77±1.48%で IV型 と も に, 有 意 な(0<0.05)増 TG 330.3±65.1mg/100 お よ びIV型 IIb型 で は, に つ い て検 3.67±0.57%, あ り, 症 例 数 は 少 な い が, C群 に 比 して, 加 を認 め た(Fig. B-48比 の 3). 以 上 の 結 果 よ り, 高 脂 血 症 患 者 に お け るB-48 ま た, 有意差は認 め なか っ た. 次 に, 原 発 性 高 脂 血 症 患 者(HLP)群14例 け るB-48比 (Ch を 検 討 し た. IIa型2例, IIb型9例 280.3±11.2, Fig. 1 Fig. 2 にお TG 334.2±442mg/100ml), IV Electrophoretic pattern of apo B subclasses in plasma VLDL analyzed by gradient (3-15%) polyacrylamide gel electrophoresis in 0.1% SDS. Densitometric pattern of apo B subclasses in plasma VLDL analyzed by gradient (3-15%) polyacrylamide gel electrophoresis in 0.1% SDS Fig. 3 Percentage of apo B-48 in total VLDL apo B prepared from normal control and patients with type IIb and IV hyperlipoproteinemia. Fig.4 Correlationbetween percentageof apo B-48 in totalvldl apo B and plasma triglyceride concentrations.
3 Fig. 5 Correlation between percentage of apo B-48 in total VLDL apo B and plasma apo E concentrations. Fig. 6 Correlation between percentage of apo B-48 in total VLDL apo B and plasma
4 Table 1 Clinical data and plasma concentrations of lipids and apolipoproteins in normal control and the diabetics Fig. 7 Percentage of apo B-48 in total VLDL apo B prepared from normal control and the diabetics (DM).
5
6 1) Kane, J. P., Hardman, D. A. and Paulus, H. A.: Heterogeneity of apolipoprotein B:Isolation of a new species from human chylomicrons. Proc. Natl. Acad. Sci. U.S.A., 77: (1980). dodecyl sulfate gel electrophoresis system. Biochem. Biophys. Acta, 446: (1976). 10) Aburatani, H., Kodama, T., Ikai, A., Itakura, H., Akanuma, Y. and Takaku, F.:Analysis of apolipoproteins B-100 and B-48 by sodium dodecyl sulfate polyacrylamide gradient gel electrophoresis. J. Biochem., 94: (1983). 12) Meng, M. S., Gregg, R. E., Shaefer, E. J., Hoeg, J. M. and Brewer, H. B. Jr.:Presence of two forms of apolipoprotein B in patients with dyslipoproteinemia. J. Lipid Res., 24: (1983). 13) Fainaru, M., Mahley, R. W., Hamilton, R. L. and Innerarity, T. L.:Structural and metabolic hetero- cholesterol-fed dogs and humans with type III hyperlipoproteinemia. J. Lipid Res., 23: (1982). 14) Kane, J. P., Chen, G. C., Hamilton, D. A., Malloy, M. I. and Havel, R. J.:Remnants of lipoproteins of intestinal and hepatic origin in familial dysbetalipoproteinemia. Arteriosclerosis, 3:47-56(1983). 15) Cazzolato, G., Bon, G. B. and Avogaro, P.: Apoprotein B-48 is a constant finding in very low density lipoproteins of humans. Arteriosclerosis, 5:88-92(1985). 16) Nestel, P. J., Billington, T. and Fidge, N. H.: Slower removal of intestinal apolipoprotein B-48 than of apolipoprotein B-100 in severely hypertriglyceridemic subjects. Biochim. Biophys. Acta, 751: (1983). 17) Mime, R. W., Weech, P. K., Blanchette, L., Davignon, J., Alaupovic, P. and Marcel, Y. L.: 3) Marcel, Y. L., Hogue, M., Theolis, R. Jr. and Milne, R. W.:Mapping of Antigenic determinants of human apolipoprotein B using monoclonal antibodies against low density lipoproteins. J. Biol. Chem., 257: (1982). 4) Tikkanen, M. J., Cole, T. G., Hahm, K. S., Krul, E. S. and Schonfeld, G.:Expression of apolipoprotein B epitopes in very low density lipoprotein Isolation and characterization of apolipoprotein subfractions-studies with monoclonal antibodies-. B-48 and B-100 very low density lipoproteins from Arteriosclerosis, 4: (1984). type III hyperlipoproteinemic subjects. J. Clin. Invest., 73: (1984). 18) Terce, F., Mime, R. W., Weech, P. K., Davignon, J. and Marcel, Y. L.:Apolipoprotein B-48 and B- 100 very low density lipoproteins-comparison in 6) Mancini, G., Carbonara, A. O. and Heremans, dysbetalipoproteinemia (type III) and familial J. F.:Immunochemical quantitation of antigens by hypertriglyceridemia (type IV)-. Arteriosclerosis, single radial immunodiffusion. Int. J. Immunochem., 5: (1985). 2: (1965). 19) Van't Hooft, F. M., Hardman, D. A., Kane, J. P. 7) Sniderman, A., Teng, B. and Jerry, M.:Determination and Havel, R. J.:Apolipoprotein B (B-48) of rat of B protein of low density lipoprotein chylomicrons is not a precursor of the apolipopro- directly in plasma. J. Lipid Res., 16: tein of low density lipoprotein. Proc. Natl. Acad. (1975). Sci., 79: (1982). 20) Wu, A. L. and Windmueller, H. G.:Variant forms of plasma apolipoprotein B. J. Biol. Chem., 256: 9) Swaney, J. B. and Kuehl, K. S.:Separation of (1981). apolipoproteins by an acrylamide-gradient sodium
7 Summary Apolipoprotein B Subspecies of VLDL in Primary Hyperlipoproteinemia and Diabetes Mellitus Small molecular weight apo BS (B-48) originates from intestine and has different genetic regulation from large molecular weight apo BL (B-100). Apo BS will be a marker of exogenous lipoprotein. We have investigated apo BS concentration in VLDL of primary (especially type IIb and IV hyperlipoproteinemia which are considered to be caused by metabolic derangement of endogenous lipoprotein) and secondary hyperlipoproteinemia associated with diabetes mellitus. Concentrations of lipids in fasting plasma and lipoprotein fractions (VLDL, IDL, LDL and HDL) were determined enzymatically. Apo B subspecies in VLDL were analyzed by SDS-3-15% grandient polyacrylamide gel electrophoresis. Apo BS ratio (percentage of apo BS in total apo B) in normal control subjects proteinemia were significantly higher than control. Significant positive correlations were observed between apo BS ratio and plasma concentrations of triglyceride (TG), apo C-II and apo E. There was a significant negative correlation between apo BS ratio and plasma concentration of apo B. Type II diabetic patients showed a significantly higher apo significant correlations were not observed between apo BS ratio and plasma concentrations of TG, apo C-II and apo E. Most of apo BS ratios in the diabetics were plotted in the positions higher than the regression line in control and primary hyperlipoproteinemia. The data presented suggested the usefulness of apo BS ratio for understanding of metabolic derangement of exogenous lipoproteins. Key words:apolipoprtein B, VLDL, Hyperlipoproteinemia, Diabetes Mellitus
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