牛初乳中IGF-Ⅰ的分离及其降血糖作用研究
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摘要
本课题对牛初乳中IGF-I的分离及其对糖尿病大鼠的降糖作用进行了初步研究，主要研究结论如下：
牛初乳中IGF-I的含量随泌乳天数的推移急剧下降，泌乳前五
天，初乳中的IGF-I主要以游离形态存在，游离IGF-I含量占总IGF-I含量的比例由71.1%下降到50.64%，之后初乳中的IGF-I主要以结合形态存在，结合IGF-I含量占总IGF-I含量的比例由51.41%上升到60.1%。
采用酸－乙醇等前处理和Sephedex G-75凝胶过滤层析得到IGF-I分离物，前处理过程的平均收率为97.2%，层析过程的平均收率为58.84%,分离物中IGF-I的浓度平均达到2.137mg/g,较初乳提高约440倍。
糖尿病动物实验表明：IGF-I能降低糖尿病大鼠的血糖。IGF-I
的降糖效果呈现明显的剂量效应，口服15ugIGF-I/kg.bw组、10ug
IGF-I/kg.bw组和5ugIGF-I/kg.bw组大鼠的平均血糖值分别下降18.21%、14.96%和14.55%；腹腔注射的降糖效果明显好于口服，腹腔注射组大鼠的平均血糖值下降43.23%。IGF-I的降糖效应具有一定的持续性和稳定性。
大鼠血清激素测定结果表明：IGF-I能够升高血清胰岛素水平、下调生长激素水平以及增加血清游离IGF-I含量。
IGF-I治疗后糖尿病大鼠糖耐量曲线的下移提示IGF-I能够在一定程度上改善糖尿病大鼠的糖耐量，增加对葡萄糖的耐受性和利用性。
大鼠的肝组织、胰腺组织病理切片表明，IGF-I不仅能够有效地
促进血液中葡萄糖转化为肝糖元，而且能够促使剩余的胰岛β细胞发
生一定程度的再生。
根据本研究结果推断IGF-I的降糖机理为：通过促进肝糖元合成和促使剩余胰岛β细胞再生两方面来降低机体血糖水平。
The subject of this research is to separate insulin-like growth factor-Ⅰ(IGF-Ⅰ) from bovine colostrums and to observe its hypoglycemic effect in streptozotocin-induced diabetic rats.The results are as follow:
Concentrations of free and bound IGF-Ⅰwere measured in bovine colostrums from the
first milkings to 14 day postpartum.Rapid decrease was found during this period. IGF-Ⅰ
appeared mainly in the free form at 1-5 day colostrums and the ratio varied from 71.1% to 50.64%.After that,bound form dominated and the ratio varied from 51.41% to 60.1%.
Using the extraction of acid-ethanol and gel filtration chromatography methods, IGF-Ⅰwas isolated and purified from colostrums.The recovery of IGF-Ⅰwas 97.2% and 58.84%
respectively in two process.The content of IGF-Ⅰwas 2.137mg/g,440 times higher than colostrums.
The hypoglycemic effect of IGF-Ⅰon streptozotocin-induced diabetic rats was evaluated. The results were: ⑴the bovine colostrum IGF-Ⅰhad obvious hypoglycemic
effect in diabetic rats;⑵compared with basic value,plasma glucose value was decreased about 18.21%、14.96% and 14.55% in high,middle and low dose group by oral administration;⑶the glucose value of intraperitoneal group was about 43.23%,better than oral administration;⑷its hypoglycemic effect had persistence and stability.
Serum homone level of all rats were measured by radioimmunoassay. The results showed: IGF-Ⅰcould increase serum insulin and free IGF-Ⅰlevel significantly and decrease growth homone level.
The curve of glucose resistence descended obviously in administration group.It implied that IGF-Ⅰcould improve glucose resistence of diabetic rats to some degree.
The cell of liver and pancreas were observed by micrography.It showed that IGF-Ⅰcould accelerate the glycogenesis and promote the regeneration of residualβ-cell .
Based on research results,the hypoglycemic mechanism of IGF-Ⅰwas deduced.It is that ⑴By accelerating the glycogenesis ,IGF-Ⅰcan make good use of serum glucose.In this way,glucose utilization is increased.⑵IGF-Ⅰcan also have the residualβ-cell regenerated so that much more insulin is produced to exert hypoglycemic action.
牛初乳中IGF-I的含量随泌乳天数的推移急剧下降，泌乳前五
天，初乳中的IGF-I主要以游离形态存在，游离IGF-I含量占总IGF-I含量的比例由71.1%下降到50.64%，之后初乳中的IGF-I主要以结合形态存在，结合IGF-I含量占总IGF-I含量的比例由51.41%上升到60.1%。
采用酸－乙醇等前处理和Sephedex G-75凝胶过滤层析得到IGF-I分离物，前处理过程的平均收率为97.2%，层析过程的平均收率为58.84%,分离物中IGF-I的浓度平均达到2.137mg/g,较初乳提高约440倍。
糖尿病动物实验表明：IGF-I能降低糖尿病大鼠的血糖。IGF-I
的降糖效果呈现明显的剂量效应，口服15ugIGF-I/kg.bw组、10ug
IGF-I/kg.bw组和5ugIGF-I/kg.bw组大鼠的平均血糖值分别下降18.21%、14.96%和14.55%；腹腔注射的降糖效果明显好于口服，腹腔注射组大鼠的平均血糖值下降43.23%。IGF-I的降糖效应具有一定的持续性和稳定性。
大鼠血清激素测定结果表明：IGF-I能够升高血清胰岛素水平、下调生长激素水平以及增加血清游离IGF-I含量。
IGF-I治疗后糖尿病大鼠糖耐量曲线的下移提示IGF-I能够在一定程度上改善糖尿病大鼠的糖耐量，增加对葡萄糖的耐受性和利用性。
大鼠的肝组织、胰腺组织病理切片表明，IGF-I不仅能够有效地
促进血液中葡萄糖转化为肝糖元，而且能够促使剩余的胰岛β细胞发
生一定程度的再生。
根据本研究结果推断IGF-I的降糖机理为：通过促进肝糖元合成和促使剩余胰岛β细胞再生两方面来降低机体血糖水平。
The subject of this research is to separate insulin-like growth factor-Ⅰ(IGF-Ⅰ) from bovine colostrums and to observe its hypoglycemic effect in streptozotocin-induced diabetic rats.The results are as follow:
Concentrations of free and bound IGF-Ⅰwere measured in bovine colostrums from the
first milkings to 14 day postpartum.Rapid decrease was found during this period. IGF-Ⅰ
appeared mainly in the free form at 1-5 day colostrums and the ratio varied from 71.1% to 50.64%.After that,bound form dominated and the ratio varied from 51.41% to 60.1%.
Using the extraction of acid-ethanol and gel filtration chromatography methods, IGF-Ⅰwas isolated and purified from colostrums.The recovery of IGF-Ⅰwas 97.2% and 58.84%
respectively in two process.The content of IGF-Ⅰwas 2.137mg/g,440 times higher than colostrums.
The hypoglycemic effect of IGF-Ⅰon streptozotocin-induced diabetic rats was evaluated. The results were: ⑴the bovine colostrum IGF-Ⅰhad obvious hypoglycemic
effect in diabetic rats;⑵compared with basic value,plasma glucose value was decreased about 18.21%、14.96% and 14.55% in high,middle and low dose group by oral administration;⑶the glucose value of intraperitoneal group was about 43.23%,better than oral administration;⑷its hypoglycemic effect had persistence and stability.
Serum homone level of all rats were measured by radioimmunoassay. The results showed: IGF-Ⅰcould increase serum insulin and free IGF-Ⅰlevel significantly and decrease growth homone level.
The curve of glucose resistence descended obviously in administration group.It implied that IGF-Ⅰcould improve glucose resistence of diabetic rats to some degree.
The cell of liver and pancreas were observed by micrography.It showed that IGF-Ⅰcould accelerate the glycogenesis and promote the regeneration of residualβ-cell .
Based on research results,the hypoglycemic mechanism of IGF-Ⅰwas deduced.It is that ⑴By accelerating the glycogenesis ,IGF-Ⅰcan make good use of serum glucose.In this way,glucose utilization is increased.⑵IGF-Ⅰcan also have the residualβ-cell regenerated so that much more insulin is produced to exert hypoglycemic action.
引文
1 陆东林，张丹凤主编.奶牛初乳及其保健功能研究.乌鲁木齐：新疆人民出版社，
2001.8
2 Grosvenor,C.E.,Picciano,M.F.and Baumrucker,C.R.(1992) Endocr.Rev.14,710-728
3 Schams,D.(1994) Endocr.Regul.28,3-8
4 Vega,J.R.,Gibson,C.A.,Skaar,T.C.,Hadsell,D.L.and Baumrucker,C.R.(1991) J.Amin.Sci.
69,2538-2547
5 向红丁等.1996年全国糖尿病流行病学特点基线调查报告.中国糖尿病杂志，1998,3:131-133
6 www.bioon.com/药物研究/糖尿病流行病学调查报告
7 何戎华等. 糖尿病现代诊疗.南京：江苏科学技术出版社，2000.9
8 池芝盛.拜糖平的临床应用.中国糖尿病杂志，1995,3:165
9 Pickup Jand Williams G.Textbook of diabetes.2 edition by Blackwell of Science Ltd, 1997
10 Salmon,W.D.,Daughaday,W.H.[J] J Lab Clin Med,1957,49:825-836
11 Daughaday,W.H.,Hall,K.,Salmon,W.D.,et al. [J] Clin Endocrinal Metab,1987,65:1075
-1076
12 段宇等.胰岛素样生长因子Ⅰ研究进展.国外医学内分泌学分册,2001,11(6):305-
306
13 Hauser,S.D.,Mcgrath,M.F.,Collier,R.J.& Krivi,G.G. Cloning and in vivo expression of
bovine growth hormone receptor mRNA.J.Molecular and cellular Endocrinology.
1990,72:187-200
14 Zumstein,P.P.& Humbel,R.E.(1985) Methods Enzymol.109,part 1,782-798
15 Blum,W.F.,Ranke,M.B.&Bierich,J.R.(1986) Acta Endocrinol,111,271-284
16 Francis,G.L.,Read,L.C.,Ballard,F.J.,et al.(1986)Biochem.J.223,207-213
17魏辉俊,黄秉仁.类胰岛素生长因子Ⅰ的克隆、表达、分离纯化与活性检测.中国医
学科学院学报,1997,19(5):342-345
18 吴晓岚李明等.人胰岛素生长因子Ⅰ基因的克隆及其表达.中国生化药物杂志, 2002,23(1):1-3
19 Svoboda,M.E.,Van,J.J.,Klapper,D.G.,et al.(1980) Biochem.J.19,790-797
20 Clemmons,D.R.,Underwood,L.E.[J].Annu Rev Nutr,1991,11:393-412
21 King,G.L.,Kahn,C.R.,Samuels,B.,Danho,W.,et al.(1982) J.Bio.Chem.257,10869-10873
22 Tseng,L.Y.,Schwartz,G.P.,Sheik,M.,Chen,Z.Z.,Joshi,S.,et al.(1986)Fed.Am.Soc.Exp.
Bio.45,1714
23 De Vroede,M.A.,Rechler,M.M.,Nissley,S.P.,Ogawa,H.,Burke,G.T.,(1986) Diabetes.35,
355-361
24 Blundell,T.L.,Bedarkar,S.& Humbel,R.E.(1983) Fed.Proc.Fed.Am.Soc.Exp.Bio.42,
2592-2597
25 Honegger,A. & Humbel,R.E.(1986) J.Biol.Chem.261,569-575
26 Bell,G.I.,Stempein,M.M.,Fong,N.M.&Rall,L.B.(1986)Nucleic Acids Res.14,7873-7882
27王利华，周丹.类胰岛素生长因子生理及在畜牧业中的应用.畜禽业，2001,10:14-15
28 Anthony,S.et al.(1990) J.Biochem Biophy Res Comm.166,892-897
29 魏辉俊.类胰岛素生长因子结合蛋白.国外医学内分泌学分册,1994,4:175-177
30 Campbell,P.G.and Baumrucker,C.R. Insulin-like growth factorⅠand its
association with binding proteins in bovine milk. 1989 .J.Endocrinol. 120, 21-29
31 Skaar,T.C.,Vega,S.N.and Baumrucker,C.R. Changes in insulin-like growth factor-
binding proteins in bovine mammary secretions associated with pregnancy and parturit-
ion.J.Endocrinol.1991,131,127-133
32 Eriksson,U.G.,Froesch,E.R.and Zapf. Insulin-like growth factorⅠandⅡ
and IGF-binding proteins in Human colostrums/Transitory milk During the
First week postpartum:Comparison with neonatal and maternal serum. J.
Biochem Biophy Res Comm. 1993,196,1:267-273
33 Ralf Einspanier and Dieter Schams. Changes in concentrations insulin-like growth
hormone in bovine mammary gland secretion ante and post partum.J.DairResearch.
1991,58,171-178
34 Skaar,T.C., Baumrucker,C.R.,Deaver,D.R.,and Blum,J.W.Diet Effect and Ontogeny of Alterations of Circulating insulin-like growth factor Binding Proteins in Newborn
Dairy Calves.J. Anim. Sci.1994,72:421-427
35 路聪哲.IGF-I与IGF-I-BP1在GRAVES病糖代谢中的应用.国外医学内分泌学分册,2001,4:212-213
36 曹劲松主编. 初乳功能性食品. 北京：中国轻工业出版社，2000.3:91-97
37 Ballard,F.J.,John,C.,Wallace,Geoffrey,L.,Francis,Leanna,C.,Read,Frank,M.,Tomas.Des
(1-3) IGF-Ⅰ:a Truncated Form of insulin-like growth factor1.J.Biochem.Cell Bio1996
10:1085-1087
38 Francis,G.L.,Upton,F.M.,Ballard,F.J.,Mcneil,K.A.and Wallace,J.C.
Insulin-likegrowth factor1 and 2 in bovine colostrums:sequences and
biological activities compared with those of a potent truncated form. J.Biochem.1988,
251:95-103
39 Gilmour,R.S.The implications of Insulin-like growth factor mRNA
heterogeneity.J.Endocrinol. 1994,140:1-3
40 Yamamoto,H.and Murphy,L.J.Generation of des(1-3) insulin-like growth factor1 in serum by an acid protease.Endocrinology.1994,135:2432-2439
41 Bagley,C.J.,May,B.L.,Szabo,L.,Mcnamara,P.J.,Ross,M.,Francis,G.L.,Ballard,F.J and
Wallace,J.C.A key functional role for the insulin-like growth factor 1 N-terminal pentapeptide. J.Biochem.1989,259:665-671.
42 Ogasawara,M.,Karey,K.P.,Marquardt,H.and Sirbasku,D.A.Identification and
purification of truncated insulin-like growth factor 1 from porcine uterus.
Evidence for high biological activity.Biochemistry.1989,28:2710-2721.
43 Sara,V.R.,Carlsson-Skwirut,C.,Andersson,C.,Hall,E.,and Jornvall.Characterizations of
somatomedins from human fetal brain:identification of a Variant form of insulin-like growth factor 1.Proc.Natl Acad.Sci.USA.1986,83:4904-4907.
44 Ballard,F.J.,Knowles,S.E.,Waiton,P.E.,Edson,K.,Owens,P.C.,Mohler,M.A.and Ferraiolo
B.L.Plasma clearance and tissue distribution of labeled insulin-Like growth factor Ⅰ,
IGF-Ⅱ,and des(1-3)IGF-Ⅰin rats. J.Endocrinol.1991,128:197-204
45 Walton,P.E.,Dunshea,F.R.,and Ballard,F.J.Invivo actions of IGF analogues with poor affinities for IGFBPS:metabolic and growth effects in pigs of differentages and GH
responsiveness. Progr Growth Factor Res.1995,6:385-395
46 张克英，陈代文.类胰岛素生长因子生理及其营养调控.中国畜牧杂志,2002,6:42
-44
47 Froesch,E.R.,Schmid,C.,Schwander,J.,et al.Function of Insulin-like growth
factor.J.Annu Rev Nutr.1991,11:393-412
48 Francis,G.L.,Upton,F.M.,Ballard,F.J.,Mcneil,K.A.and Wallace,J.C.Insulin-like growth
factors 1 and 2 in bovine colostrums:sequences and biological activities compared
with those of a potent truncated form. J.Biochem.1988,251:95-103
49 Young,G.P.,Taranto,T.M.,Jonas,H.A.,Cox,A.J.,Hogg,A.andWerther,G.A. Insulin-like
growth factor and the developing and mature rat small intestine:receptor and biogical
vactions.J.Digestion.1990,46:240-252
50 Zhang,W.,Frankel,W.L.,Adamson,W.T.,Roth,J.A.,Msntell,M.P.,Bain,A.,
Ziegler,T.R.,Smith,R.J.and Rombeau,J.L. Insulin-like growth factors 1
improves mucosal Structure and function in transplanted rat small intestine.J. Trans-
plantation 1995,59(5):755-761
51 Zumkeller,W.Relationship between insulin-Like growth factorⅠand IGF-Ⅱand IGF-binding proteins in milk and the gastrointestinal tract:growth
and Development of the gut. J.Pediatr.Gastroenterol.Nutr.1992,15:357-369
52 Karin Fholenhag,Vibeke Arrhenius-Nyberg,Ingrid Sjogren and Kjell Malmlof.
Effect of insulin-Like growth factorⅠ(IGF-Ⅰ)on the small intestine: A
Comparison between oral and subcutaneous administration in the weaned rat.
J.Growth factors.1997,14:81-88
53 钱洪松，张光霁，朱诚等.IGF-Ⅰ与中枢神经系统的研究进展.国外医学生理、病理科学与临床分册，1996,4:272-274
54 Kenneth,K.A.,Drogo,J.D.,Selinfreund,R.H.et al.Arch Surg.1994,129:66-68
55 Malven,P.V.,Head,H.H.,Collier,R.J.&Buonomo,F.C.Periparturient changes in
secretion and mammary uptake of insulin and in concentrations of insulin and insulin-Like growth factors in milk of dairy cows. J. Dairy Science.
1987,70:2254-2265
56 Robert,J.Collier,Miller,M.A.,Hildebrandt,J.R.,Torkelson,A.R.,White,K.S.,Madsen,K.S.
Vicini,J.L.,Eppard,P.J.and Lanza,G.M.Factors Affecting Insulin-Like Growth Factor-ⅠConcentration in Bovine Milk. J.Dairy Science.1991,74:2905-2911
57 Hadsell,D.L.,Baumrucker and Kensinger.Effects of elevated blood insulin-
Like growth factorⅠ(IGF-Ⅰ)concentration upon IGF-Ⅰin bovine mammary secretions during the colostrums phase.J.Endocrinology.1993,137:223-230
58 Davis,S.R.,Gluckman,P.,Hart,I.C.&Henderson,H.V.Effectsof injecting
growth hormone or thyroxine on milk production and blood plasma
concentration of insulin-Like growth factorⅠand IGF-Ⅱ in dairy cows.J.
Endocrinology.1987,114:17-24
59 Prosser,C.G.,Fleet,I.R.& Corps,A.N.Increased secretion of insulin-Like growth factorⅠinto milk of cows treated with recombinantly derived bovine growth hormone.J.
Dairy Research.1989,56:17-26
60 Glimm,D.R.,Baracos,V.E.& Kennelly,J.J.Effect of bovine somatotropin on the
distribution of immunoreactive insulin-Like growth factorⅠin lactating
bovine mammary tissue.J.Dairy Science.1988,71:2923-2935
61 Murphy,L.J.,Bell,G.I.& Friesen,H.G.Tissue distribution of insulin-Like growth and IGF-Ⅱmessenger ribonucleic acid in the adult rat.J.Endocrinology.1987,120:1279-
1282
62 Hauser,S.D.,Mcgrath,M.F.,Collier,R.J.& Krivi,G.G. Cloning and in vivo
expression of bovine growth hormone receptor mRNA.J.Molecular and Cellular
Endocrinology.1990,72:187-200
63 Campbell,P.G.,Skaar,T.C.,Vega,J.R.& Baumrucker,C.R.Secretion of insulin-
Like factorⅠ(IGF-Ⅰ)and IGF-binding proteins from bovine mammary tissue
in vivo.J.Endocrinology.1991,128:219-228
64 Sasaki,T.C.,Davis,C.L.,& Larson,B.L.Production and turnover of IgG1 and IgG2immunoglobulins in the bovine around parturition.J.Dairy Science.
1976,59:2046-2055
65 Brown,K.D.,Blakeley,D.M.,Fleet,I.R.,Hamon,M.& Heap,R.B.Kinetics of
transfer125I-labelled epidermal growth factor from blood into mammary
secretions of goats.J. Endocrinology.1986,109:325-332
66 Heap,R.B.,Hamon,M.& Fleet,I.R.Transport of oestrone sulphate by the mammary gland
in the goat.J. Endocrinology.1984,101:221-230
67 Vega,J.R.,Gibson,C.A.,Skaar,T.C.,Hadsell,D.L.& Baumrucker,C.R. Insulin-
Like factorⅠ(IGF-Ⅰ)and IGF-Ⅱand IGF-binding proteins in serum and
Mammary secretions during the dry period and early lactation in dairy cows.J.Animal Science.1991,69:2538-2547
68 Prosser,C.G.,Fleet,I.R.,Davis,A.J.and Heap,R.B. Mechanism of secretion of
plasma insulin-Like growth factorⅠinto milk of lactating goats.J.
Endocrinology.1991,131:459-466
69 Read,L.C.,Gale,S.M.&George-Nascimento,C.Absorption of immunologically-
intact proteins by the auto-perfused lamb intestine:comparative uptake of
epidermal growth factor and immunoglobulin. American Journal of Physiology.
1988,12
70 Young,G.P.,Taranto,T.M.,Jones,H.A.,Cox,A.J.,Hogg,A.and Werther,G.A.
Insulin-Like growth factor and the developing and mature rat small intestine:Receptors And biological actions. Digestion.1990,46:240-252
71 Xu,R.J.,Mellor,D.J.,Birtles,B.H.and Gluckmam,P.D. Effect of oral IGF-Ⅰor IGF-Ⅱon digestive organ growth in newborn piglets. J.biol Neonate.1994,66:280-287
72 Gallo-Payet,N.,Pothier,P.& Hugon,J.S. Ontogeny of EGF receptors during the postnatal development of mouse small intestine.Journal of Pediatric Gastroenterology and Nutrition.1987,6:114-120
73 Olanrewaju,H.,Patel,L.and Scidel,E.R.Trophic action of local intraileal
infusion of insulin-Like growth factorⅠ:polyaminc dependence. Am.J.
Physiol.1992,263:E282-E286
74 Pierce,A.E.,Risdall,P.C.& Shaw,B. Adsorption of orally administered
insulin by the newly born calf.Journal of Physiology.1964,171:203-215
75 胡绍文，郭瑞林主编.实用糖尿病学.北京：人民军医出版社,1998
76 朱忠勇主编.实用医学检验学. 北京：人民军医出版社,1992
77 蒋国彦主编.实用糖尿病学. 北京：人民卫生出版社，1997
78 何戎华主编.糖尿病200问.南京：江苏科学技术出版社,1996
79 陈琰，胡晋红，朱全刚.糖尿病发病机理的讨论.上海药学服务，2000，1：12-16
80 www.37c.com.cn/tangniaobing/(37℃医学网)
81李冬梅，雷琪智.糖尿病必读手册.呼和浩特：内蒙古人民出版社，2003，1
82 www.chinadiabetes.net(中国糖尿病信息网)
83 何戎华.糖尿病几种亚型的合理用药及临床处理与评价.实用糖尿病杂志,1998,6:11-13
84 石根勇，吕中明，陈新霞，凌宝银，张惠菊.降糖茶调节血糖作用的研究.中国生化药物杂志,2001,5:249-250
85 陈海霞，谢笔钧.茶多糖对小鼠实验性糖尿病的防治作用.营养学报,2002,1:85-86
86 郜文.保健南瓜粉生产工艺研究.食品工业科技,2001,22(5):63-64
87 左耀明,叶士伶,万小兰,樊惠民等.南瓜多糖的提取分析和降血糖试验研究.食品科学,2001,22(12):56-58
88 王亚伟，李春亚.膳食纤维和低聚糖对糖尿病人血糖血脂的影响.郑州工程学院学报,2002,23(1):85-88
89王亚伟,王燕等.膳食纤维对糖尿病人血糖的影响.食品工业技,2001,22(5):25-27
90 Elias,A.N.,Grossman,M.K.,Valenta,L.J.,et al.Use of the artificial beta cell in the assessment of peripheral insulin sensitivity:effect of chromium supple-mentation in
diabetic patients. J.Gen Pharmacol.1984,15:535
91 赵晓华，李兴，宋征，杨燕等.加铬复合纤维对糖尿病大鼠治疗作用观察.营养学报,2000,4:312-316
92 姜桂荣等.富铬酵母对糖尿病人糖耐量的影响.营养学报,1988,10(3):253
93 Osamu Ezaki.The insulin-like effects of selenate in rat adipocytes.J.Biol Chem.1990,
265:1124
94 丁虹，彭仁琇，陈金和，黎七雄，汪晖，孔锐.硒、大蒜素对糖尿病小鼠降糖作用的研究.营养学报,1997,4:384-386
95 Ballard,F.J.,Tomas,F.M.,Read,L.C.,Knowles,S.E.,Owens,j.c.and Francis,
G.L.Effect of IGF-Ⅰand IGF analogs on growth during catabolic states
in rat.In Modern Concepts of Insulin-like Growth Factors.
(Edited by Spencer,E.M.)Elsevier,New York.1991,617-627
96 Read,l.c.,Lemmey,A.B.,Howarth,G.S.,Martin,A.A.,Tomas,F.M.and Ballard,
F.J.The gastrointestinal tract is one of the most responsive target
tissues forIGF-Ⅰand its potent analogs. In Modern Concepts of Insulin-like
Growth Factors.(Edited by Spencer,E.M.) Elsevier,New York.1991,225-234
97 叶红英，俞茂华，游利，颜贻谦.外源性牛初乳短链IGF-Ⅰ治疗改善糖尿病大鼠GH/ IGF-Ⅰ轴的紊乱.中国糖尿病杂志.2002,1:15-16
98 孙侃，俞茂华，史虹莉，杨秀芳，方京冲等.牛初乳短链IGF-Ⅰ对STZ糖尿病大鼠物质代谢及肾脏的影响.中华内分泌代谢杂志.1997,3:162-166
99 俞茂华,朱禧星,史虹莉,何德华等.牛初乳制剂治疗型糖尿病的临床初步观察.上海医科大学学报,1992,19(1):41-44
100 Etherton,T.D.,Wiggins,J.P.,Evock,C.M.,Chung,C.S.,Rebhun,J.F.,Walton,
P.E.& Steele,N.C. Stimulation of pig growth performance by porcine
growth hormone:Determination of the dose-response relationship.J.
Animal Science.1987,64:433-443
101 张龙翔,张庭芳,李令媛主编.生化实验方法和技术.北京:高等教育出版社,1997,136-137
102 徐叔云,卡如濂,陈修主编.药理实验方法学（第二版）.北京:人民卫生出版社,1995,1277-1278
103 H.G沃格尔[德],W.H.沃格尔[美]编著.杜冠华,李学军,张永祥等译.药理学实验指南－新药发现和药理学评价.北京:科学出版社,2001,2:700-701
104 Rao,R.K. Biologically active peptides in the gastrointestinal lumen.Life
Sci.1991,48:1685-1704
2001.8
2 Grosvenor,C.E.,Picciano,M.F.and Baumrucker,C.R.(1992) Endocr.Rev.14,710-728
3 Schams,D.(1994) Endocr.Regul.28,3-8
4 Vega,J.R.,Gibson,C.A.,Skaar,T.C.,Hadsell,D.L.and Baumrucker,C.R.(1991) J.Amin.Sci.
69,2538-2547
5 向红丁等.1996年全国糖尿病流行病学特点基线调查报告.中国糖尿病杂志，1998,3:131-133
6 www.bioon.com/药物研究/糖尿病流行病学调查报告
7 何戎华等. 糖尿病现代诊疗.南京：江苏科学技术出版社，2000.9
8 池芝盛.拜糖平的临床应用.中国糖尿病杂志，1995,3:165
9 Pickup Jand Williams G.Textbook of diabetes.2 edition by Blackwell of Science Ltd, 1997
10 Salmon,W.D.,Daughaday,W.H.[J] J Lab Clin Med,1957,49:825-836
11 Daughaday,W.H.,Hall,K.,Salmon,W.D.,et al. [J] Clin Endocrinal Metab,1987,65:1075
-1076
12 段宇等.胰岛素样生长因子Ⅰ研究进展.国外医学内分泌学分册,2001,11(6):305-
306
13 Hauser,S.D.,Mcgrath,M.F.,Collier,R.J.& Krivi,G.G. Cloning and in vivo expression of
bovine growth hormone receptor mRNA.J.Molecular and cellular Endocrinology.
1990,72:187-200
14 Zumstein,P.P.& Humbel,R.E.(1985) Methods Enzymol.109,part 1,782-798
15 Blum,W.F.,Ranke,M.B.&Bierich,J.R.(1986) Acta Endocrinol,111,271-284
16 Francis,G.L.,Read,L.C.,Ballard,F.J.,et al.(1986)Biochem.J.223,207-213
17魏辉俊,黄秉仁.类胰岛素生长因子Ⅰ的克隆、表达、分离纯化与活性检测.中国医
学科学院学报,1997,19(5):342-345
18 吴晓岚李明等.人胰岛素生长因子Ⅰ基因的克隆及其表达.中国生化药物杂志, 2002,23(1):1-3
19 Svoboda,M.E.,Van,J.J.,Klapper,D.G.,et al.(1980) Biochem.J.19,790-797
20 Clemmons,D.R.,Underwood,L.E.[J].Annu Rev Nutr,1991,11:393-412
21 King,G.L.,Kahn,C.R.,Samuels,B.,Danho,W.,et al.(1982) J.Bio.Chem.257,10869-10873
22 Tseng,L.Y.,Schwartz,G.P.,Sheik,M.,Chen,Z.Z.,Joshi,S.,et al.(1986)Fed.Am.Soc.Exp.
Bio.45,1714
23 De Vroede,M.A.,Rechler,M.M.,Nissley,S.P.,Ogawa,H.,Burke,G.T.,(1986) Diabetes.35,
355-361
24 Blundell,T.L.,Bedarkar,S.& Humbel,R.E.(1983) Fed.Proc.Fed.Am.Soc.Exp.Bio.42,
2592-2597
25 Honegger,A. & Humbel,R.E.(1986) J.Biol.Chem.261,569-575
26 Bell,G.I.,Stempein,M.M.,Fong,N.M.&Rall,L.B.(1986)Nucleic Acids Res.14,7873-7882
27王利华，周丹.类胰岛素生长因子生理及在畜牧业中的应用.畜禽业，2001,10:14-15
28 Anthony,S.et al.(1990) J.Biochem Biophy Res Comm.166,892-897
29 魏辉俊.类胰岛素生长因子结合蛋白.国外医学内分泌学分册,1994,4:175-177
30 Campbell,P.G.and Baumrucker,C.R. Insulin-like growth factorⅠand its
association with binding proteins in bovine milk. 1989 .J.Endocrinol. 120, 21-29
31 Skaar,T.C.,Vega,S.N.and Baumrucker,C.R. Changes in insulin-like growth factor-
binding proteins in bovine mammary secretions associated with pregnancy and parturit-
ion.J.Endocrinol.1991,131,127-133
32 Eriksson,U.G.,Froesch,E.R.and Zapf. Insulin-like growth factorⅠandⅡ
and IGF-binding proteins in Human colostrums/Transitory milk During the
First week postpartum:Comparison with neonatal and maternal serum. J.
Biochem Biophy Res Comm. 1993,196,1:267-273
33 Ralf Einspanier and Dieter Schams. Changes in concentrations insulin-like growth
hormone in bovine mammary gland secretion ante and post partum.J.DairResearch.
1991,58,171-178
34 Skaar,T.C., Baumrucker,C.R.,Deaver,D.R.,and Blum,J.W.Diet Effect and Ontogeny of Alterations of Circulating insulin-like growth factor Binding Proteins in Newborn
Dairy Calves.J. Anim. Sci.1994,72:421-427
35 路聪哲.IGF-I与IGF-I-BP1在GRAVES病糖代谢中的应用.国外医学内分泌学分册,2001,4:212-213
36 曹劲松主编. 初乳功能性食品. 北京：中国轻工业出版社，2000.3:91-97
37 Ballard,F.J.,John,C.,Wallace,Geoffrey,L.,Francis,Leanna,C.,Read,Frank,M.,Tomas.Des
(1-3) IGF-Ⅰ:a Truncated Form of insulin-like growth factor1.J.Biochem.Cell Bio1996
10:1085-1087
38 Francis,G.L.,Upton,F.M.,Ballard,F.J.,Mcneil,K.A.and Wallace,J.C.
Insulin-likegrowth factor1 and 2 in bovine colostrums:sequences and
biological activities compared with those of a potent truncated form. J.Biochem.1988,
251:95-103
39 Gilmour,R.S.The implications of Insulin-like growth factor mRNA
heterogeneity.J.Endocrinol. 1994,140:1-3
40 Yamamoto,H.and Murphy,L.J.Generation of des(1-3) insulin-like growth factor1 in serum by an acid protease.Endocrinology.1994,135:2432-2439
41 Bagley,C.J.,May,B.L.,Szabo,L.,Mcnamara,P.J.,Ross,M.,Francis,G.L.,Ballard,F.J and
Wallace,J.C.A key functional role for the insulin-like growth factor 1 N-terminal pentapeptide. J.Biochem.1989,259:665-671.
42 Ogasawara,M.,Karey,K.P.,Marquardt,H.and Sirbasku,D.A.Identification and
purification of truncated insulin-like growth factor 1 from porcine uterus.
Evidence for high biological activity.Biochemistry.1989,28:2710-2721.
43 Sara,V.R.,Carlsson-Skwirut,C.,Andersson,C.,Hall,E.,and Jornvall.Characterizations of
somatomedins from human fetal brain:identification of a Variant form of insulin-like growth factor 1.Proc.Natl Acad.Sci.USA.1986,83:4904-4907.
44 Ballard,F.J.,Knowles,S.E.,Waiton,P.E.,Edson,K.,Owens,P.C.,Mohler,M.A.and Ferraiolo
B.L.Plasma clearance and tissue distribution of labeled insulin-Like growth factor Ⅰ,
IGF-Ⅱ,and des(1-3)IGF-Ⅰin rats. J.Endocrinol.1991,128:197-204
45 Walton,P.E.,Dunshea,F.R.,and Ballard,F.J.Invivo actions of IGF analogues with poor affinities for IGFBPS:metabolic and growth effects in pigs of differentages and GH
responsiveness. Progr Growth Factor Res.1995,6:385-395
46 张克英，陈代文.类胰岛素生长因子生理及其营养调控.中国畜牧杂志,2002,6:42
-44
47 Froesch,E.R.,Schmid,C.,Schwander,J.,et al.Function of Insulin-like growth
factor.J.Annu Rev Nutr.1991,11:393-412
48 Francis,G.L.,Upton,F.M.,Ballard,F.J.,Mcneil,K.A.and Wallace,J.C.Insulin-like growth
factors 1 and 2 in bovine colostrums:sequences and biological activities compared
with those of a potent truncated form. J.Biochem.1988,251:95-103
49 Young,G.P.,Taranto,T.M.,Jonas,H.A.,Cox,A.J.,Hogg,A.andWerther,G.A. Insulin-like
growth factor and the developing and mature rat small intestine:receptor and biogical
vactions.J.Digestion.1990,46:240-252
50 Zhang,W.,Frankel,W.L.,Adamson,W.T.,Roth,J.A.,Msntell,M.P.,Bain,A.,
Ziegler,T.R.,Smith,R.J.and Rombeau,J.L. Insulin-like growth factors 1
improves mucosal Structure and function in transplanted rat small intestine.J. Trans-
plantation 1995,59(5):755-761
51 Zumkeller,W.Relationship between insulin-Like growth factorⅠand IGF-Ⅱand IGF-binding proteins in milk and the gastrointestinal tract:growth
and Development of the gut. J.Pediatr.Gastroenterol.Nutr.1992,15:357-369
52 Karin Fholenhag,Vibeke Arrhenius-Nyberg,Ingrid Sjogren and Kjell Malmlof.
Effect of insulin-Like growth factorⅠ(IGF-Ⅰ)on the small intestine: A
Comparison between oral and subcutaneous administration in the weaned rat.
J.Growth factors.1997,14:81-88
53 钱洪松，张光霁，朱诚等.IGF-Ⅰ与中枢神经系统的研究进展.国外医学生理、病理科学与临床分册，1996,4:272-274
54 Kenneth,K.A.,Drogo,J.D.,Selinfreund,R.H.et al.Arch Surg.1994,129:66-68
55 Malven,P.V.,Head,H.H.,Collier,R.J.&Buonomo,F.C.Periparturient changes in
secretion and mammary uptake of insulin and in concentrations of insulin and insulin-Like growth factors in milk of dairy cows. J. Dairy Science.
1987,70:2254-2265
56 Robert,J.Collier,Miller,M.A.,Hildebrandt,J.R.,Torkelson,A.R.,White,K.S.,Madsen,K.S.
Vicini,J.L.,Eppard,P.J.and Lanza,G.M.Factors Affecting Insulin-Like Growth Factor-ⅠConcentration in Bovine Milk. J.Dairy Science.1991,74:2905-2911
57 Hadsell,D.L.,Baumrucker and Kensinger.Effects of elevated blood insulin-
Like growth factorⅠ(IGF-Ⅰ)concentration upon IGF-Ⅰin bovine mammary secretions during the colostrums phase.J.Endocrinology.1993,137:223-230
58 Davis,S.R.,Gluckman,P.,Hart,I.C.&Henderson,H.V.Effectsof injecting
growth hormone or thyroxine on milk production and blood plasma
concentration of insulin-Like growth factorⅠand IGF-Ⅱ in dairy cows.J.
Endocrinology.1987,114:17-24
59 Prosser,C.G.,Fleet,I.R.& Corps,A.N.Increased secretion of insulin-Like growth factorⅠinto milk of cows treated with recombinantly derived bovine growth hormone.J.
Dairy Research.1989,56:17-26
60 Glimm,D.R.,Baracos,V.E.& Kennelly,J.J.Effect of bovine somatotropin on the
distribution of immunoreactive insulin-Like growth factorⅠin lactating
bovine mammary tissue.J.Dairy Science.1988,71:2923-2935
61 Murphy,L.J.,Bell,G.I.& Friesen,H.G.Tissue distribution of insulin-Like growth and IGF-Ⅱmessenger ribonucleic acid in the adult rat.J.Endocrinology.1987,120:1279-
1282
62 Hauser,S.D.,Mcgrath,M.F.,Collier,R.J.& Krivi,G.G. Cloning and in vivo
expression of bovine growth hormone receptor mRNA.J.Molecular and Cellular
Endocrinology.1990,72:187-200
63 Campbell,P.G.,Skaar,T.C.,Vega,J.R.& Baumrucker,C.R.Secretion of insulin-
Like factorⅠ(IGF-Ⅰ)and IGF-binding proteins from bovine mammary tissue
in vivo.J.Endocrinology.1991,128:219-228
64 Sasaki,T.C.,Davis,C.L.,& Larson,B.L.Production and turnover of IgG1 and IgG2immunoglobulins in the bovine around parturition.J.Dairy Science.
1976,59:2046-2055
65 Brown,K.D.,Blakeley,D.M.,Fleet,I.R.,Hamon,M.& Heap,R.B.Kinetics of
transfer125I-labelled epidermal growth factor from blood into mammary
secretions of goats.J. Endocrinology.1986,109:325-332
66 Heap,R.B.,Hamon,M.& Fleet,I.R.Transport of oestrone sulphate by the mammary gland
in the goat.J. Endocrinology.1984,101:221-230
67 Vega,J.R.,Gibson,C.A.,Skaar,T.C.,Hadsell,D.L.& Baumrucker,C.R. Insulin-
Like factorⅠ(IGF-Ⅰ)and IGF-Ⅱand IGF-binding proteins in serum and
Mammary secretions during the dry period and early lactation in dairy cows.J.Animal Science.1991,69:2538-2547
68 Prosser,C.G.,Fleet,I.R.,Davis,A.J.and Heap,R.B. Mechanism of secretion of
plasma insulin-Like growth factorⅠinto milk of lactating goats.J.
Endocrinology.1991,131:459-466
69 Read,L.C.,Gale,S.M.&George-Nascimento,C.Absorption of immunologically-
intact proteins by the auto-perfused lamb intestine:comparative uptake of
epidermal growth factor and immunoglobulin. American Journal of Physiology.
1988,12
70 Young,G.P.,Taranto,T.M.,Jones,H.A.,Cox,A.J.,Hogg,A.and Werther,G.A.
Insulin-Like growth factor and the developing and mature rat small intestine:Receptors And biological actions. Digestion.1990,46:240-252
71 Xu,R.J.,Mellor,D.J.,Birtles,B.H.and Gluckmam,P.D. Effect of oral IGF-Ⅰor IGF-Ⅱon digestive organ growth in newborn piglets. J.biol Neonate.1994,66:280-287
72 Gallo-Payet,N.,Pothier,P.& Hugon,J.S. Ontogeny of EGF receptors during the postnatal development of mouse small intestine.Journal of Pediatric Gastroenterology and Nutrition.1987,6:114-120
73 Olanrewaju,H.,Patel,L.and Scidel,E.R.Trophic action of local intraileal
infusion of insulin-Like growth factorⅠ:polyaminc dependence. Am.J.
Physiol.1992,263:E282-E286
74 Pierce,A.E.,Risdall,P.C.& Shaw,B. Adsorption of orally administered
insulin by the newly born calf.Journal of Physiology.1964,171:203-215
75 胡绍文，郭瑞林主编.实用糖尿病学.北京：人民军医出版社,1998
76 朱忠勇主编.实用医学检验学. 北京：人民军医出版社,1992
77 蒋国彦主编.实用糖尿病学. 北京：人民卫生出版社，1997
78 何戎华主编.糖尿病200问.南京：江苏科学技术出版社,1996
79 陈琰，胡晋红，朱全刚.糖尿病发病机理的讨论.上海药学服务，2000，1：12-16
80 www.37c.com.cn/tangniaobing/(37℃医学网)
81李冬梅，雷琪智.糖尿病必读手册.呼和浩特：内蒙古人民出版社，2003，1
82 www.chinadiabetes.net(中国糖尿病信息网)
83 何戎华.糖尿病几种亚型的合理用药及临床处理与评价.实用糖尿病杂志,1998,6:11-13
84 石根勇，吕中明，陈新霞，凌宝银，张惠菊.降糖茶调节血糖作用的研究.中国生化药物杂志,2001,5:249-250
85 陈海霞，谢笔钧.茶多糖对小鼠实验性糖尿病的防治作用.营养学报,2002,1:85-86
86 郜文.保健南瓜粉生产工艺研究.食品工业科技,2001,22(5):63-64
87 左耀明,叶士伶,万小兰,樊惠民等.南瓜多糖的提取分析和降血糖试验研究.食品科学,2001,22(12):56-58
88 王亚伟，李春亚.膳食纤维和低聚糖对糖尿病人血糖血脂的影响.郑州工程学院学报,2002,23(1):85-88
89王亚伟,王燕等.膳食纤维对糖尿病人血糖的影响.食品工业技,2001,22(5):25-27
90 Elias,A.N.,Grossman,M.K.,Valenta,L.J.,et al.Use of the artificial beta cell in the assessment of peripheral insulin sensitivity:effect of chromium supple-mentation in
diabetic patients. J.Gen Pharmacol.1984,15:535
91 赵晓华，李兴，宋征，杨燕等.加铬复合纤维对糖尿病大鼠治疗作用观察.营养学报,2000,4:312-316
92 姜桂荣等.富铬酵母对糖尿病人糖耐量的影响.营养学报,1988,10(3):253
93 Osamu Ezaki.The insulin-like effects of selenate in rat adipocytes.J.Biol Chem.1990,
265:1124
94 丁虹，彭仁琇，陈金和，黎七雄，汪晖，孔锐.硒、大蒜素对糖尿病小鼠降糖作用的研究.营养学报,1997,4:384-386
95 Ballard,F.J.,Tomas,F.M.,Read,L.C.,Knowles,S.E.,Owens,j.c.and Francis,
G.L.Effect of IGF-Ⅰand IGF analogs on growth during catabolic states
in rat.In Modern Concepts of Insulin-like Growth Factors.
(Edited by Spencer,E.M.)Elsevier,New York.1991,617-627
96 Read,l.c.,Lemmey,A.B.,Howarth,G.S.,Martin,A.A.,Tomas,F.M.and Ballard,
F.J.The gastrointestinal tract is one of the most responsive target
tissues forIGF-Ⅰand its potent analogs. In Modern Concepts of Insulin-like
Growth Factors.(Edited by Spencer,E.M.) Elsevier,New York.1991,225-234
97 叶红英，俞茂华，游利，颜贻谦.外源性牛初乳短链IGF-Ⅰ治疗改善糖尿病大鼠GH/ IGF-Ⅰ轴的紊乱.中国糖尿病杂志.2002,1:15-16
98 孙侃，俞茂华，史虹莉，杨秀芳，方京冲等.牛初乳短链IGF-Ⅰ对STZ糖尿病大鼠物质代谢及肾脏的影响.中华内分泌代谢杂志.1997,3:162-166
99 俞茂华,朱禧星,史虹莉,何德华等.牛初乳制剂治疗型糖尿病的临床初步观察.上海医科大学学报,1992,19(1):41-44
100 Etherton,T.D.,Wiggins,J.P.,Evock,C.M.,Chung,C.S.,Rebhun,J.F.,Walton,
P.E.& Steele,N.C. Stimulation of pig growth performance by porcine
growth hormone:Determination of the dose-response relationship.J.
Animal Science.1987,64:433-443
101 张龙翔,张庭芳,李令媛主编.生化实验方法和技术.北京:高等教育出版社,1997,136-137
102 徐叔云,卡如濂,陈修主编.药理实验方法学（第二版）.北京:人民卫生出版社,1995,1277-1278
103 H.G沃格尔[德],W.H.沃格尔[美]编著.杜冠华,李学军,张永祥等译.药理学实验指南－新药发现和药理学评价.北京:科学出版社,2001,2:700-701
104 Rao,R.K. Biologically active peptides in the gastrointestinal lumen.Life
Sci.1991,48:1685-1704