大豆蛋白之胰蛋白酶水解物及其純化胜肽對卵巢切除誘導骨質疏鬆症小鼠之改善作用

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Please use this identifier to cite or link to this item: http://140.128.103.80:8080/handle/310901/31648

Title:	大豆蛋白之胰蛋白酶水解物及其純化胜肽對卵巢切除誘導骨質疏鬆症小鼠之改善作用
Other Titles:	Ameliorating effect of trypsin-soy protein hydrolysate and its isolated peptide on ovariectomized-induced osteoporosis mice
Authors:	張文彥 CHANG,WEN-YEN
Contributors:	江文德;蔣慎思 CHIANG,WEN-DEE;CHIANG,SHEN-SHIH 食品科學系
Keywords:	胜肽;大豆蛋白;骨質疏鬆;鈣 peptide;soy protein;osteoporosis;calcium
Date:	2019
Issue Date:	2019-12-16T04:00:11Z (UTC)
Abstract:	本研究室先前研究成果指出以Trypsin水解分離大豆蛋白，所得到水解物(Isolated soy protein hydrolysate; IH)具有促鈣吸收之作用，從其分離出胜肽序列為NLQGENEEEDSGAIVTVK (NL)，故本研究將探討NL體內促鈣吸收的功能；首先探討口服鈣對於控制組小鼠(CON)及卵巢切除誘導骨質疏鬆(OVX)小鼠骨質影響，結果顯示OVX小鼠骨小樑骨體積比 (Bone volume / Tissue volume；BV/TV) 及骨小樑數目(Trabecular number；Tb.N) 顯著低於健康小鼠，而骨小樑分離度(Trabecular separation；Tb.Sp )顯著上升(p < 0.05)，在口服碳酸鈣8週後BV/TV、Tb.N及Tb.Sp並無顯著改善效果，健康小鼠骨小樑以桿狀為主，因此骨小樑模式因子(Trabecular bone pattern factor；TB.Pf)及結構模型指數(Structure model index；SMI)結果中CON組數值顯著高於其他組別。其次為添加不同劑量水解物IH及NL對OVX小鼠骨質影響，探討其對於骨質疏鬆症改善作用。將小鼠隨機分成10組，每組8隻包含卵巢切除處理組(OVX)、OVX加鈣組(OVX+ CaCO3；OC)、未水解ISP 200 mg/kg加鈣組(ISP+ CaCO3；ISPC)、低劑量100 mg/kg IH加鈣組(LIH+ CaCO3；LIHC)、中劑量200 mg/kg IH加鈣組(MIH+ CaCO3；MIHC)、高劑量400 mg/kg IH加鈣組(HIH+ CaCO3；HIHC)、低劑量50 mg/kg NL加鈣組(LNL+ CaCO3；LNLC)、中劑量100 mg/kg NL加鈣組(MNL+ CaCO3；MNLC)及高劑量200 mg/kg NL加鈣組(HNL+ CaCO3；HNLC)及對照組酪蛋白磷酸胜肽200 mg/kg加鈣組 (CPP+ CaCO3；CPPC)。結果表示餵食不同劑量IH及NL後，LNLC、MNLC、HNLC可顯著提升BV/TV，MNLC對於增加骨小樑數目有最佳效果，LNLC、MNLC及低中高劑量IHC均可有效降低骨小樑之間距離(p < 0.05)，其結果與正控制組CPPC相比無顯著差異，表示樣品與CPPC之間具有相同效果，其中又以MNLC具有最佳改善骨質疏鬆之效果。綜合上述結果，單純口服碳酸鈣對於骨質疏鬆症小鼠骨質並無顯著改善效果，表示碳酸鈣對骨質改善效果有限，在餵食不同劑量大豆蛋白水解物及其區分物NL合成胜肽後，可以促進小鼠體內鈣之吸收，有效增加骨小樑數目及縮短骨小樑之間距離，大豆蛋白水解物具有改善因更年期而引起骨質疏鬆症的保健食品開發潛力。 In previous studies from our lab it was demonstrated that hydrolysis of isolated soy protein (ISP) produced ISP hydrolysate (IH) with calcium-absorption stimulating (CAS) activity. The CAS peptide was identified as NLQGENEEEDS-GAIVTVK (NL). Therefore, in this study ameliorating effect of NL on osteoporosis was investigated in ovariectomized (OVX) mice. The first part was to compare oral calcium for healthy mice (CON) and OVX mice on trabecular parameter. The results showed that trabecular bone volume (Bone volume/Tissue volume; BV/TV) and trabecular number (Trabecular number; Tb.N) of OVX mice were significantly lower than those of CON group, whereas trabecular separation (Trabecular separation; Tb.Sp ) significantly increased than CON gorup (p < 0.05). BV/TV, Tb.N and Tb.Sp did not significantly improve after 8 weeks of oral calcium carbonate. The bone structure of CON group was mainly rod-shaped. Trabecular bone pattern factor (TB.Pf) and structure model index (SMI) values of CON group were significantly higher than those of other groups. The second part was to explore the effects of different doses of hydrolysate IH and NL on trabecular parameter of OVX mice. The mice were randomly divided into 10 groups, including ovariectomy group (OVX), OVX with calcium group (OVX+ CaCO3; OC), unhydrolyzed ISP 200 mg/kg with calcium group (ISP+ CaCO3; ISPC), low dose 100 mg/kg IH with calcium group (LIH+ CaCO3; LIHC), medium dose 200 mg/kg IH with calcium group (MIH+ CaCO3; MIHC), high dose 400 mg/kg IH with calcium group (HIH+CaCO3; HIHC), low dose 50 mg/kg NL with calcium group (LNL+ CaCO3; LNLC), medium dose 100 mg/kg NL with calcium group (MNL+ CaCO3; MNLC) and high dose 200 mg/kg NL with calcium group ( HNL+ CaCO3; HNLC) and the casein phosphopeptide peptide 200 mg/kg with calcium group (CPP+ CaCO3; CPPC). The results indicated that BV/TV was significantly improved after oral LNLC, MNLC and HNLC. The distance between trabecular bone was significantly reduced after oral any dosage of IHC, LNLC and MNLC, respectively, which compare to positive control CPPC. Among them, MNLC showed the highest ameliorating effect on osteoporosis. Based on the above results, only oral calcium carbonate had no significant improving effect on bone quality of osteoporosis mice, indicating that calcium carbonate had limited effect on bone quality. However, oral different dosage of IH and NL combined with calcium could promote absorption of calcium, effectively increase Tb.N and decrease Tb.Sp. Soy protein hydrolysate showed the potential to apply in the development of health food for osteoporosis after menopause.
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