本文選題：乳酸發(fā)酵 + 乳酸鈣��； 參考：《內(nèi)蒙古大學(xué)》2017年碩士論文

【摘要】：響應(yīng)自治區(qū)提出的綠色農(nóng)畜產(chǎn)品輸出基地建設(shè)與轉(zhuǎn)型升級(jí)的需要,開發(fā)綠色無污染的高科技高端產(chǎn)品、增加畜產(chǎn)品附加值已成為當(dāng)務(wù)之急。以牛羊鮮骨為原料通過發(fā)酵研發(fā)補(bǔ)鈣又促成骨的純天然高端產(chǎn)品,提高畜牧資源的附加值,為民生健康做出貢獻(xiàn)。選用三種乳酸菌發(fā)酵牛羊鮮骨粉,使羥磷灰石的鈣以乳酸鈣形式釋放。經(jīng)牛羊鮮骨—粉碎—滅菌—發(fā)酵—總混—凍干—干法制粒—壓片制成生物活性制劑(產(chǎn)品),并在細(xì)胞,組織和動(dòng)物水平上驗(yàn)證了產(chǎn)品的功效。用濕灰法消化骨粉用EDTA滴定方法測(cè)定總鈣含量,用EDTA滴定方法測(cè)定游離鈣,以平板傾注法測(cè)定活菌數(shù)。培養(yǎng)人成骨細(xì)胞hFOB并用產(chǎn)品刺激12 h后提取RNA在以熒光定量PCR檢測(cè)COLLAGEN,RANKL,OPG三種基因的表達(dá)。以切除卵巢的方法制作了骨質(zhì)疏松模型動(dòng)物,并用全自動(dòng)生化儀測(cè)定血清指標(biāo),取股骨制作病理切片經(jīng)HE染色觀察大鼠骨質(zhì)疏松程度。灌胃給藥法研究個(gè)體水平上的功效,灌胃兩個(gè)月后動(dòng)脈取血測(cè)定血清生化指標(biāo),再處死大鼠后取股骨制作病理切片HE染色觀察大鼠攝取不同鈣產(chǎn)品后骨質(zhì)疏松程度。目前我們的產(chǎn)品已將不能吸收的羥磷灰石的37.61%轉(zhuǎn)化為可吸收的游離鈣(乳酸鈣)形式,即,游離鈣的含量增加了近20倍。并在骨質(zhì)疏松動(dòng)物模型動(dòng)物水平上證明了補(bǔ)鈣又能促進(jìn)骨形成。還在細(xì)胞水平也初步證明該產(chǎn)品具有增加骨保護(hù)素表達(dá)和成骨作用。因此,本研究對(duì)于鈣補(bǔ)充、促進(jìn)骨形成及預(yù)防骨質(zhì)疏松癥都具有重要意義。結(jié)論1、乳酸菌發(fā)酵有效游離出骨粉中以羥磷灰石形式存在的結(jié)合鈣為乳酸鈣,乳酸鈣含量增加了近20倍;2、增加成骨細(xì)胞外鈣離子濃度使OPG表達(dá)量增加,PANKL表達(dá)量增加,對(duì)GOLLLAGEN的表達(dá)沒有影響,說明產(chǎn)品具有成骨作用;3、通過在細(xì)胞,組織和動(dòng)物水平上實(shí)驗(yàn)證明產(chǎn)品對(duì)骨質(zhì)疏松癥的修復(fù)有顯著性的效果;我們研發(fā)產(chǎn)品的性能測(cè)定結(jié)果顯示,該產(chǎn)品具有產(chǎn)業(yè)化開發(fā)的前景。
[Abstract]:In response to the needs of the construction and transformation of the export base of green agricultural and livestock products proposed by the autonomous region, it is urgent to develop the green, non-polluting, high-tech and high-end products and to increase the added value of livestock products. Using fresh bone of cattle, sheep and sheep as raw material, calcium supplement and pure high-end product of bone can be developed through fermentation, which can improve the added value of animal husbandry resources and contribute to people's livelihood and health. The calcium of hydroxyapatite was released in the form of calcium lactate by using three kinds of lactic acid bacteria to ferment fresh bone powder of cattle and sheep. The bioactive preparation (product) was prepared by bovine and sheep fresh bone, crushing, sterilizing, fermenting, total mixing, freeze-drying, dry granulation and pressing tablets, and the efficacy of the product was verified at cell, tissue and animal levels. Total calcium content was determined by EDTA titration method, free calcium content was determined by EDTA titration method, and live bacteria number was determined by plate pour method. Human osteoblast hFOB was cultured and stimulated for 12 h, then RNA was extracted. The expression of three genes of COLLAGEN RANKL OPG was detected by fluorescence quantitative PCR. Osteoporosis model animal was made by ovariectomized method. Serum indexes were measured by automatic biochemical instrument. Pathological sections of femur were made by HE staining to observe the degree of osteoporosis in rats. The efficacy of intragastric administration was studied at individual level. The serum biochemical indexes were measured by arterial blood collection two months after gastric perfusion and then the femur was taken to make pathological sections by HE staining to observe the degree of osteoporosis after different calcium products were ingested in rats. At present, our products have converted 37.61% of unabsorbable hydroxyapatite into absorbable free calcium (calcium lactate), that is, the content of free calcium has increased nearly 20 times. At the animal level of osteoporosis animal model, calcium supplementation can promote bone formation. It was also proved that the product could increase osteoprotegerin expression and osteogenesis. Therefore, this study is important for calcium supplementation, bone formation and osteoporosis prevention. Conclusion 1. The calcium bound in hydroxyapatite was the calcium lactate in lactic acid bacteria. The content of calcium lactate increased by 20 times, and the concentration of extracellular calcium increased the expression of OPG and PANKL. There is no effect on the expression of GOLLLAGEN, which indicates that the product has osteogenic effect. It is proved by experiments on cell, tissue and animal level that the product has remarkable effect on the repair of osteoporosis. The product has the prospect of industrialization development.
【學(xué)位授予單位】：內(nèi)蒙古大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：TS251.94;R580

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