二、三維培養(yǎng)模式下添加激素對奶牛乳腺上皮細胞乳脂肪和乳蛋白合成的影響
發(fā)布時間：2018-08-23 09:30
【摘要】：本論文研究在二、三維培養(yǎng)模式下,添加不同濃度的催乳素(PRL)、胰島素(INS)、生長激素(GH)及三者不同濃度配比對奶牛乳腺上皮細胞(BMECs)乳脂肪和乳蛋白合成的影響,從乳脂肪及乳蛋白合成相關(guān)基因和轉(zhuǎn)錄調(diào)控因子基因表達水平初步探明激素調(diào)控乳脂肪和乳蛋白合成的分子機制,為調(diào)控乳成分合成提供理論依據(jù),為研究奶牛泌乳生物學機制時乳腺上皮細胞培養(yǎng)模型的選擇提供數(shù)據(jù)支撐。論文包含四個試驗,每個試驗包含兩部分研究內(nèi)容,第一部分研究在二維培養(yǎng)模式下,單獨添加PRL(0、100、300、500、1000ng/mL)、INS(0、25、100、400、800ng/mL)和 GH(0、10、30、100、300 ng/mL)以及組合添加不同配比的PRL、INS和GH[試驗Ⅰ組(0)、試驗Ⅱ組(20:1:2)、試驗Ⅲ組(7.3:1:1.8)]研究其對BMECs甘油三酯(TAG)含量和乳脂肪及乳蛋白合成相關(guān)基因表達的影響。第二部分內(nèi)容根據(jù)第一部分研究篩選出的適宜激素濃度進行單一或組合添加,同時進行BMECs的二維和三維培養(yǎng)。激素單一添加時PRL濃度為0、100和200 ng/mL,INS濃度為0、5和10 ng/mL,GH濃度為0、10和30 ng/mL。組合添加時PRL、INS和GH的總添加量相同,均為140 ng/mL,試驗Ⅰ組為對照組、試驗Ⅱ組三種激素的配比為10:1:3,試驗Ⅲ組為20:1:2,試驗Ⅳ組為2:1:1,比較不同培養(yǎng)模式下BMECs培養(yǎng)基中TAG含量以及乳脂肪和乳蛋白合成相關(guān)基因的表達差異。試驗一研究表明,二維培養(yǎng)模式下,100 ng/mL PRL顯著提高BMECs脂肪酸合成酶(FASNN)、雷帕霉素靶分子(mTOR)、核糖體p70s6激酶(S6K1)、真核翻譯啟始因子4E結(jié)合蛋白(4EBP1)、α-酪蛋白(CSN1S1)和催乳素受體(PRLR)基因的表達量。300ng/mL PRL顯著提高乙酰輔酶A羧化酶a(ACC)、二脂酰甘油酰基轉(zhuǎn)移酶2(DGAT2)、脂肪酸結(jié)合蛋白3(FABP3)、PRLR基因表達量及甘油三酯(TAG)的含量。結(jié)果提示,二維培養(yǎng)模式下100～300 ng/mL PRL促進乳脂肪和乳蛋白的合成,但是高濃度(1 000 ng/mL)的PRL對BMECs乳脂肪和乳蛋白的合成有一定抑制作用。三維培養(yǎng)模式下,培養(yǎng)基中TAG含量及BMECs乳脂肪和乳蛋白合成相關(guān)基因表達量均顯著高于二維培養(yǎng)模式。100 ng/mL組AC FASN、硬脂酰CoA 去飽和酶(SCT)、DGAT2、固醇調(diào)節(jié)元件結(jié)合蛋白1(SREBP1)、過氧化物酶體增殖物激活受體γ(PPARG)、4EBP1、β-酪蛋白(CSN2)、K-酪蛋白(CSN3)、PRLR的mRNA相對表達量及TAG合成量顯著高于對照組及200 ng/mL PRL組。200 ng/mL PRL組顯著促進絲氨酸/蘇氨酸激酶(AKT)、mTOR、CSN1S1基因表達量。結(jié)果表明,三維培養(yǎng)模式下PRL添加量為100 ng/mL時,對乳脂肪和乳蛋白的合成有較好的促進作用。試驗二研究表明,在二維培養(yǎng)模式下INS濃度為25ng/mL和100ng/mL時,BMECs TAG含量顯著高于對照組。25 ng/mL INS顯著提高ACC、SREBP1基因表達量,100 ng/mL INS顯著促進4EBP1基因表達量,顯著抑制S6K1、ACC、EASN、SCD、DGAT2、SREBP1、FABP3、STAT5、ACSN3、INSR基因表達量。結(jié)果表明,二維培養(yǎng)模式下INS添加量為0～25 ng/mL時,對乳脂肪和乳蛋白的合成有較好的促進作用,100～800 ng/mL對乳脂肪和乳蛋白的合成有抑制作用。在三維培養(yǎng)模式下,培養(yǎng)基中TAG合成量及BMECs乳脂肪和乳蛋白合成相關(guān)基因表達量均顯著高于二維培養(yǎng)模式。5 ng/mL INS顯著上調(diào)ACC、PPARG、AKT、S6K1、CSN1S1 基因表達量;10 ng/mL INS 組 FASSN、DGAT2、FABP3、4EBP1、STAT5、IANSRR基因表達量及TAG合成量顯著高于對照組及5 ng/mL組。結(jié)果表明,三維培養(yǎng)模式下INS添加量為10 ng/mL時,對乳脂肪和乳蛋白的合成有較好的促進作用。試驗三研究表明,在二維培養(yǎng)模式下,10 ng/mL GH顯著提高FASN、DGATT、SREBP1、PPARG、STAT5、CSN1S1、GHR基因表達量;30ng/mL GH顯著提高SREBP1基因表達量及TAG合成量。結(jié)果表明,二維培養(yǎng)模式下GH添加量為10～30 ng/mL時,促進BMECs乳脂肪和乳蛋白的合成,100-300 ng/mL GH對BMECs乳脂肪和乳蛋白的合成有抑制作用。在三維培養(yǎng)模式下,培養(yǎng)基中TAG合成量及BMECs乳脂肪和乳蛋白合成相關(guān)基因表達量均顯著高于二維培養(yǎng)模式。添加10 ng/mL GH顯著上調(diào)BMECs內(nèi)ACC、SCD、DGAT2、CSN2、CSN3基因表達量,顯著抑制S6K1基因表達量;30 ng/mL組BMECs內(nèi)FASN、SREBP1、PPARG、FABP3、AKT、mTOR、4EBP1、S6K1、STAT5、CSN1S1、GHR、IGFR基因表達量及TAG合成量顯著高于對照組及10ng/mL組。結(jié)果表明,在三維培養(yǎng)模式下,GH添加量為30 ng/mL時,對BMECs乳脂肪和乳蛋白的合成有較好的促進作用。試驗四研究表明,二維培養(yǎng)模式下,試驗Ⅱ組(20:1:2)DGT、FABP3、SREBP1、CSN1S1、STAT5、mTOR、AKT、4EBP1、S6K1、GHR、IGFR基因表達量及 TAG 合成量顯著高于對照組及試驗Ⅲ組(7.3:1:1.8)。結(jié)果表明,二維培養(yǎng)模式下,PRL、INS和GH組合比例為20:1:2時,對BMECs乳脂肪和乳蛋白合成有較好的促進作用。在三維培養(yǎng)模式下,培養(yǎng)基中TAG合成量及BMECs乳脂肪和乳蛋白合成相關(guān)基因表達量均顯著高于二維培養(yǎng)模式。試驗Ⅱ組(10:1:3)ACC、FASNNN、DGAT1、FABP3、CSN1S1、CSN2、CSN3和AKT基因的表達量顯著高于試驗Ⅰ組、Ⅲ組(20:1:2)和Ⅳ組(2:1:1),SCDD、PPARG、mTOR、4EBP1、S6K1和STAT5基因的表達量顯著高于試驗Ⅰ組(P0.05)。試驗Ⅱ組和Ⅲ組PRLR、INNSR、GHR和IGFR基因表達量顯著高于試驗Ⅰ組和Ⅳ組(P0.05)。試驗Ⅲ組SCD和SREBP1基因表達量顯著高于其他三組。試驗Ⅳ組4EBP1、STAT5、IGFR基因的表達量顯著高于其他三組。結(jié)果表明,三維培養(yǎng)模式下,PRL、INS和GH組合比例為10:1:3時,對BMECs乳脂肪和乳蛋白的合成有較好的促進作用。
[Abstract]:In this paper, the effects of different concentrations of prolactin (PRL), insulin (INS), growth hormone (GH) and their combinations on milk fat and milk protein synthesis in bovine mammary epithelial cells (BMECs) were studied under two-and three-dimensional culture mode. The expression levels of genes related to milk fat and milk protein synthesis and transcription regulators were preliminarily explored. To study the molecular mechanism of hormone regulating milk fat and protein synthesis and provide theoretical basis for regulating milk composition synthesis, and to provide data support for the selection of mammary gland epithelial cell culture model in studying the biological mechanism of lactation in dairy cows. Under the formula, PRL (0,100,300,500,1000ng/mL), INS (0,25,100,400,800ng/mL) and GH (0,10,30,100,300 ng/mL) were added separately and different ratios of PRL, INS and GH were added in combination [test group I (0), test group II (20:1:2), test group III (7.3:1:1.8)] to study the content of triglyceride (TAG) and the expression of genes related to milk fat and milk protein synthesis. In the second part, the optimum hormone concentration was selected according to the first part of the study. The two-dimensional and three-dimensional culture of BMECs was carried out simultaneously. The PRL concentration was 0,100 and 200 ng/mL, the INS concentration was 0,5 and 10 ng/mL, and the GH concentration was 0,10 and 30 ng/mL. The same, 140 ng / mL, test group I as the control group, test group II three hormones ratio of 10:1:3, test group III 20:1:2, test group IV 2:1:1, compare the different culture modes of BMECs medium TAG content and milk fat and milk protein synthesis related genes expression. Test 1 study, two-dimensional culture mode, 100 ng / 3 ML PRL significantly increased the expression of BMECs fatty acid synthase (FASNN), rapamycin target molecule (mTOR), ribosomal p70S6 kinase (S6K1), eukaryotic translation initiation factor 4E binding protein (4EBP1), alpha-casein (CSN1S1) and prolactin receptor (PRLR) genes. 300ng/mL PRL significantly increased the expression of ACA carboxylase (ACC), diacylglycerol transferase 2 (D-glycerol transferase 2). GAT2, FABP3, PRLR gene expression and triglyceride (TAG) content. The results suggested that 100-300 ng/mL PRL promoted the synthesis of milk fat and milk protein in two-dimensional culture mode, but high concentration (1 000 ng/mL) PRL inhibited the synthesis of milk fat and milk protein in BMECs. TAG content and BMECs milk fat and milk protein synthesis related gene expression were significantly higher than those of two-dimensional culture model. 100 ng/ml group AC FASN, stearoyl CoA desaturase (SCT), DGAT2, sterol regulatory element binding protein 1 (SREBP1), peroxisome proliferator-activated receptor gamma (PPARG), 4EBP1, beta-casein (CSN2), K-casein (CSN3), PRLR. The relative mRNA expression and TAG synthesis of PRL group were significantly higher than those of control group and 200 ng/mL PRL group. 200 ng/mL PRL group significantly promoted the expression of serine/threonine kinase (AKT), mTOR and CSN1S1 gene. The results showed that PRL supplementation of 100 ng/mL in three-dimensional culture mode could promote the synthesis of milk fat and milk protein. The results showed that the TAG content of BMECs was significantly higher than that of control group when INS concentration was 25 ng/mL and 100 ng/mL. 25 ng/mL INS significantly increased ACC, SREBP1 gene expression, 100 ng/mL INS significantly increased 4EBP1 gene expression, significantly inhibited S6K1, ACC, EASN, SCD, DGAT2, SREBP1, FABP3, STAT5, ACSN3, INSR gene expression. When the INS content was 0-25 ng/mL, it could promote the synthesis of milk fat and milk protein, and 100-800 ng/mL could inhibit the synthesis of milk fat and milk protein. 5 ng/mL INS significantly increased the expression of ACC, PPARG, AKT, S6K1, CSN1S1 genes; 10 ng/mL INS group FASSN, DGAT2, FABP3, 4EBP1, STAT5, IANSRR gene expression and TAG synthesis were significantly higher than the control group and 5 ng/mL group. Trial 3 showed that 10 ng/mL GH significantly increased the expression of FASN, DGATT, SREBP1, PPARG, STAT5, CSN1S1 and GHR genes under two-dimensional culture mode, and 30 ng/mL GH significantly increased the expression of SREBP1 gene and the synthesis of TAG. The results showed that the addition of 10-30 ng/mL GH promoted the expression of fat and milk protein in BMECs. In three-dimensional culture, TAG synthesis and expression of genes related to milk fat and milk protein synthesis in BMECs were significantly higher than those in two-dimensional culture. 10 ng/mL GH significantly increased the expression of ACC, SCD, DGAT2, CSN2 and CSN3 genes in BMECs. The expression of FASN, SREBP1, PPARG, FABP3, AKT, mTOR, 4EBP1, S6K1, STAT5, CSN1S1, GHR, IGFR and TAG synthesis in BMECs of 30 ng/mL group were significantly higher than those of control group and 10 ng/mL group. Experiment 4 showed that the expression of DGT, FABP3, SREBP1, CSN1S1, STAT5, mTOR, AKT, 4EBP1, S6K1, GHR, IGFR and TAG synthesis were significantly higher in group II (20:1:2) than in control group and group III (7.3:1:1.8). The results showed that the ratio of PRL, INS and GH was 20:1:2 for BMECs milk fat and fat. TAG synthesis and BMECs milk fat and milk protein synthesis related gene expression were significantly higher in three-dimensional culture medium than in two-dimensional culture model. ACC, FASNNN, DGAT1, FABP3, CSN1S1, CSN2, CSN 3 and AKT gene expression in test group II (10:1:3) were significantly higher than those in test group I and group III (20:1:3). The expression levels of PRLR, INNSR, GHR and IGFR genes in group II and group III were significantly higher than those in group I and group IV (P 0.05). The expression levels of SCD, PPARG, mTOR, 4EBP1, S6K 1 and STAT5 genes in group III were significantly higher than those in other three groups. The results showed that when the ratio of PRL, INS and GH was 10:1:3, the synthesis of milk fat and protein in BMECs was promoted.
【學位授予單位】：內(nèi)蒙古農(nóng)業(yè)大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：S823
本文編號：2198632
[Abstract]:In this paper, the effects of different concentrations of prolactin (PRL), insulin (INS), growth hormone (GH) and their combinations on milk fat and milk protein synthesis in bovine mammary epithelial cells (BMECs) were studied under two-and three-dimensional culture mode. The expression levels of genes related to milk fat and milk protein synthesis and transcription regulators were preliminarily explored. To study the molecular mechanism of hormone regulating milk fat and protein synthesis and provide theoretical basis for regulating milk composition synthesis, and to provide data support for the selection of mammary gland epithelial cell culture model in studying the biological mechanism of lactation in dairy cows. Under the formula, PRL (0,100,300,500,1000ng/mL), INS (0,25,100,400,800ng/mL) and GH (0,10,30,100,300 ng/mL) were added separately and different ratios of PRL, INS and GH were added in combination [test group I (0), test group II (20:1:2), test group III (7.3:1:1.8)] to study the content of triglyceride (TAG) and the expression of genes related to milk fat and milk protein synthesis. In the second part, the optimum hormone concentration was selected according to the first part of the study. The two-dimensional and three-dimensional culture of BMECs was carried out simultaneously. The PRL concentration was 0,100 and 200 ng/mL, the INS concentration was 0,5 and 10 ng/mL, and the GH concentration was 0,10 and 30 ng/mL. The same, 140 ng / mL, test group I as the control group, test group II three hormones ratio of 10:1:3, test group III 20:1:2, test group IV 2:1:1, compare the different culture modes of BMECs medium TAG content and milk fat and milk protein synthesis related genes expression. Test 1 study, two-dimensional culture mode, 100 ng / 3 ML PRL significantly increased the expression of BMECs fatty acid synthase (FASNN), rapamycin target molecule (mTOR), ribosomal p70S6 kinase (S6K1), eukaryotic translation initiation factor 4E binding protein (4EBP1), alpha-casein (CSN1S1) and prolactin receptor (PRLR) genes. 300ng/mL PRL significantly increased the expression of ACA carboxylase (ACC), diacylglycerol transferase 2 (D-glycerol transferase 2). GAT2, FABP3, PRLR gene expression and triglyceride (TAG) content. The results suggested that 100-300 ng/mL PRL promoted the synthesis of milk fat and milk protein in two-dimensional culture mode, but high concentration (1 000 ng/mL) PRL inhibited the synthesis of milk fat and milk protein in BMECs. TAG content and BMECs milk fat and milk protein synthesis related gene expression were significantly higher than those of two-dimensional culture model. 100 ng/ml group AC FASN, stearoyl CoA desaturase (SCT), DGAT2, sterol regulatory element binding protein 1 (SREBP1), peroxisome proliferator-activated receptor gamma (PPARG), 4EBP1, beta-casein (CSN2), K-casein (CSN3), PRLR. The relative mRNA expression and TAG synthesis of PRL group were significantly higher than those of control group and 200 ng/mL PRL group. 200 ng/mL PRL group significantly promoted the expression of serine/threonine kinase (AKT), mTOR and CSN1S1 gene. The results showed that PRL supplementation of 100 ng/mL in three-dimensional culture mode could promote the synthesis of milk fat and milk protein. The results showed that the TAG content of BMECs was significantly higher than that of control group when INS concentration was 25 ng/mL and 100 ng/mL. 25 ng/mL INS significantly increased ACC, SREBP1 gene expression, 100 ng/mL INS significantly increased 4EBP1 gene expression, significantly inhibited S6K1, ACC, EASN, SCD, DGAT2, SREBP1, FABP3, STAT5, ACSN3, INSR gene expression. When the INS content was 0-25 ng/mL, it could promote the synthesis of milk fat and milk protein, and 100-800 ng/mL could inhibit the synthesis of milk fat and milk protein. 5 ng/mL INS significantly increased the expression of ACC, PPARG, AKT, S6K1, CSN1S1 genes; 10 ng/mL INS group FASSN, DGAT2, FABP3, 4EBP1, STAT5, IANSRR gene expression and TAG synthesis were significantly higher than the control group and 5 ng/mL group. Trial 3 showed that 10 ng/mL GH significantly increased the expression of FASN, DGATT, SREBP1, PPARG, STAT5, CSN1S1 and GHR genes under two-dimensional culture mode, and 30 ng/mL GH significantly increased the expression of SREBP1 gene and the synthesis of TAG. The results showed that the addition of 10-30 ng/mL GH promoted the expression of fat and milk protein in BMECs. In three-dimensional culture, TAG synthesis and expression of genes related to milk fat and milk protein synthesis in BMECs were significantly higher than those in two-dimensional culture. 10 ng/mL GH significantly increased the expression of ACC, SCD, DGAT2, CSN2 and CSN3 genes in BMECs. The expression of FASN, SREBP1, PPARG, FABP3, AKT, mTOR, 4EBP1, S6K1, STAT5, CSN1S1, GHR, IGFR and TAG synthesis in BMECs of 30 ng/mL group were significantly higher than those of control group and 10 ng/mL group. Experiment 4 showed that the expression of DGT, FABP3, SREBP1, CSN1S1, STAT5, mTOR, AKT, 4EBP1, S6K1, GHR, IGFR and TAG synthesis were significantly higher in group II (20:1:2) than in control group and group III (7.3:1:1.8). The results showed that the ratio of PRL, INS and GH was 20:1:2 for BMECs milk fat and fat. TAG synthesis and BMECs milk fat and milk protein synthesis related gene expression were significantly higher in three-dimensional culture medium than in two-dimensional culture model. ACC, FASNNN, DGAT1, FABP3, CSN1S1, CSN2, CSN 3 and AKT gene expression in test group II (10:1:3) were significantly higher than those in test group I and group III (20:1:3). The expression levels of PRLR, INNSR, GHR and IGFR genes in group II and group III were significantly higher than those in group I and group IV (P 0.05). The expression levels of SCD, PPARG, mTOR, 4EBP1, S6K 1 and STAT5 genes in group III were significantly higher than those in other three groups. The results showed that when the ratio of PRL, INS and GH was 10:1:3, the synthesis of milk fat and protein in BMECs was promoted.
【學位授予單位】：內(nèi)蒙古農(nóng)業(yè)大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：S823
【參考文獻】
相關(guān)期刊論文 前10條
1 李勝利;姚琨;曹志軍;劉長全;張勝利;劉建新;李建喜;王加啟;張和平;;2016年奶牛產(chǎn)業(yè)技術(shù)發(fā)展報告[J];中國畜牧雜志;2017年01期
2 王皓宇;秦彤;郝海生;杜衛(wèi)華;趙學明;朱化彬;;胰島素對體外培養(yǎng)奶牛乳腺上皮細胞乳蛋白、乳脂肪合成相關(guān)基因mRNA表達的影響[J];畜牧獸醫(yī)學報;2013年05期
3 田青;季昀;龐學燕;王洪榮;;胰島素對奶牛乳腺上皮細胞酪蛋白合成調(diào)節(jié)機理的研究[J];動物營養(yǎng)學報;2013年03期
4 陳靜;王根林;李蓮;于靜;趙士杰;蔣小強;王斐;;雌激素對奶牛乳腺上皮細胞增殖及抗氧化性的影響[J];農(nóng)業(yè)生物技術(shù)學報;2013年02期
5 田青;王洪榮;;胰島素、催乳素和氫化可的松對奶牛乳腺上皮細胞增殖和凋亡的影響[J];中國飼料;2013年02期
6 田青;季昀;龐學燕;王洪榮;;胰島素對奶牛乳腺上皮細胞生長及κ-酪蛋白和胰島素受體基因表達的影響[J];動物營養(yǎng)學報;2013年01期
7 李冬輝;林葉;高學軍;王春梅;張莉;李慶章;;催乳素、雌激素和胰島素對奶牛乳腺上皮細胞中瘦素及其受體的影響[J];畜牧與獸醫(yī);2012年S1期
8 季昀;龐學燕;田青;王夢芝;王洪榮;敖長金;;生長激素和胰島素樣生長因子Ⅰ對奶牛乳蛋白合成關(guān)鍵激酶及調(diào)節(jié)因子mRNA表達量的影響[J];動物營養(yǎng)學報;2013年01期
9 林先滋;羅軍;張犁蘋;朱江江;石恒波;茍德明;;miR-200a對奶山羊乳腺上皮細胞乳脂合成相關(guān)基因mRNA表達的影響[J];畜牧獸醫(yī)學報;2012年07期
10 劉暢;趙鋒;李慶章;;不同泌乳相關(guān)激素和生長因子對奶牛乳腺上皮細胞增殖的影響[J];中國畜牧獸醫(yī);2012年05期
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