CAJ | 학술논문

本试验研究了不同浓度的β-羟丁酸( BHBA)对奶牛乳腺上皮细胞( BMEC)内乳蛋白合成相关基因表达量的影响。将传至第3代的BMEC以适宜的密度培养48 h后,随机分为6组,每组6个重复,每个重复1个培养孔。各组分别采用BHBA浓度为0(对照组)、0.58、1.16、2.32、4.64和9.28 mmol/L的培养液,置于37℃、5% CO2培养箱中继续培养48 h。收集细胞,采用SYBR Green实时荧光定量PCR法测定乳蛋白合成相关基因的表达量。结果表明：随着BHBA浓度的增加,αs1-酪蛋白( CSN1S1)基因表达量呈显著的一元二次曲线增加( R2=0.9316,P=0.018),其中以2.32~9.28 mmol/L组较高,尤以4.64 mmol/L BHBA组最高。随着BHBA浓度的增加,瘦素(leptin)基因表达量呈一定的一次线性降低趋势(R2=0.4825,P=0.126),从数值上看,以0.58 mmol/L BHBA组最高。信号转导和转录激活因子5( STAT5)和哺乳动物雷帕霉素靶点(mTOR)基因表达量与 BHBA 浓度间的回归关系不显著(P=0.459,P=0.398)。综合以上指标,BHBA的浓度为2.32~9.28 mmol/L可上调CSN1S1基因的表达,尤以BHBA浓度为4.64 mmol/L时对BMEC内乳蛋白合成的促进效果最好。
본시험연구료불동농도적β-간정산( BHBA)대내우유선상피세포( BMEC)내유단백합성상관기인표체량적영향。장전지제3대적BMEC이괄의적밀도배양48 h후,수궤분위6조,매조6개중복,매개중복1개배양공。각조분별채용BHBA농도위0(대조조)、0.58、1.16、2.32、4.64화9.28 mmol/L적배양액,치우37℃、5% CO2배양상중계속배양48 h。수집세포,채용SYBR Green실시형광정량PCR법측정유단백합성상관기인적표체량。결과표명：수착BHBA농도적증가,αs1-락단백( CSN1S1)기인표체량정현저적일원이차곡선증가( R2=0.9316,P=0.018),기중이2.32~9.28 mmol/L조교고,우이4.64 mmol/L BHBA조최고。수착BHBA농도적증가,수소(leptin)기인표체량정일정적일차선성강저추세(R2=0.4825,P=0.126),종수치상간,이0.58 mmol/L BHBA조최고。신호전도화전록격활인자5( STAT5)화포유동물뢰파매소파점(mTOR)기인표체량여 BHBA 농도간적회귀관계불현저(P=0.459,P=0.398)。종합이상지표,BHBA적농도위2.32~9.28 mmol/L가상조CSN1S1기인적표체,우이BHBA농도위4.64 mmol/L시대BMEC내유단백합성적촉진효과최호。
The purpose of this study was to examine the effects ofβ-hydroxybutyric acid ( BHBA) on expres-sions of genes involved in milk protein synthesis in bovine mammary epithelial cells ( BMECs) . The 3rd pas-sage BMECs with proper densities were cultured for 48 h, then were divided into 6 groups with 6 replicates in each group and 1 culture pore per replicate. Culture mediums containing different concentrations of BHBA [ 0 ( control group) , 0.58, 1.16, 2. 32, 4. 64 and 9. 28 mmol/L, respectively] were used in each group. Cells were continuous incubated in an incubator with 5% CO2 at 37 ℃ for 48 h, then were collected for the analysis of expressions of milk protein and the expression of related genes in real-time fluorescent quantitative PCR with SYBR Green. The results showed as follows: with the increasing of BHBA concentration, the expression of CSN1S1 gene was significantly increased quadratically ( R2=0.931 6, P=0.018) , 2.32 to 9.28 mmol/L had better effect, and 4.64 mmol/L was optimal concentration. With the increasing of BHBA concentration, leptin gene expression was linearly down-regulated at a certain extent ( R2=0.482 5, P=0.126) , and 0.58 mmol/L was optimal concentration in value. With the increasing of BHBA concentration, signal transducer and activator of transcription 5 ( STAT5) and mammalian target of rapamycin ( mTOR) gene expression were not significant-ly changed ( P=0.459, P=0.398) . Taken together, the expression of CSN1S1 gene is up-regulated when cells areincubated with BHBA at 2. 32 to 9. 28 mmol/L, and the synthesis of milk proteins in BMECs has better effect when the concentration of BHBA is 4. 64 mmol/L.