通草提取物对奶牛乳腺上皮细胞乳糖合成及相关基因表达的影响

为探讨通草对奶牛乳腺上皮细胞乳糖合成及相关基因表达的影响,采用不同剂量通草提取物处理奶牛乳腺上皮细胞,利用四甲基偶氮唑盐(MTT)法检测乳腺上皮细胞增殖能力,乳糖/半乳糖检测试剂盒(Lactose/D-Galactose (Rapid) Assay Kit)检测乳腺上皮细胞培养液中乳糖含量,实时荧光定量PCR技术检测乳腺上皮细胞乳糖合成相关基因葡萄糖转运蛋白1(GLUT1)、葡萄糖转运蛋白4(GLUT4)、葡萄糖转运蛋白8(GLUT8)、葡萄糖转运蛋白12(GLUT12)、己糖激酶Ⅰ(HKⅠ)、己糖激酶Ⅱ(HKⅡ)、β-1,4-半乳糖基转移酶-1(β-4GALT1)和α-乳清白蛋白(α-LA)基因mRNA表达水平。结果表明,200、400、600μg(生药)·mL-1通草提取物提高奶牛乳腺上皮细胞增殖能力(P<0.05),促进奶牛乳腺上皮细胞合成分泌乳糖(P<0.05),并显著上调细胞GLUT1、GLUT8、HKⅡ、β-4GALT1及α-LA mRNA表达水平(P<0.05),但对GLUT4、GLUT12和HKⅠmRNA表达水平无显著影响(P>0.05)。研究表明,适当浓度通草提取物可提高奶牛乳腺上皮细胞增殖能力,并通过上调乳腺上皮细胞GLUT1、GLUT8、HKⅡ、β-4GALT1和α-LA的mRNA表达,促进乳腺上皮细胞合成乳糖。

Effect of Tetrapanax papyriferus on lactose synthesis and expression of genes related to lactose synthesis in bovine mammary epithelial cells

To investigate the effect of Tetrapanax papyriferus on lactose synthesis and the expression of genes related to lactose synthesis in bovine mammary epithelial cells. In this experiment,the bovine mammary epithelial cells were treated with different doses of Tetrapanax papyriferus extracts. The proliferation abilities of bovine mammary epithelial cells were detected by MTT method.Lactose/D-galactose(Rapid) Assay Kit was used to determine the contents of lactose in cell culture medium of mammary epithelial cells. The mRNA expression levels of genes related to lactose synthesis, such as GLUT1, GLUT4, GLUT8, GLUT12, HKⅠ, HKⅡ, β-4 GALT1, and α-LA in mammary epithelial cells were detected by q RT-PCR. The results displayed that 200, 400 and 600 μg(Crude drug) · mL-1 Tetrapanax papyriferus extracts significantly increased the proliferation ability of bovine mammary epithelial cells(P<0.05). 200, 400 and 600 μg(Crude drug) · mL-1 Tetrapanax papyriferus extracts obviously promoted the synthesis and secretion of lactose in bovine mammary epithelial cells(P<0.05), and substantially upregulated the mRNA expression levels of GLUT1, GLUT8, HK Ⅱ, β-4 GALT1, and α-LA in bovine mammary epithelial cells(P<0.05). But the mRNA expression levels of GLUT4, GLUT12 and HKⅠ were not significantly effected by Tetrapanax papyriferus extracts(P>0.05).This study showed that Tetrapanax papyriferus extract of appropriate concentration could increase the proliferation ability of bovine mammary epithelial cells and it could promote lactose synthesis through upregulating the mRNA expression of GLUT1, GLUT8, HKⅡ, β-4 GALT1, and α-LA in bovine mammary epithelial cells.