SIRT3对棕榈酸诱导的奶牛乳腺上皮细胞凋亡的影响
Effects of SIRT3 on palmitic acid-induced apoptosis in bovine mammary epithelial cells
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摘要: 为了研究腺病毒(Ad)介导的沉默信息调控因子2样蛋白3(sirtuin 3,SIRT3)基因过表达对棕榈酸(palmitic acid, PA)诱导的奶牛乳腺上皮细胞(bovine mammary epithelial cells, BMEC)凋亡的影响,试验将复苏的BMEC分为空白对照组(NC组)、PA组、Ad-SIRT3+PA组和Ad-Green Fluorescent Proteins(GFP)+PA组,进行相应处理后运用流式细胞术检测细胞凋亡水平,并采用Western-blot检测细胞凋亡抑制因子B淋巴细胞瘤-2(B-cell lymphoma-2,Bcl-2)蛋白、Bcl-2关联X蛋白(Bcl-2-associated X protein, Bax)和SIRT3蛋白表达水平。结果表明:与NC组相比,PA处理极显著提高了BMEC的凋亡率(P<0.01),且可极显著增加促凋亡蛋白Bax表达水平(P<0.01)、极显著降低抑制凋亡蛋白Bcl-2和SIRT3表达水平(P<0.01);与Ad-GFP+PA组相比,Ad-SIRT3+PA组凋亡率显著下降(P<0.05),Bax蛋白表达水平极显著下降(P<0.01),而Bcl-2蛋白表达水平极显著升高(P<0.01)。说明SIRT3能够缓解PA诱导的BMEC凋亡,具有缓解围生期奶牛血液高浓度非酯化脂肪酸(nonesterified fatty acid, NEFA)对乳腺脂毒性的潜质。Abstract: In order to study the effect of adenovirus(AD)-mediated silencing information regulator sirtuin 3(SIRT3) gene overexpression on palmitic acid(PA)-induced apoptosis of bovine mammary epithelial cells(BMEC), the resuscitated BMEC were divided into control group(NC group), PA group, Ad-SIRT3+PA group and Ad-green Fluorescent protein(GFP)+PA group. After corresponding treatment, cell apoptosis levels were detected by flow cytometry, and the expression levels of anti-apoptotic protein B-cell lymphoma-2(Bcl-2), pro-apoptotic protein Bcl-2-associated X protein(Bax) and SIRT3 were measured by the Western-blot assay. The results showed that compared with NC group, PA treatment significantly increased the apoptosis rate of BMEC(P<0.01) and the expression level of Bax(P<0.01), and significantly decreased the expression level of Bcl-2 and SIRT3(P<0.01). Compared with Ad-GFP+PA group, the apoptosis rate of Ad-SIRT3+PA group was significantly decreased(P<0.05), the expression level of Bax protein was extremely significantly decreased(P<0.01), while the expression level of Bcl-2 protein was extremely significantly increased(P<0.01). It indicated that SIRT3 could alleviate the apoptosis of BMEC induced by PA, and had the potential to mitigate perinatal cow blood nonesterified fatty acid(NEFA) toxicity to mammary lipid.