摘要
【目的】探究苦参碱对体外培养的奶牛乳腺上皮细胞(BMECs)增殖、凋亡及抗氧化能力的影响。【方法】利用含0(A组),25(B组),50(C组),75(D组)和100μg/mL(E组)苦参碱的培养基培养奶牛乳腺上皮细胞。通过四甲基偶氮唑盐(MTT)法检测BMECs活性,采用流式细胞仪(AnnexinV/PI双染法)检测苦参碱对BMECs凋亡的影响,并检测苦参碱对BMECs抗氧化酶活性及丙二醛(MDA)含量的影响,采用real-time PCR对BMECs中Caspase-3、p53、STAT1和SOCS3基因的相对表达量进行检测。【结果】用药5d时,低质量浓度(25和50μg/mL)苦参碱对BMECs增殖具有促进作用,高质量浓度(75和100μg/mL)苦参碱对细胞增殖具有抑制作用;B^E组BMECs的凋亡率均极显著高于A组(P<0.01);B^E组BMECs培养上清液中NO和乳酸脱氢酶(LDH)水平明显高于A组。B^E组BMECs的过氧化氢酶(CAT)活性均比A组高,其中C组极显著高于A组(P<0.01);B^E组的谷胱甘肽过氧化物酶(GSH-Px)活性均极显著高于A组(P<0.01),E组的超氧化物歧化酶(SOD)水平极显著高于A组(P<0.01),各组MDA含量无显著性差异。与A组相比,苦参碱上调了B^E组BMECs中Caspase-3、p53、STAT1和SOCS3基因的相对表达量。【结论】低质量浓度苦参碱能够促进BMECs增殖,高质量浓度苦参碱则会抑制BMECs增殖;不同质量浓度苦参碱均可提高BMECs的抗氧化能力,其中50μg/mL苦参碱提高BMECs抗氧化能力的效果最明显。
【Objective】This experiment was conducted to study the effect of matrine on proliferation,apoptosis and antioxidant ability of bovine mammary epithelial cells cultured in vitro.【Method】Groups A,B,C,D and E were set up and treated with matrine at concentrations of 0,25,50,75 and 100μg/mL,respectively.The activity of BMECs was detected by MTT assay.The effect of matrine on apoptosis of BMECs was detected by flow cytometry(Annexin V/PI double staining).The effect on antioxidase activityand malondialdehyde(MDA)content were also detected.The relative expressions of Caspase-3,p53,STAT1 and SOCS3 in BMECs were detected by real-time PCR.【Result】Low concentrations of matrine(25 and 50μg/mL)promoted cell proliferation after 5 days,while high concentrations(75 and 100μg/mL)inhibited cell proliferation.The apoptosis rates of groups B-E were significantly higher than that of group A(P〈0.01).The levels of NO and LDH of groups B-E were significantly higher than those of group A(P〈0.01).The CAT activities of groups B-E were higher than that of group A and there was significant difference between group C and group A(P〈0.01).GSH-Px activities of groups B-E were significantly higher than that of group A(P〈0.01).The SOD level of group E was significantly higher than that of group A(P〈0.01).There was no significant difference in MDA content.Compared with group A,matrine increased the relative expressions of Caspase-3,p53,STAT1,and SOCS3 in experimental groups.【Conclusion】Low matrine concentrations promoted the proliferation of bovine mammary epithelial cells,while high concentrations inhibited cell proliferation and increased the antioxidant abilities of BMECs.Group C with marine concentration of 50μg/mL was the most effective.
作者
卢金霞
何芳
冯峰
管翠萍
刘军红
周学章
LU Jinxia;HE Fang;FENG Feng;GUAN Cuiping;LIU Junhong;ZHOU Xuezhang(Key Lab of Ministry of Education for Protection and Utilization of Special Biological Resources in Western China Ningxia University,Yinchuan,Ningxia 750021,China;College of Veterinary Medicine,Northwest A& F University,Yangling,Shaanxi 712100,China)
出处
《西北农林科技大学学报(自然科学版)》
CSCD
北大核心
2018年第7期1-6,14,共7页
Journal of Northwest A&F University(Natural Science Edition)
基金
国家自然科学基金项目(31460676)