绿原酸影响酮病奶牛乳腺细胞的CPT1A和SOCE的表达  

Chlorogenic acid increases CPT1Aand SOCE in mammary cells of cows in ketosis

作　　者：王晶晶 李巳晨 李雨坤 吴狄 李铭 温佳楠 王爽[2] 徐闯 杨威[2] 张冰冰[1] WANG Jingjing;LI Sichen;LI Yukun;WU Di;LI Ming;WEN Jianan;WANG Shuang;XU Chuang;YANG Wei;ZHANG Bingbing(College of Life Science and Technology,Heilongjiang Bayi Agricultural University,Daqing,Heilongjiang163319,China;College of Animal Science and Veterinary Medicine,Heilongjiang Bayi Agricultural University,Daqing,Heilongjiang163319,China;College of Veterinary Medicine,China Agricultural University,Beijing100191,China)

机构地区：[1]黑龙江八一农垦大学生命科学技术学院,黑龙江大庆163319 [2]黑龙江八一农垦大学动物科技学院,黑龙江大庆163319 [3]中国农业大学动物医学院,北京100191

出　　处：《中国兽医学报》2024年第4期808-814,共7页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(32172926,U20A2062)。

摘　　要：运用荧光定量PCR和Western blot方法检测健康、酮病奶牛乳腺组织中肉碱棕榈酰转移酶1A(carnitine palmitoyltransferase 1a,CPT1A)及储存操作Ca2+通道(store-operated Ca2+entry,SOCE)通路相关分子Orai1、Orai3、STIM1和STIM2的表达水平;在体外乳腺细胞培养过程中,细胞处理分为对照组、BHBA组、CGA组、CGA+BHBA组。使用20mg/L绿原酸(chlorogenic acid,CGA)刺激细胞12h后用1.2mmol/L BHBA刺激细胞24h,运用荧光定量PCR和Western blot方法检测不同处理细胞中CPT1A及SOCE通路相关分子Orai1、Orai3、STIM1和STIM2的表达水平。结果显示,与健康奶牛相比,酮病奶牛乳腺组织中CPT1A的转录水平和表达水平均极显著降低(P<0.01),而Orai1、Orai3、STIM1和STIM2的转录水平和表达水平均显著升高(P<0.05)。与对照组相比,BHBA刺激降低了CPT1A的转录水平和表达水平(P<0.01),升高了Orai1、Orai3、STIM1和STIM2的转录水平和表达水平(P<0.05);与对照组相比,CGA组CPT1A、Orai1、Orai3、STIM1和STIM2的转录水平和表达水平无明显变化(P>0.05);与BHBA组相比,使用CGA和BHBA共同刺激使CPT1A的转录水平和表达水平升高(P<0.05),Orai1、Orai3、STIM1和STIM2的转录水平和表达水平降低P<0.05)。结果表明,绿原酸增加酮病奶牛乳腺细胞的CPT1A的表达并降低SOCE的表达。This experiment is to explore whether chlorogenic acid(CGA)affects the oxidation and SOCE expression of mammary cells in ketosis dairy cows.First,the expression levels of CPT1A and SOCE pathway-related molecules Orai1,Orai3,STIM1and STIM2in mammary tissue of healthy and ketosis dairy cows were detected by fluorescent quantitative PCR and western blot.The four groups used for experiment were the control group,BHBA group,CGA group,and CGA+BHBA group.The cells were stimulated with 20mg/L CGA for 12hand then stimulated with 1.2 mmol/L BHBA for 24h.The expression of the molecules Orai1,Orai3,STIM1and STIM2linked to the CPT1Aand SOCE pathways in various treated cells was assessed using fluorescent quantitative PCR and western blot.Compared with healthy cows,the transcript level and expression level of CPT1Ain the mammary gland tissue of ketosis cows were significantly decreased(P<0.01),while the transcript levels and expression levels of Orai1,Orai3,STIM1 and STIM2 were significantly increased(P<0.05).CPT1Atranscription and expression levels were reduced by BHBA stimulation in comparison to the control group(P<0.01),and increased Orai1,Orai3,STIM1and Transcript level and expression level of STIM2(P<0.05).The transcript levels and expression levels of CPT1A,Orai1,Orai3,STIM1and STIM2in the CGA group did not differ significantly from those in the control group(P>0.05).The expression level increased(P<0.05),and the transcript level and expression level of Orai1,Orai3,STIM1and STIM2decreased(P<0.05).The results showed that chlorogenic acid could increase the expression of CPT1Aand decrease the expression of SOCE in mammary cells of cows in ketosis.

关 键 词：SOCE 绿原酸 氧化 酮病 奶牛 

分 类 号：S852.3[农业科学—基础兽医学] S856[农业科学—兽医学]