摘要
为了探明绿原酸(CGA)在围产期奶牛肝细胞脂沉积的治疗作用,本试验提出CGA抑制内质网应激(ERS)缓解高游离脂肪酸(FAs)引起犊牛肝细胞脂沉积的科学假说。体外分离培养原代犊牛肝细胞添加刺激并分组,添加FAs刺激的细胞分为4组,分别为CTRL组、FAs组、CGA组、CGA+FAs组。添加毒胡萝卜素(thapsigargin,TG)刺激的细胞也分为4组,分别为CTRL组、TG组、CGA组、CGA+TG组。通过CGA对TG诱导ERS的缓解作用可以验证其影响犊牛肝细胞脂沉积的途径。样品分组后拟运用分子生物学和细胞生物学等技术,检测各组ERS指标(ATF6、PERK、IREI、CHOP、GRP78)、脂合成指标(FAS、SREBP、ACC1)和脂滴形态分布。结果表明,添加FAs与TG刺激后,犊牛肝细胞脂滴分布呈上升趋势,ERS指标(ATF6、PERK、IREI、CHOP、GRP78)、脂合成指标(FAS、SREBP、ACC1)的蛋白表达量升高,而添加CGA后,两种刺激组的脂滴分布、ERS指标(ATF6、PERK、IREI、CHOP、GRP78)、脂合成指标(FAS、SREBP、ACC1)的蛋白水平较刺激组均有缓解。可以看出CGA可以缓解由FAs引起的脂沉积,并且CGA对TG诱导的ERS起到缓解作用,因此CGA抑制ERS减缓高FAs引起的犊牛肝细胞脂沉积。
To explore the therapeutic effect of chlorogenic acid(CGA)on hepatocyte lipid deposition in perinatal dairy cows,this experiment proposes the scientific hypothesis that CGA inhibits ERS to alleviate hepatocyte lipid deposition induced by high FAs in calves.Primary calf hepatocytes were isolated and cultured in vitro and stimulated and grouped,and cells stimulated with FAs were divided into four groups,which were the CON group,FAs group,CGA group,and the CGA+FAs group.TG-stimulated cells were also divided into four groups,which were the CON group,TG group,CGA group,and the CGA+TG group.The alleviating effect of CGA on TG-induced ERS could verify its pathway affecting lipid deposition in calf hepatocytes.After grouping,molecular biology and cell biology techniques were used to detect ERS indicators(ATF6,PERK,IREI,CHOP,GRP78),lipid synthesis indicators(FAS,SREBP,ACC1)and lipid droplet morphology distribution in each group.The results showed that after stimulation with FAs and TG,the lipid droplet distribution in calf hepatocytes increased,and the protein expression levels of ERS indicators(ATF6,PERK,IREI,CHOP,GRP78)and lipid synthesis indicators(FAS,SREBP,ACC1)increased.After addition of CGA,the lipid droplet distribution,ERS indicators(ATF6,PERK,IREI,CHOP,GRP78)and lipid synthesis indicators(FAS,SREBP,ACC1)in the two stimulation groups were reduced compared with those stimulated.In conclusion,CGA can alleviate the lipid deposition caused by FAs,and CGA should provoke an alleviating effect on TG-induced ER,so CGA reduces hepatic lipid deposition in calf hepatocytes by alleviating ER stress caused by high FAs.
作者
尹钰锋
李铭
张伟
何雨欣
杨威
王桂华
余丽芸
张冰冰
徐闯
YIN Yufeng;LI Ming;ZHANG Wei;HE Yuxin;YANG Wei;WANG Guihua;YU Liyun;ZHANG Bingbing;XU Chuang(College of Animal Science and Veterinary Medicine,Heilongjiang Bayi Agricultural University,Daqing,Heilongjiang 163319,China;College of Life Science and Technology,Heilongjiang Bayi Agricultural University,Daqing,Heilongjiang 163319,China)
出处
《中国兽医学报》
CAS
CSCD
北大核心
2023年第8期1759-1764,1778,共7页
Chinese Journal of Veterinary Science
基金
黑龙江省自然科学基金优青资助项目(YQ2020C035)
国家自然科学基金联合基金重点资助项目(U20A2062)。
