植物提取物复合制剂缓解奶牛乳腺上皮细胞炎症反应的分子机制  

Molecular Mechanisms of Plant Extracts Compound Preparation in Relieving Inflammatory Responses for Bovine Mammary Epithelial Cells

作　　者：张腾龙[1] 郭晨阳 宋洁 敖长金[1] 王丽芳[2] ZHANG Tenglong;GUO Chenyang;SONG Jie;AO Changjin;WANG Lifang(College of Animal Science,Inner Mongolia Agricultural University,Hohhot 010010,China;Inner Mongolia Academy of Agricultural&Animal Husbandry Sciences,Hohhot 010031,China)

机构地区：[1]内蒙古农业大学动物科学学院,呼和浩特010010 [2]内蒙古农牧业科学院,呼和浩特010031

出　　处：《动物营养学报》2023年第6期3955-3968,共14页CHINESE JOURNAL OF ANIMAL NUTRITION

基　　金：国家自然科学基金项目(31860663);内蒙古自治区科技计划项目(2021GG0029);内蒙古自然科学基金项目(2021LHMS03014);内蒙古农牧业创新基金项目(2020CXJJM12);奶牛疾病绿色防控技术应用示范(2022TG16)。

摘　　要：本试验旨在研究植物提取物复合制剂对脂多糖(LPS)诱导的奶牛乳腺上皮细胞(BMEC)炎症模型免疫、抗氧化功能和转录组的影响,以揭示植物提取物复合制剂缓解BMEC炎症反应的分子机制。试验选用第3代贴壁生长BMEC,随机分为5组,每组8个重复,每个重复1个培养孔。对照组BMEC不添加LPS和植物提取物复合制剂,采用基础培养基培养10 h;LPS组BMEC添加10μg/mL LPS作用6 h后,采用基础培养基培养4 h;试验组BMEC加入10μg/mL LPS作用6 h后,分别加入30(WELPS30组)、50(WELPS50组)及70μg/mL(WELPS70组)的植物提取物复合制剂,采用基础培养基培养4 h。测定各组免疫及抗氧化相关指标,并选择LPS组和WELPS30组进行转录组测序及分析。结果表明:1)LPS组BMEC相对增殖率显著低于对照组(P<0.05),WELPS30组BMEC相对增殖率显著高于LPS组(P<0.05),且WELPS30组、WELPS50组和WELPS70组BMEC中肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)及白细胞介素-8(IL-8)含量显著低于LPS组(P<0.05)。2)LPS组BMEC中丙二醛(MDA)含量和诱导性一氧化氮合酶(iNOS)活性显著高于对照组(P<0.05),WELPS30组、WELPS50组和WELPS70组BMEC中MDA含量和iNOS活性显著低于LPS组(P<0.05),WELPS30组BMEC中总超氧化物歧化酶(T-SOD)、过氧化氢酶(CAT)和谷胱甘肽过氧化物酶(GSH-Px)活性及总抗氧化能力(T-AOC)显著高于LPS组(P<0.05)。3)转录组测序结果表明,共有369个差异表达基因。GO富集分析显示,差异表达基因主要参与调节白细胞介素-17(IL-17)生成、白细胞介导的细胞毒素作用、T细胞增殖等生物学过程。KEGG富集分析显示,差异表达基因主要集中在以NOD样受体(NLR)信号通路和IL-17信号通路为主的免疫相关信号通路。4)以介数中心性为指标进行网络分析,发现Toll样受体2(TLR2)、趋化因子10(CXCL10)和集落刺激因子2(CSF2)等10个基因在植物提取物复合制剂调控的差异表达免疫基因中起到�The aim of this study was to investigate the effects of plant extracts compound preparation on the im⁃mune,antioxidant function and transcriptome of lipopolysaccharide(LPS)⁃induced bovine mammary epithelial cells(BMEC),and to reveal the molecular mechanism of plant extracts compound preparation in relieving in⁃flammatory response for BMEC.The third⁃generation attachment cultured BMEC were randomly divided into 5 groups with 8 replicates per group and 1 culture pore per replicate.BMEC in control group were cultured in a basal medium without plant extracts compound preparation and LPS for 10 h;BMEC in LPS group were ex⁃posed in LPS with 10μg/mL for 6 h,and then cultured in a basal medium for 4 h;BMEC in experimental groups were exposed in LPS with 10μg/mL for 6 h,then supplemented with 30(WELPS30 group),50(WELPS50 group)and 70μg/mL(WELPS70 group)plant extracts compound preparation,respectively,and then cultured in basal mediums for 4 h.The immune and antioxidant related indexes were detected,LPS group and WELPS30 group were selected for transcriptome sequencing and analysis.The results showed as fol⁃lows:1)the BMEC relative proliferation rate of LPS group was significantly lower than that of control group(P<0.05),the BMEC relative proliferation rate of WELPS30 group,WELPS50 group and WELPS70 group was significantly higher than that of LPS group(P<0.05),and the contents of tumor necrosis factor⁃α(TNF⁃α),interleukin⁃1β(IL⁃1β),interleukin⁃6(IL⁃6)and interleukin⁃8(IL⁃8)in BMEC of WELPS30 group,WELPS50 group and WELPS70 group were significantly lower than those of LPS group(P<0.05).2)The content of malondialdehyde(MDA)and activity of inducible nitric oxide synthase(iNOS)in BMEC of LPS group were significantly higher than those in control group(P<0.05),the content of MDA and activity of iN⁃OS in BMEC of WELPS30 group,WELPS50 group and WELPS70 group were significantly lower than those of LPS group(P<0.05),the activities of total superoxide dismutase(T⁃SOD),catalase(CAT)and glutathi�

关 键 词：植物提取物复合制剂 奶牛乳腺上皮细胞 炎症反应 转录组 

分 类 号：S823[农业科学—畜牧学]