前列腺素家族中的PGB2、15-keto-PGE2、8-iso-PGF2α在非酒精性脂肪性肝病中的作用  

Roles of prostaglandin B2,15⁃keto⁃prostaglandin E2,and 8⁃isoprostane F2αin non⁃alcoholic fatty liver disease

作　　者：高毅男 王培君 叶棣文 郭泽郡 逯素梅[1] GAO Yinan;WANG Peijun;YE Diwen;GUO Zejun;LU Sumei(Department of Laboratory Medicine,the First Affiliated Hospital of Shandong First Medical University&Shandong Provincial Qianfoshan Hospital,Shandong Medicine and Health Key Laboratory of La-boratory Medicine,Jinan 250014,Shandong;Department of Laboratory Medicine,Jinan Central Hospital,Jinan 250013,Shan-dong;School of Medical Laboratory,Weifang Medical College,Weifang 261053,Shandong,China)

机构地区：[1]山东第一医科大学第一附属医院/山东省千佛山医院检验科,山东省医药卫生临床检验诊断学重点试验室,济南250014 [2]济南市中心医院检验科,济南250013 [3]潍坊医学院医学检验学院,山东潍坊261053

出　　处：《临床检验杂志》2024年第7期535-541,共7页Chinese Journal of Clinical Laboratory Science

基　　金：山东第一医科大学青年科学基金培育资助计划(202201⁃083);山东省自然科学基金资助项目(ZR2023MH031,ZR2021MH187)。

摘　　要：目的探讨前列腺素家族(prostaglandins,PGs)成员对非酒精性脂肪肝(non-alcoholic fatty liver disease,NAFLD)的影响。方法以人肝癌细胞系HepG2细胞为研究对象。试验设为对照组(Ctrl)、脂肪变组(FFA)、前列腺素B2(PGB2、10μg/mL)处理组、15-酮基-前列腺素E2(15-keto-PGE2、10μg/mL)处理组、8-异前列腺素F2a(8-iso-PGF2α、10μg/mL)处理组。采用噻唑蓝(MTT)试验检测细胞活性,油红O染色检测脂质沉积,实时荧光定量PCR(qRT-PCR)检测炎性因子基因的表达情况,Western blot检测磷酸化胰岛素受体底物(p-IRS)蛋白的表达水平。另取15只SPF级雄性C57BL/6J小鼠,随机分为基础组(CD组,n=5,10%低脂饲料喂养16周)、高脂组(HFD组,n=5,60%高脂饲料喂养16周,建模NAFLD)、PGB2组(n=5,60%高脂饲料喂养16周后,每天给予尾静脉注射PGB220μg/kg,持续2周)。采用小鼠葡萄糖耐量试验(IPGTT)检测小鼠的糖耐量水平,HE染色验证小鼠肝脏脂肪变程度。结果油红O染色结果显示,PGs对NAFLD的脂质沉积没有显著影响,但PGs能够缓解NAFLD伴随产生的炎症。qRT-PCR结果显示,FFA组IL-1β水平[(2.274±0.550)倍]与对照组相比显著升高(P=0.0028),而在50μg/mL PGB2、10μg/mL 15-keto-PGE2和10μg/mL 8-iso-PGF2α的作用下,IL-1β分别降低至[(0.720±0.036)倍,P=0.0031、(0.857±0.225)倍,P=0.0064和(1.767±0.725)倍,P=0.0297]。Western blot结果显示,经过PGs处理,p-IRS蛋白的表达水平增加。CD组小鼠体重为(28.560±2.028)g,HFD组小鼠体重升高至(49.300±0.667)g,而PGB2组显著降低至(40.840±4.043)g,各组间差异有统计学意义(P=0.0017);此外,PGB2组葡萄糖耐量结果优于HFD组。HE染色结果显示,与HFD组比较,PGB2组肝脏脂肪变的程度降低。结论PGs中的PGB2、15-keto-PGE2、8-iso-PGF2α可通过缓解IL-1β介导的炎症,上调p-IRS表达,促进胰岛素信号传导,减轻胰岛素抵抗,缓解NAFLD的发生、发展。Objective To investigate the effect of prostaglandin family(PGs)on non-alcoholic fatty liver disease(NAFLD).Methods HepG2 cells,a human hepatocellular carcinoma cell line,were used as the research subject.The experiment was set up as a control group(Ctrl),fatty change group(FFA),prostaglandin B2(PGB2,10μg/mL)treatment group,15-keto-prostaglandin E2(15-keto-PGE2,10μg/mL)treatment group,and 8-iso-prostaglandin F2a(8-iso-PGF2α,10μg/mL)treatment group.Cell activity was determined by the thiazolyl blue(MTT)assay and lipid deposition was detected by the oil red O staining.The expression levels of inflammatory factors and phosphorylated insulin receptor substrates(p-IRS)were determined by real-time fluorescence quantitative PCR(qRT-PCR)and Western blot,respectively.In addition,15 SPF-grade male C57BL/6J mice were randomly divided into a basic group(CD group,n=5,fed with 10%low-fat forage for 16 weeks),high-fat group(HFD group,n=5,fed with 60%high-fat forage for 16 weeks to model NAFLD),and PGB2 group(n=5,given 20μg/kg PGB2 daily by tail vein injection for 2 weeks after 16 weeks of 60%high-fat diet feeding).The glucose tolerance level of the mice was detected by the intraperitoneal glucose tolerance test(IPGTT)and the degree of hepatic steatosis was determined by HE staining.Results Oil red O staining showed that PGs had no sig-nificant effect on the lipid deposition of NAFLD,but PGs were able to alleviate the inflammation associated with NAFLD.qRT-PCR re-sults showed that compared with the Ctrl group,the levels of IL-1βin the FFA group increased by 2.274±0.550 times(P=0.0028),while under the action of 50μg/mL PGB2,10μg/mL 15-keto-PGE2 and 10μg/mL 8-iso-PGF2α,the levels of IL-1βdecreased to 0.720±0.036 times(P=0.0031),0.857±0.225 times(P=0.0064),and 1.767±0.725 times(P=0.0297),respectively.Western blot results showed that after PGs treatment,the expression level of p-IRS protein was increased.The body weights of mice in the CD group,HFD group and PGB2 group were(28.560±2.028)g,(49.300±0.667)g,and(40.840�

关 键 词：前列腺素 非酒精性脂肪肝 炎症 胰岛素抵抗 磷酸化胰岛素受体底物 

分 类 号：R446[医药卫生—诊断学] R589[医药卫生—临床医学]