高脂诱导下雄激素缺乏小型猪肾脏脂质沉积的关键基因表达分析  

Analysis of key genes regulating renal lipid accumulation in testosteronedeficient miniature pigs fed with high-fat diet

作　　者：范莹盈 任裕杰 凌云 吕东颖 蔡兆伟 FAN Yingying;REN Yujie;LING Yun;LÜDongying;CAI Zhaowei(School of Pharmaceutical Sciences,Zhejiang Chinese Medical University,Hangzhou 310053,China;Laboratory Animal Research Center,Academy of Chinese Medical Science,Zhejiang Chinese Medical University,Hangzhou 310053,China)

机构地区：[1]浙江中医药大学药学院,浙江杭州310053 [2]浙江中医药大学中医药科学院动物实验研究中心,浙江杭州310053

出　　处：《中国病理生理杂志》2023年第2期305-313,共9页Chinese Journal of Pathophysiology

基　　金：浙江省基础公益研究计划项目(No.LGD22C040008);浙江中医药大学自然科学青年探索项目(No.2021JKZKTS008A)。

摘　　要：目的:探讨高脂诱导条件下雄激素缺乏小型猪肾脏脂质沉积及关键基因表达的变化。方法:将雄性五指山小型猪随机分为3组,即不去势(IM)组、去势(CM)组和去势+睾酮(CMT)组,每组6只动物,均饲喂高脂饮食。12周后检测血清肾功能指标,测定肾脏甘油三酯(TG)和总胆固醇(TC)含量;进行肾脏苏木精-伊红(H&E)和油红O染色,观察其脂质沉积和组织病理学变化。利用转录组测序分析肾脏组织表达谱差异,并用RT-qPCR和Western blot方法验证参与TG合成、胆汁酸代谢和雌激素合成相关差异表达基因。结果:CM小型猪肾脏重量明显低于IM和CMT小型猪(P<0.05);与IM组和CMT组小型猪相比,CM小型猪血尿素氮含量显著升高(P<0.05),但血清肌酐和总蛋白水平没有显著变化;CM组小型猪肾脏内出现大量脂滴,且TG含量显著高于IM和CMT小型猪(P<0.05);与IM组和CMT组小型猪相比,CM组小型猪肾脏TG合成基因包括固醇调节元件结合转录因子1(SREBF1)、糖类应答元件结合蛋白(ChREBP/MLXIPL)和硬脂酰辅酶A去饱和酶(SCD)等表达升高,而胆汁酸代谢和雌激素合成基因包括法尼酯X受体(FXR/NR1H4)和雌激素受体1(ESR1)表达下调;睾酮处理能够逆转去势小型猪肾脏内脂质沉积及相关基因表达变化。结论:ESR1和NR1H4可能通过影响SREBF1脂质合成途径参与高脂诱导的雄激素缺乏小型猪肾脏脂质沉积过程。这为老年男性慢性肾脏疾病防治提供了新思路。AIM: To investigate the effect of testosterone deficiency on renal lipid deposition and related gene expression in miniature pigs fed with high-fat diet. METHODS: Male Wuzhishan miniature pigs were randomly divided into 3 groups: intact male pigs(IM), castrated male pigs(CM) and castrated male pigs with testosterone replacement(CMT). Six pigs in each group were fed with high-fat diet for 12 weeks. Serum renal function parameters and renal triglyceride(TG) and total cholesterol(TC) levels were detected. Oil red O and HE staining was performed to observe lipid deposition and histopathological changes in the kidney. RNA-Seq technology was used to investigate the differential expression of transcriptome in the kidney. RT-qPCR and Western blot were used to verify the differentially expressed genes involved in TG synthesis, bile acid metabolism and estrogen synthesis. RESULTS: The kidney weight was significantly lower in CM group than IM group and CMT group(P<0. 05). The content of blood urea nitrogen in CM group was increased compared with IM group and CMT group(P<0. 05). However, serum creatinine and total protein levels showed no significant difference. A large number of lipid droplets and significantly increased TG content in the kidney were observed in CM group compared with IM and CMT groups(P<0. 05). Compared with IM group and CMT group, the expression of TG synthesis genes including sterol regulatory element binding transcription factor 1(SREBF1), carbohydrate-responsive element-binding protein(ChREBP/MLXIPL) and stearoyl-coenzyme A desaturase(SCD) was significantly increased in the kidneys of CM, while the expression of genes related to bile acid metabolism and estrogen synthesis including farnesoid X receptor(FXR/NR1H4) and estrogen receptor 1(ESR1) was significantly decreased. Testosterone replacement reversed the changes of lipid deposition and related gene expression in the kidneys of CM. CONCLUSION: The ESR1 and NR1H4 genes may be involved in the process of lipid deposition in the kidneys of CM fed with h

关 键 词：肾脏 脂质沉积 雄激素缺乏 小型猪 高脂饮食 

分 类 号：R692[医药卫生—泌尿科学] R363.2[医药卫生—外科学]