基于动物实验和蛋白组学技术探讨猫须草降尿酸的作用机制OA
Mechanistic Exploration of the Uric Acid-Lowering Effects of Clerodendranthus Spicatus Based on Animal Experiments and Proteomics
目的:基于蛋白组学技术和动物实验探讨猫须草降尿酸(UA)的作用机制.方法:健康SPF大鼠40只,随机分为正常组、模型组、阳性药苯溴马隆组和猫须草高、低剂量组.正常组给予蒸馏水,其余各组大鼠给予10%果糖水建立高尿酸血症(HUA)模型,造模同时,阳性药苯澳马隆组大鼠灌胃苯溴马隆片10 mg(kg·d),猫须草高、低剂量组大鼠灌胃10.5 g/(kg·d)猫须草,均给药4周.检测各组大鼠给药第1、2、3、4周血清UA、尿素氮(BUN)、肌酐(CRE);苏木素-伊红(HE)染色观察肾脏形态;实时荧光定量PCR(RT-PCR)检测肾脏三磷酸腺苷结合转运蛋白G2(ABCG2)、葡萄糖转运体9(GLUT9)和尿酸盐转运体(URAT1)基因的表达;蛋白组学检测肾脏差异蛋白.结果:与正常组比较,模型组大鼠血清UA水平显著升高(P<0.01).与模型组比较,给药第1、2、3、4周,阳性药苯溴马隆组和猫须草高剂量组大鼠血清UA含量显著降低(P<0.01,P<0.05);给药第3周和第4周,猫须草低剂量组血清UA含量显著降低(P<0.05),而在其他给药时间点无显著变化(P>0.05).实验期间,各组大鼠血清BNU水平无显著变化(P>0.05).与CON组比较,实验第3周和第4周,模型组大鼠血清CRE显著升高(P<0.05).与MOD组比较,实验第4周,猫须草高剂量组大鼠血清CRE显著降低(P<0.05),而其余各组无明显变化(P>0.05).HE染色显示,正常组大鼠肾组织肾小球、肾小管正常.与正常组比较,模型组大鼠肾小管上皮细胞胞质淡染,肾小管明显扩张,部分细胞核排列紊乱.与模型组比较,阳性药苯溴马隆组肾小管扩张明显改善,猫须草高、低剂量组大鼠肾小球和肾小管形态结构均明显改善.与正常组比较,模型组肾脏组织中的GLUT9和URAT1显著升高,ABCG2显著降低(P<0.01).与模型组比较,不同剂量的猫须草均可显著降低大鼠肾脏组织内的URAT1基因的表达,升高ABCG2基因的表达(P<0.01,P<0.05),猫须草高剂量组GLUT9基因表达显著降低(P<0.01).蛋白组学结果显示,猫须草高剂量组和模型组共有1 936个蛋白发生变化,其中1 082个蛋白表达上调,854个蛋白表达下调,涉及调控p53信号通路、自噬和mTOR信号通路等.结论:猫须草发挥降UA的作用机制可能与改善肾功能,调节ABCG2、GLUT9和URAT1的基因表达,抑制p53 信号通路、自噬和mTOR信号通路有关.
Objective:To investigate the mechanism by which Clerodendranthus Spicatus lowers uric acid(UA)levels using proteomics and animal experiments.Methods:Forty healthy SPF rats were randomly divided into five groups:Normal group,Model group,Positive drug Benzbromarone group,and High-and Low-dose Clerodendranthus Spicatus groups.The Normal group received distilled water,while the other groups were administered 10%fructose water to establish a hyperuricemia(HUA)model.Simultaneously,the Positive drug group received Benzbromarone(10 mg/kg·d),and the Clerodendranthus Spicatus High-and Low-dose groups received Clerodendranthus Spicatus(10.5 g/kg·d)for 4 weeks.Serum UA,blood urea nitrogen(BUN),and creatinine(CRE)levels were measured at 1,2,3,and 4 weeks.Kidney morphology was observed using hematoxylin-eosin(HE)staining.Real-time quantitative PCR(RT-PCR)was employed to assess the expression of ATP-binding cassette sub-family G member 2(ABCG2),glucose transporter 9(GLUT9),and urate transporter 1(URAT1)genes in the kidneys.Proteomics was used to identify differential proteins in the kidneys.Results:Compared to the Normal group,the Model group exhibited significantly elevated serum UA levels(P<0.01).Compared to the Model group,the Positive drug and High-dose Clerodendranthus Spicatus groups had significantly reduced serum UA levels at weeks 1,2,3,and 4(P<0.01,P<0.05).The Low-dose Clerodendranthus Spicatus group showed significant reductions in serum UA at weeks 3 and 4(P<0.05),but not at other time points(P>0.05).No significant changes in serum BUN levels were observed across groups(P>0.05).Compared to the Control group,the Model group had significantly elevated serum CRE levels at weeks 3 and 4(P<0.05).Compared to the Model group,the High-dose Clerodendranthus Spicatus group had significantly reduced serum CRE levels at week 4(P<0.05),while other groups showed no significant changes(P>0.05).HE staining revealed normal kidney glomeruli and tubules in the Normal group,but the Model group showed light staining in renal tubular epithelial cells,severe tubular dilation,and disordered nuclear arrangement.The Positive drug and High-and Low-dose Clerodendranthus Spicatus groups exhibited significant improvements in kidney structure.Compared to the Normal group,the Model group had significantly increased GLUT9 and URAT1 levels and decreased ABCG2 levels(P<0.01).All doses of Clerodendranthus Spicatus significantly reduced URAT1 expression and increased ABCG2 expression(P<0.01,P<0.05),with the High-dose Clerodendranthus Spicatus group showing a significant reduction in GLUT9 expression(P<0.01).Proteomics identified 1,936 proteins with altered expression in the High-dose Clerodendranthus Spicatus and Model groups,with 1,082 proteins upregulated and 854 downregulated,involving pathways related to p53 signaling,autophagy,and mTOR signaling.Conclusion:The uric acid-lowering effects of Clerodendranthus Spicatus may be related to its ability to improve kidney function,regulate the expression of ABCG2,GLUT9,and URAT1 genes,and inhibit the p53 signaling pathway,autophagy,and mTOR signaling.
吴梦娟;朱青春;贾丽阳;朱春胜
郑州大学第一附属医院,河南 郑州 450052河南省中医院,河南 郑州 450046
猫须草高尿酸血症尿酸转运体蛋白组mTOR信号通路
Clerodendranthus SpicatusHyperuricemiaUric acid transportersProteomicsmTOR signaling pathway
《中医药信息》 2024 (010)
1-8 / 8
国家自然科学基金项目(82204701)
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