摘要
目的 研究PI3K/AKT/GSK-3β信号通路在高尿酸血症大鼠尿酸、血糖及脂质代谢中的作用机制。方法 取40只SD大鼠随机分为4组,其中2组为未建模SD大鼠,分别为模型对照组(C组)、LY294002组(L组),剩余2组为建模成功的SD大鼠,分为高尿酸血症组(H组)和高尿酸血症加LY294002干预组(HL组)。C组大鼠经口给予纯水(10 mL/kg体重)和腹腔注射0.2%二甲基亚砜溶液(6 mL/kg体重);L组大鼠给予相同剂量的纯水和腹腔注射LY294002溶液(6 mL/kg体重);H组大鼠经口给予腺嘌呤(100 mg/kg体重)和乙胺丁醇(250 mg/kg体重);HL组大鼠在给予相同剂量的腺嘌呤和乙胺丁醇的同时腹腔注射LY294002溶液(6 mL/kg体重)。各组持续干预15 d,每3天从眼眶静脉丛采集血样,测定血尿酸(Uric Acid, UA)、肌酐(Serum Creatinine, Scr)、尿素氮(Blood Urea Nitrogen, BUN)、血糖(Glucose, GLU)、甘油三酯(Triglyceride, TG)、胆固醇(Total Cholesterol, TC)。第15天麻醉后采集各组大鼠肾脏组织,采用荧光定量PCR法和Western blotting法检测大鼠肾脏组织中PI3K/AKT/GSK-3β的表达水平。结果 4组C组、L组、H组和HL组大鼠实验前血清尿酸、肌酐、尿素氮、血糖、甘油三酯和胆固醇水平差异无统计学意义(P<0.05)。H组和HL组大鼠UA、Scr、BUN水平在实验后均高于C组和L组,与C组第6天比较,H组和HL组UA水平降低,第9天至第15天UA水平升高,差异有统计学意义(P<0.05)。L组、H组和HL组GLU水平较C组升高,其中L组在第12天,GLU水平高于C组,H组和HL组在实验第6天,GLU水平高于L组,差异有统计学意义(P均<0.05)。H组和HL组TC、TG水平较H组和L组升高,其中HL组在实验第9天,TG水平高于H组,差异有统计学意义(P<0.05)。荧光定量PCR检测结果显示,与C组比较,H组和HL组PI3K、AKT、GSK-3β增大,L组AKT减小,差异有统计学意义(P均<0.05)。与L组比较,H组和HL组PI3K、AKTGSK-3β增大,差异有统计学意义(P均<0.05)。与H组比较,HL组PI3K、AKT和GSK-3β增大,差异有统计学意义(P均<0.05)。Western-Blot结果显示,与C组比较,H组和HL组PI3K、AKT、GSK-3β增大,差异有统计学意义(P均<0.05)。与L组比较,H组和HL组PI3K、AKTGSK-3β增大,差异有统计学意义(P均<0.05)。与H组比较,HL组PI3K、AKT和GSK-3β增大,差异有统计学意义(P均<0.05)。结论 PI3K/AKT/GSK-3β信号通路与高尿酸血症之间存在联系,提示该通路对治疗和预防高尿酸血症具有重要意义。
Objective:To investigate the role of PI3K/AKT/GSK-3βsignaling pathway in uric acid,glucose and lipid metabolism in rats with hyperuricemia(HUA).Methods:Forty SD rats were randomly divided into 4 groups,of which[HJ15x]2 groups were unmodeled SD rats,namely model control group(C group)and LY294002 group(L group);the remaining 2 groups were modeled successfully SD rats,which were divided into hyperuricemia group(H group)and hyperuricemia plus LY294002 intervention group(HL group).The rats in the C group were given distilled water(10 mL/kg)orally and intraperitoneally injected with 0.2%DMSO solution(6 mL/kg)daily.The rats in the L group received the same dose of distilled water and intraperitoneally injected LY294002 solution(6 mL/kg).The rats in the H group were administered adenine(100 mg/kg)and ethambutol(250 mg/kg)orally daily.The rats in the HL group were given the same dose[JP2]of adenine and ethambutol along with daily intraperitoneal injection of LY294002 solution(6 mL/kg).The intervention lasted for 15 days,during which blood samples were collected from the orbital venous plexus every 3 days to measure serum uric acid(UA),serum creatinine(Scr),blood urea nitrogen(BUN),glucose(GLU),triglycerides(TG)and total cholesterol(TC).On the 15th day,kidney tissues were collected after anesthesia,and the expression levels of PI3K,AKT and GSK-3βin renal tissues were detected using quantitative real-time PCR and Western blotting.Results:Before the experiment,there were no significant differences in serum UA,Scr,BUN,GLU,TG and TC levels among the C group,L group,H group and HL group.(P<0.05).After the experiment,UA,Scr and BUN levels in the H and HL groups were higher than those in the C and L groups.Compared with the C group on the 6th day,UA levels in the H and HL groups decreased but increased from the 9th to the 15th day,with significant differences(P<0.05).GLU levels in the L,H,and HL groups were higher than those in the C group,with the L group showing higher GLU levels than the C group on the 12th day,and the H and HL groups showing higher GLU levels than the L group on the 6th day(P<0.05).TC and TG levels in the H and HL groups were higher than those in the C and L groups,with TG levels in the HL group being higher than those in the H group on the 9th day(P<0.05).Quantitative real-time PCR results showed that PI3K,AKT and GSK-3βexpression levels were significantly higher in the H and HL groups compared to the C group(P<0.05).In the L group,AKT expression was decreased,while GSK-3βwas increased compared to C group(P<0.05).Compared to L group,PI3K,AKT and GSK-3βlevels were significantly higher in the H and HL groups(P<0.05).Compared to H group,PI3K,AKT and GSK-3βlevels were significantly higher in the HL group(P<0.05).Western blot results also indicated significantly increased the levels of PI3K,AKT and GSK-3βin the H and HL groups compared to the C group(P<0.05),and higher levels in the HL group compared to the H group(P<0.05).Conclusion:[JP2]There may be a potential link between the PI3K/AKT/GSK-3βsignaling pathway and HUA,suggesting the importance of this pathway in future therapeutic and preventive strategies.
作者
巩雪俐
王金洋
杨烨
GONG Xueli;WANG Jinyang;YANG Ye(Department of Pathophysiology,School of Basic Medicine,Xinjiang Medical University,Urumqi 830017,China;Department of Clinical Medicine,Xinjiang Medical University,Urumqi 830017,China;Xinjiang Medical University Second Affiliated Hospital Geriatric Cadres Ward,Urumqi 830011,China)
出处
《新疆医科大学学报》
CAS
2024年第8期1066-1072,共7页
Journal of Xinjiang Medical University
基金
新疆维吾尔自治区杰出青年自然科学基金(2023D01E04)。
