荆防颗粒对氧嗪酸钾诱导小鼠高尿酸肾病的防治作用机制探讨  

Preventive and therapeutic mechanism of Jingfang granules on hyperuricemia nephropathy induced by potassium oxyzinate in mice

作　　者：尉雅洁 刘明飞 周诗喆 项海鑫 孙成宏 陈颖[1] YU Yajie;LIU Mingfei;ZHOU Shizhe;XIANG Haixin;SUN Chenghong;CHEN Ying(Department of Endocrinology and Metabolism,The Affiliated Hospital of Qingdao University,Qingdao 266003,China;不详)

机构地区：[1]青岛大学附属医院内分泌与代谢性疾病科,山东青岛266003 [2]中国海洋大学医学院海洋药物重点实验室 [3]鲁南制药集团股份有限公司中药制药共性技术国家重点实验室

出　　处：《山东医药》2023年第15期1-5,共5页Shandong Medical Journal

基　　金：国家自然科学基金青年科学基金项目(81600601);山东省自然科学基金面上项目(ZR2021MH363)。

摘　　要：目的探讨荆防颗粒对氧嗪酸钾诱导小鼠糖尿病肾病的防治作用机制。方法采用靶点“钩钓”方法制备荆防颗粒有效成分键合的磁性微球,将携带荆防颗粒有效成分的磁性微球与小鼠的正常肾组织裂解液共同孵育,捕获荆防颗粒潜在结合靶点蛋白;再采用高分辨质谱筛选荆防颗粒有效成分特异性结合的靶点蛋白。采用京都基因与基因组百科全书(KEGG)分析靶点蛋白富集的信号通路。将36只ICR雄性小鼠按体重随机分为荆防颗粒低剂量组、荆防颗粒中剂量组、荆防颗粒高剂量组、模型组、别嘌呤醇(AL)组、对照组,每组6只。除腹腔注射250 mg/kg氧嗪酸钾溶液外,荆防颗粒低、中、高剂量组分别灌胃荆防颗粒0.5、1、2 g/kg;模型组灌胃相应体积纯化水;AL组灌胃25 mg/(kg·d)AL溶液;对照组灌胃相应体积纯化水;均1次/d,连续用药4周。末次给药后1 h,测定血清尿酸(UA)、尿素氮(BUN)和肌酐(Cr),HE染色观察肾组织病理变化,Western blotting法检测靶点蛋白相关信号通路的主要指标(氧化应激指标MDA、SOD、GSH、Nrf2、HO-1、NQO1,黏膜屏障功能蛋白Occludin、ZO-1)。结果从肾组织中鉴定到荆防颗粒176个靶点蛋白,靶点蛋白的功能可能与过氧化物酶体、谷胱甘肽代谢、紧密连接和黏着连接等通路相关。与对照组相比,模型组小鼠血清UA、BUN和Cr水平升高(P均<0.05);与模型组相比,荆防颗粒和AL处理组血清UA、BUN和Cr水平降低,且荆防颗粒组呈现明显剂量依赖性(P均<0.05)。中、高剂量荆防颗粒组小鼠肾皮质趋于正常结构,肾小管形态改善、炎症明显减轻,尤其是高剂量组;AL组小鼠间质内有少量炎症细胞浸润,较模型组有所改善。与对照组相比,模型组小鼠肾组织中MDA含量升高,SOD、GSH活性降低(P均<0.05);Nrf2、HO-1、NQO1蛋白相对表达量降低;Occludin、ZO-1蛋白相对表达量降低。与模型组相比,荆防颗粒各剂量组及AL组MDAObjective To explore the preventive and therapeutic mechanism of Jingfang granules(JF)on hyperuri⁃cemia nephropathy induced by potassium oxyzinate(PO)in mice.Methods“Target fishing”strategy was used to pre⁃pare magnetic microspheres bound to the effective ingredients of JF.The magnetic microspheres with effective ingredients of JF incubated with normal renal tissue lysates of mice,and the potential binding target proteins were directly captured from the tissue lysates.High-resolution mass spectrometry was used to screen the target proteins that specifically bound to the effective ingredients of JF.The Kyoto Encyclopedia of Genes and Genomes(KEGG)was used to analyze the signaling pathways enriched in target proteins.Thirty-six male ICR mice were randomly divided into the low-dose JF group,medium-dose JF group,high-dose JF group,model group,allopurinol(AL)group,and control group according to body weight,with 6 mice in each group.In addition to intraperitoneal injection of 250 mg/kg PO solution,mice in the low-,medium-,and high-dose JF groups were gavaged with 0.5,1,and 2 g/kg JF,respectively;mice in the model group were gavaged with corresponding volume of purified water,mice in the AL group were gavaged with 25 mg/(kg·d)AL solution,and mice in the control group were gavaged with corresponding volume of purified water.All were administered once a day for 4 consecutive weeks.One hour after the last administration,blood was taken to measure serum uric acid(UA),urea nitrogen(BUN),and creatinine(Cr).Renal tissue was observed by H&E staining.Main indicators of oxidative stress,such as malonaldehyde(MDA),superoxide dismutase(SOD),and glutathione peroxidase(GSH)were detected by ELI⁃SA.Main indicators of target protein-related signaling pathways(oxidative stress proteins including Nrf2,HO-1 and NQO1,and mucosal barrier functional proteins including Occludin and ZO-1)were detected by Western blotting.Re⁃sults Totally 176 specific binding proteins were identified from the renal tissues,and the function of target

关 键 词：荆防颗粒 高尿酸肾病 氧化应激 黏膜屏障 

分 类 号：R285.5[医药卫生—中药学]