作者：李桐，朱天麒，方少婷，欧阳军艳，向惠芳，陈赛珍，王耀霆，姚雨涵，黄春霞

单位：

410219湖南省长沙市，长沙医学院

Units：

Changsha Medical College, Changsha 410219, China

关键词：

高尿酸血症；葛根；异黄酮类；炎症；应激，生理学

Keywords：

Hyperuricemia; Puerariae lobamle radix; Isoflavones; Inflammation; Stress, physiological

CLC：

R 589.9

DOI：

10.12114/j.issn.1008-5971.2022.00.040

Funds：

国家级大学生创新创业训练计划项目（S202010823014）；2020年度湖南省大学生创新创业训练计划项目（3897，1087）

摘要：

背景 人类因缺乏尿酸酶基因而无法代谢不溶性尿酸（UA），而高嘌呤饮食、代谢疾病均会导致 UA升高，病情严重者可诱发肾功能不全等疾病。黄酮类化合物对多种酶有抑制作用，异黄酮是中药葛根的主要功 效成分，其已被证实具有明确的降糖、降脂作用。目的　探讨葛根异黄酮对高尿酸血症（HUA）小鼠炎症、应激反 应的影响。方法　2020年12月至2021年6月，采用随机数字表法将48只SPF级昆明小鼠分为正常组、模型组、条件对 照组、实验组，各12只。正常组小鼠每天灌胃0.5%羧甲基纤维素钠；其余三组小鼠每天灌胃等体积腺嘌呤和盐酸乙 胺丁醇混合液以制备HUA模型，共计30 d。造模成功后正常组小鼠继续用混合饲料饲养，不予其他药物；模型组小 鼠继续灌胃0.5%羧甲基纤维素钠，不予其他药物；条件对照组小鼠继续灌胃0.5%羧甲基纤维素钠，同时灌胃非布司 溶液；实验组小鼠继续灌胃0.5%羧甲基纤维素钠，同时灌胃葛根异黄酮；四组小鼠干预时间均为30 d。比较四组小 鼠干预前后血清UA、炎症指标〔包括P物质（SP）、红细胞沉降率（ESR）、C反应蛋白（CRP）、肿瘤坏死因子 α（TNF-α）、白介素6（IL-6）〕、应激反应指标〔包括超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GSHPx）、丙二醛（MDA）、一氧化氮（NO）、一氧化氮合酶（NOS）〕。观察四组小鼠主动脉弓内皮细胞病理学特征。 结果　干预后，模型组、条件对照组、实验组小鼠血清UA高于正常组，条件对照组、实验组小鼠血清UA低于模型组 （P＜0.05）；模型组、条件对照组、实验组小鼠干预后血清UA分别高于本组干预前（P＜0.05）。干预后，模型组、 条件对照组、实验组小鼠血清SP、ESR、CRP、TNF-α高于正常组，条件对照组、实验组小鼠血清SP、ESR、CRP、 TNF-α低于模型组，实验组小鼠血清SP、ESR、CRP低于条件对照组（P＜0.05）；模型组、条件对照组、实验组小鼠 干预后血清SP、ESR、CRP、TNF-α分别高于本组干预前（P＜0.05）。干预后，模型组、条件对照组、实验组小鼠 血清SOD高于正常组，血清GSH-Px低于正常组（P＜0.05）；干预后，模型组、条件对照组小鼠血清MDA高于正常组 （P＜0.05）；干预后，条件对照组、实验组小鼠血清SOD、MDA低于模型组，血清GSH-Px高于模型组（P＜0.05）； 干预后，实验组小鼠血清SOD、MDA低于条件对照组，血清GSH-Px高于条件对照组（P＜0.05）。模型组、条件对照 组、实验组小鼠干预后血清SOD、MDA分别高于本组干预前，血清GSH-Px分别低于本组干预前（P＜0.05）。正常组 小鼠主动脉弓内皮细胞排列规则，无明显裂缝。模型组小鼠主动脉弓内皮细胞有明显水样变性，胞质以及细胞核轻微 肿胀，部分区域有淋巴细胞。条件对照组和实验组小鼠主动脉弓部分内皮细胞有双核，实验组小鼠内皮细胞恢复情况 较好，细胞形态比较完整。结论　与非布司溶液相比，葛根异黄酮可更有效地减轻HUA小鼠的病情严重程度，同时减 轻炎症和应激反应，改善心血管功能。

Abstract：

Background Humans cannot metabolize insoluble uric acid (UA) due to the lack of uricase gene, and high purine diet and metabolic diseases can lead to increased UA, and severe cases can induce diseases such as renal insufficiency. Flavonoids have inhibitory effects on a variety of enzymes, isoflavones are the main functional components of traditional Chinese medicine Pueraria, which have been confirmed to have clear hypoglycemic and lipid-lowering effects. Objective To investigate the effect of pueraria isoflavones on inflammation and stress response in hyperuricemia (HUA) mice. Methods From December 2020 to June 2021, 48 SPF Kunming mice were divided into normal group, model group, condition control group, and experimental group by random number table method, with 12 mice in each group. The mice in the normal group were given 0.5% sodium carboxymethyl cellulose by gavage every day; the mice in the other three groups were given an equal volume of a mixture of adenine and ethambutol hydrochloride by gavage every day to make the HUA model, for a total of 30 d. After successful modeling, the mice in the normal group were fed with mixed feed without other drugs; the mice in the model group were continuously gavaged with 0.5% carboxymethyl cellulose sodium without other drugs; the mice in the condition control group were continuously gavaged with 0.5% carboxymethyl cellulose sodium and fibrate solution at the same time; the mice in the experimental group were gavaged with 0.5% carboxymethyl cellulose sodium and pueraria isoflavones at the same time; the intervention time of the four groups was 30 d. Serum UA, inflammatory indexes [including substance P (SP) , erythrocyte sedimentation rate (ESR) , C-reactive protein (CRP) , tumor necrosis factor alpha (TNF-α) , interleukin 6 (IL-6) ] , and stress response indexes [including superoxide dismutase (SOD) , glutathione peroxidase (GSH-Px) , malondialdehyde (MDA) , nitric oxide (NO) , nitric oxide synthase (NOS) ] before and after intervention in four groups were compared. The pathological characteristics of aortic arch endothelial cells in four groups were observed. Results After intervention, the serum UA of the mice in the model group, condition control group and experimental group was higher than that of the normal group, and the serum UA of the mice in the condition control group and experimental group was lower than that of the model group (P < 0.05) ; the serum UA of mice in the model group, condition control group and experimental group after intervention was higher than that before intervention, respectively (P < 0.05) . After intervention, the serum SP, ESR, CRP and TNF-α of the mice in the model group, conditional control group and experimental group were higher than those in the normal group, the serum SP, ESR, CRP and TNF-α of the mice in the condition control group and experimental group were lower than those in the model group, and the serum SP, ESR and CRP of the mice in the experimental group were lower than those in the condition control group (P < 0.05) ; the serum SP, ESR, CRP and TNF-α of mice in the model group, condition control group and experimental group after intervention were higher than those before intervention, respectively (P < 0.05) . After intervention, the serum SOD of the mice in the model group, condition control group and experimental group was higher than that of the normal group, and the serum GSH-Px of the mice was lower than that of the normal group (P < 0.05) ; after intervention, the serum MDA of the mice in the model group and condition control group was higher than that in the normal group (P < 0.05) ; after intervention, the serum SOD and MDA of the mice in the condition control group and experimental group were lower than those in the model group, and serum GSH-Px was higher than that in the model group (P < 0.05) ; after intervention, the serum SOD and MDA of the mice in the experimental group were lower than those in the conditional control group, and the serum GSH-Px was higher than that in the conditional control group (P < 0.05) . The serum SOD and MDA of mice in the model group, condition control group and experimental group after intervention were higher than those before intervention, and serum GSH-Px were lower than those before intervention, respectively (P < 0.05) . The endothelial cells of the aortic arch of mice in the normal group were arranged regularly without obvious cracks. The endothelial cells of the aortic arch of mice in the model group had obvious watery degeneration, the cytoplasm and nucleus were slightly swollen, and there were lymphocytes in some areas. Some aortic arch endothelial cells in the condition control group and experimental group had binucleated cells, and the endothelial cells in the experimental group recovered well and the cell morphology was relatively complete. Conclusion Compared with febux solution, pueraria isoflavones can more effectively reduce disease severity in HUA mice while attenuating inflammatory and stress responses.

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