目的 探究莫諾苷改善果糖飲食誘導(dǎo)小鼠腎損傷的作用及其機(jī)制。方法 將48只雄性C57BL/6小鼠隨機(jī)分為對(duì)照組、模型組、別嘌呤醇（25 mg/kg）組及莫諾苷低、高劑量（25、50 mg/kg）組和莫諾苷（50 mg/kg）＋谷胱甘肽過(guò)氧化酶4（glutathione peroxidase 4，GPX4）抑制劑RSL3（5 mg/kg）組。對(duì)照組小鼠正常飲水，其余各組小鼠均給予30%果糖水飼養(yǎng)，同時(shí)各給藥組小鼠給予相應(yīng)藥物，1次/d，連續(xù)給藥8周。實(shí)驗(yàn)結(jié)束后收集各組小鼠血清、尿液和腎臟樣本，記錄各組小鼠體質(zhì)量、腎臟質(zhì)量，并計(jì)算腎臟指數(shù)；檢測(cè)各組小鼠血清中肌酐（creatinine，Cre）、尿素氮（blood urea nitrogen，BUN）和尿酸（uric acid，UA）的含量；檢測(cè)各組小鼠尿液中尿蛋白的含量；同時(shí)檢測(cè)各組小鼠腎臟中活性氧（reactive oxygen species，ROS）、丙二醛（malondialdehyde，MDA）、總超氧化物歧化酶（total-superoxide dismutase，T-SOD）、脂質(zhì)過(guò)氧化物（lipid hydroperoxide，LPO）、總鐵和亞鐵離子（Fe²⁺）的水平；蘇木素-伊紅（hematoxylin-eosin staining，HE）染色、Masson染色和普魯士藍(lán)染色觀察腎臟組織病理學(xué)變化；采用qRT-PCR檢測(cè)各組小鼠腎組織中GPX4和溶質(zhì)載體家族7成員11（solute carrier family 7 member 11，SLC7A11）的mRNA表達(dá)水平；采用Western blotting和免疫組化法檢測(cè)各組小鼠腎組織中GPX4和SLC7A11蛋白的表達(dá)水平。結(jié)果 與對(duì)照組比較，模型組小鼠體質(zhì)量、腎臟質(zhì)量和腎臟指數(shù)顯著升高（P＜0.01），血清Cre、BUN、UA和尿蛋白含量顯著升高（P＜0.01），腎組織ROS、MDA、LPO、總鐵和Fe²⁺的水平顯著升高（P＜0.01）、T-SOD活力均顯著降低（P＜0.01），腎臟GPX4和SLC7A11的mRNA和蛋白表達(dá)水平顯著降低（P＜0.01）；同時(shí)，模型組小鼠腎臟腎小管擴(kuò)張、腎小球纖維化并伴有鐵沉積。與模型組比較，莫諾苷給藥后顯著降低小鼠體質(zhì)量、腎臟質(zhì)量和腎臟指數(shù)（P＜0.05、0.01），顯著降低血清Cre、BUN、UA和尿蛋白的含量（P＜0.05、0.01），顯著降低腎組織ROS、MDA、LPO、總鐵和Fe²⁺的水平（P＜0.05、0.01），顯著升高腎組織T-SOD活力（P＜0.01）。同時(shí)，莫諾苷能夠減輕果糖飲食小鼠腎臟腎小管擴(kuò)張、腎小球纖維化和鐵沉積，顯著升高腎臟GPX4和SLC7A11的mRNA和蛋白表達(dá)水平（P＜0.05、0.01）。給予GPX4抑制劑干預(yù)后能夠明顯逆轉(zhuǎn)莫諾苷激活GPX4/SLC7A11信號(hào)通路抑制鐵死亡從而改善果糖飲食小鼠腎功能異常和腎臟損傷的作用。結(jié)論 莫諾苷能通過(guò)調(diào)控GPX4介導(dǎo)的鐵死亡從而改善果糖飲食小鼠腎損傷。

Objective To investigate the effect and mechanism of morroniside on ameliorating kidney injury in fructose-fed mice. Methods A total of 48 male C57BL/6 mice were randomly divided into control group, model group, allopurinol (25 mg/kg) group, morroniside low-dose (25 mg/kg) group, morroniside high-dose (50 mg/kg) group and morroniside (50 mg/kg) + glutathione peroxidase 4 (GPX4) inhibitor RSL3 (5 mg/kg) group. The mice in control group were given normal drinking water, and mice in the other groups were fed with 30% fructose water, while the mice in each treatment group were given corresponding drugs once a day for eight consecutive weeks. At the end of the experiment, serum, urine and kidney samples were collected from each group of mice, and body weight and kidney weight were recorded, the kidney index was calculated; The contents of creatinine (Cre), blood urea nitrogen (BUN) and uric acid (UA) in serum as well as the content of urinary protein in urine were detected, respectively. Meanwhile, the levels of reactive oxygen species (ROS), malondialdehyde (MDA), total-superoxide dismutase (T-SOD), lipid hydroperoxide (LPO), total iron, and ferrous ion (Fe²⁺) in kidney of mice in each group were detected. Histopathological changes in liver was observed by hematoxylin-eosin (HE) staining, Masson staining and Prussian Blue staining. qRT-PCR was used to detect the mRNA expressions of GPX4 and solute carrier family 7 member 11 (SLC7A11) in kidney tissue of mice in each group. Western blotting and immunohistochemistry were used to detect the protein expression levels of GPX4 and SLC7A11 in kidney tissue of mice in each group. Results Compared with control group, mice in model group showed significantly increased body weight, kidney weight and kidney index (P < 0.01), significantly increased contents of Cre, BUN, UA in serum and urinary protein (P < 0.01), significantly increased levels of ROS, MDA, LPO, total iron and Fe²⁺ in kidney tissues (P < 0.01), significantly decreased T-SOD activity (P < 0.01), and significantly reduced mRNA and protein expression levels of GPX4 and SLC7A11 in kidney tissues (P < 0.01). Meanwhile, the kidney of mice in model group showed kidney tubular dilatation, glomerular fibrosis and iron deposition. Compared with model group, morroniside significantly decreased body weight, kidney weight and kidney index (P < 0.05, 0.01), significantly decreased levels of Cre, BUN, UA in serum as well as the content of urinary protein in urine (P < 0.05, 0.01), significantly decreased the contents of ROS, MDA, LPO, total iron, and Fe²⁺ in kidney tissues (P < 0.05, 0.01), and significantly increased T-SOD activity in kidney tissues (P < 0.01). Meanwhile, morroniside was able to attenuate renal tubular dilatation, glomerular fibrosis and iron deposition, and significantly increased the mRNA and protein expression levels of GPX4 and SLC7A11 in kidney tissues (P < 0.05, 0.01). Intervention with GPX4 inhibitor (RSL3) could significantly reverse the effect of morroniside on inhibiting ferroptosis by activating GPX4/SLC7A11 signaling pathway to improve renal dysfunction and kidney injury in fructose-fed mice. Conclusion Morroniside can ameliorate kidney injury in fructose-diet mice by modulating GPX4-mediated ferroptosis.

R285.5

湖北省自然科學(xué)基金青年基金項(xiàng)目（2024AFB140）；中藥資源與中藥化學(xué)湖北省重點(diǎn)實(shí)驗(yàn)室開(kāi)放基金重點(diǎn)項(xiàng)目（KLRCCM2401）；武當(dāng)特色中藥研究湖北省重點(diǎn)實(shí)驗(yàn)室（湖北醫(yī)藥學(xué)院）開(kāi)放課題重點(diǎn)項(xiàng)目（WDCM2024001）