目的 通过肾脏代谢组学技术，揭示知母皂苷治疗糖尿病肾病的潜在作用机制。方法 SD雄性大鼠40只，随机分为对照组、模型组、二甲双胍(阳性药，65 mg·kg^-1)组、知母皂苷(30 mg·kg^-1)组，每组8只。除对照组外，各组大鼠喂养高脂饲料，连续28 d,ip链脲佐菌素（STZ）40 mg·kg^-1用于制备2型糖尿病（T2DM）大鼠模型。于造模成功后，给药组ig给予知母皂苷或二甲双胍，对照组和模型组大鼠每天ig蒸馏水，连续给药8周。给药结束后，称大鼠体质量，使用血糖仪进行尾静脉血糖检测；生化仪测定血清三酰甘油（TG）、总胆固醇（TC）、血肌酐（Scr）、尿素氮（BUN）水平；苏木精-伊红（HE）染色观察肾组织病理变化；采用超高效液相色谱串联飞行时间质谱分析技术（UPLC-Q-TOF-MS）对对照组、模型组、知母皂苷组大鼠肾脏代谢组学进行分析，采用主成分分析法探寻潜在的关键性生物标志物，联合人类代谢组学数据库（HMDB）和京都基因与基因组百科全书数据库（KEGG）找出对应的相关性代谢通路。结果 与模型组比较，二甲双胍、知母皂苷组大鼠体质量、血糖水平显著降低（P<0.05、0.01），血清Scr、BUN、TC、TG水平均显著降低（P<0.05、0.01），肾小管扩张、间质纤维化、血管淤血等情况均有改善。糖尿病肾病模型大鼠肾脏样本中发现21个潜在生物标志物，涉及18条相关代谢通路，其中甘油磷脂代谢、苯丙氨酸/酪氨酸/色氨酸生物合成、亚油酸代谢为主要的代谢途径。结论 知母皂苷可能通过调节甘油磷脂代谢、苯丙氨酸/酪氨酸/色氨酸生物合成、亚油酸代谢治疗糖尿病肾病。

Objective To reveal the potential mechanism of action of Anemarrhena saponin in the treatment of diabetic nephropathy through renal metabolomics technology. Methods Forty male SD rats were randomly divided into the control group, model group, metformin(positive drug, 65 mg·kg^-1) group, and Anemarrhena saponin(30 mg·kg^-1) group, with 8 rats in each group. Except for the control group, all groups were fed a high-fat diet for 28 consecutive days and ip injected with streptozotocin（STZ） at 40 mg·kg^-1 to establish a type 2 diabetic（T2DM） rat model. After successful modeling, the drug groups were ig administered Anemarrhena saponin or metformin, while the control group was ig administered distilled water daily for 8 consecutive weeks. After the administration period, the body weight of the rats was measured, and blood glucose levels were detected by a blood glucose meter through the tail vein. Serum triglycerides（TG）, total cholesterol（TC）, serum creatinine（Scr）, and blood urea nitrogen（BUN） levels were determined by a biochemical analyzer. Renal tissue pathological changes were observed by hematoxylin-eosin（HE） staining. Ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry（UPLC-Q-TOF-MS） was used to analyze the renal metabolomics of the control group, model group, and Anemarrhena saponin group. Principal component analysis was used to explore potential key biomarkers, and the Human Metabolome Database（HMDB） and Kyoto Encyclopedia of Genes and Genomes（KEGG） were used to identify the corresponding metabolic pathways. Results Compared with the model group, the body weight and blood glucose levels of the metformin and Anemarrhena saponin groups were significantly reduced（P ＜ 0.05, 0.01）, and the levels of serum Scr, BUN, TC, and TG were significantly decreased（P ＜ 0.05, 0.01）. The conditions of renal tubular dilation, interstitial fibrosis, and vascular congestion were improved. Twenty-one potential biomarkers were identified in the renal samples of diabetic nephropathy model rats, involving 18 related metabolic pathways, among which glycerophospholipid metabolism, phenylalanine/tyrosine/tryptophan biosynthesis, and linoleic acid metabolism were the main metabolic pathways. Conclusion Anemarrhena saponin may treat diabetic nephropathy by regulating glycerophospholipid metabolism, phenylalanine/tyrosine/tryptophan biosynthesis, and linoleic acid metabolism.

R285.5

2024年度黑龙江省卫生健康委科技计划资助项目（20241313050500）