目的 对黄芪六一汤中黄芪总皂苷、黄芪总黄酮、黄芪多糖及甘草酸、甘草总黄酮、甘草多糖分别进行提取、分离，并筛选具有防治糖尿病肾病作用的药效组分。方法 单因素法筛选树脂型号、上样浓度和体积、洗脱溶剂及用量建立大孔吸附树脂法对黄芪总皂苷、总黄酮进行富集分离；运用碱液萃取法对甘草酸、甘草总黄酮进行分离，采用水提醇沉法对黄芪多糖、甘草多糖进行纯化。12周龄雄性db/m小鼠8只作为对照组，将12周龄雄性db/db小鼠随机分为模型组、黄芪六一汤和各组分的高、低剂量（临床等效剂量的4、1倍）组，小鼠一般情况及血生化指标以盐酸罗格列酮片（RSG）为阳性对照；尿白蛋白（U-Alb）水平以缬沙坦分散片（API）为阳性对照，每组8只。各组小鼠分别在ig给药12周前、后，称质量。禁食12 h，自由饮水，收集24 h尿液，马斯亮蓝法检测24 h U-Alb浓度；尾静脉采血测空腹血糖（FBG）；摘取眼球取血，采用全自动生化分析仪检测血清血肌酐（Scr）、尿素氮（BUN）、三酰甘油（TG）、总胆固醇（TC）含量。对照组、模型组及黄芪六一汤高、低剂量组小鼠肾组织经固定、石蜡包埋切片，Masson染色后，光学显微镜下观察。结果 制得的黄芪总黄酮、黄芪总皂苷、黄芪多糖、甘草酸、甘草总黄酮、甘草多糖质量分数分别为（70.58±2.16）%、（72.97±1.06）%、（67.12±2.60）%、（81.02±1.04）%、（53.56±1.63）%、（64.62±1.27）%。与模型组比较，给予黄芪六一汤治疗后的各组小鼠肾纤维化程度较轻；与给药前比较，模型组小鼠体质量显著减轻（P＜0.05），甘草总黄酮高、低剂量和甘草多糖低剂量组体质量显著降低（P＜0.05、0.01），其他给药组无显著差异；与模型组比较，黄芪六一汤、黄芪总皂苷、黄芪多糖、黄芪总黄酮、甘草酸FBG、24 h U-Alb显著降低（P＜0.05、0.01），黄芪总皂苷、黄芪六一汤、黄芪总黄酮、甘草酸组Scr显著降低（P＜0.05、0.01），黄芪六一汤、黄芪多糖、甘草酸、黄芪总黄酮组BUN显著降低（P＜0.05、0.01），黄芪六一汤、黄芪总皂苷、甘草酸、黄芪多糖高剂量组TG显著降低（P＜0.05、0.01），黄芪六一汤、黄芪总皂苷、黄芪总黄酮、黄芪多糖组TC显著降低（P＜0.05、0.01）。结论 各组分确定的制备工艺稳定、科学、可行；黄芪总皂苷、黄芪总黄酮、黄芪多糖、甘草酸可能是黄芪六一汤防治糖尿病肾病的药效物质基础。

Objective The total astragaloside (TA), total flavonoids of astragalus (TFA), astragalus polysaccharide (AP), glycyrrhizic acid (GA), total flavonoids of glycyrrhiza (TFG) and glycyrrhiza polysaccharide (GP) in Huangqi Liuyi Decoction (HLD) were extracted and separated respectively, and the effective components with the effect of preventing and treating diabetes nephropathy were screened. Methods Single factor method was used to screen resin type, sample concentration and volume, elution solvent and dosage. A macroporous adsorption resin method was established to enrich and separate TA and TFA. Using alkaline extraction method to separate GA and TFG, purifying AP and GP using water extraction and alcohol precipitation. Eight 12 weeks old male db/m mice were used as control group. The 12 weeks old male db/db mice were randomly divided into model group, and high-dose and low-dose groups of HLD and each component (4 or 1 times the clinical equivalent dose), the general situation and blood biochemical indicators of mice were compared with rosiglitazone hydrochloride tablets (RSG) as positive controls, urinary albumin (U-Alb) levels were measured using valsartan dispersible tablets (API) as a positive control, with eight mice in each group. Each group of mice was weighed before and after 12 weeks of ig administration. Fasting for 12 hours, drinking water freely, collecting urine for 24 hours, and detecting the concentration of U-Alb for 24 hours using the Maas Brilliant Blue method. Tail vein blood collection for FBG measurement. Eye extraction and blood collection were performed, and serum creatinine (Scr), urea nitrogen (BUN), triglycerides (TG), and total cholesterol (TC) levels were detected using a fully automated biochemical analyzer. The renal tissues of mice in the control group, model group, and high-dose and low-dose groups of HLD were fixed, paraffin embedded, and sectioned. After Masson staining, they were observed under an optical microscope. Results The mass fraction of TA, TFA, AP, GA, TFG, and GP were (70.58 ± 2.16)%, (72.97 ± 1.06)%, (67.12 ± 2.60)%, (81.02 ± 1.04)%, (53.56 ± 1.63)%, and (64.62 ± 1.27)%. Compared with model group, the degree of renal fibrosis in each group of mice treated with HLD was milder. Compared with before administration, the body weight of the model group mice decreased significantly (P < 0.05), while the body weight of the high and low doses of TFG and low doses of GP decreased significantly (P < 0.05, 0.01), and there was no significant difference in the other administration groups. Compared with the model group, FBG and 24-hour U-Alb in HLD, TA, AP, TFA, GA group were significantly reduced (P < 0.05, 0.01), while the Scr of TA, HLD, TFA, and GA groups was significantly reduced (P < 0.05, 0.01), the BUN of HLD, AP, GA, and TFA groups was significantly reduced (P < 0.05, 0.01), the TG level of HLD, TA, GA, and AP groups also showed a significant decrease (P < 0.05, 0.01). TG significantly decreased (P < 0.05, 0.01), while TC significantly decreased (P < 0.05, 0.01) in the HLD, TA, TFA, and AP groups. Conclusion The TA, TFA, GA and AP are the main ingredients in HLD against diabetic nephropathy.

R285.5

贵州省普通高等学校青年科技人才成长项目（黔教合KY字［2022］257号）；贵州省科技计划项目（黔科合基础-ZK［2023］一般416）；2023年度贵州中医药大学学术新苗项目（贵科合学术新苗［2023］-13号）；贵州省卫生健康委科学技术基金项目（黔卫健函［2024］24号）