目的 對(duì)黃芪六一湯中黃芪總皂苷、黃芪總黃酮、黃芪多糖及甘草酸、甘草總黃酮、甘草多糖分別進(jìn)行提取、分離，并篩選具有防治糖尿病腎病作用的藥效組分。方法 單因素法篩選樹(shù)脂型號(hào)、上樣濃度和體積、洗脫溶劑及用量建立大孔吸附樹(shù)脂法對(duì)黃芪總皂苷、總黃酮進(jìn)行富集分離；運(yùn)用堿液萃取法對(duì)甘草酸、甘草總黃酮進(jìn)行分離，采用水提醇沉法對(duì)黃芪多糖、甘草多糖進(jìn)行純化。12周齡雄性db/m小鼠8只作為對(duì)照組，將12周齡雄性db/db小鼠隨機(jī)分為模型組、黃芪六一湯和各組分的高、低劑量（臨床等效劑量的4、1倍）組，小鼠一般情況及血生化指標(biāo)以鹽酸羅格列酮片（RSG）為陽(yáng)性對(duì)照；尿白蛋白（U-Alb）水平以纈沙坦分散片（API）為陽(yáng)性對(duì)照，每組8只。各組小鼠分別在ig給藥12周前、后，稱(chēng)質(zhì)量。禁食12 h，自由飲水，收集24 h尿液，馬斯亮藍(lán)法檢測(cè)24 h U-Alb濃度；尾靜脈采血測(cè)空腹血糖（FBG）；摘取眼球取血，采用全自動(dòng)生化分析儀檢測(cè)血清血肌酐（Scr）、尿素氮（BUN）、三酰甘油（TG）、總膽固醇（TC）含量。對(duì)照組、模型組及黃芪六一湯高、低劑量組小鼠腎組織經(jīng)固定、石蠟包埋切片，Masson染色后，光學(xué)顯微鏡下觀(guān)察。結(jié)果 制得的黃芪總黃酮、黃芪總皂苷、黃芪多糖、甘草酸、甘草總黃酮、甘草多糖質(zhì)量分?jǐn)?shù)分別為（70.58±2.16）%、（72.97±1.06）%、（67.12±2.60）%、（81.02±1.04）%、（53.56±1.63）%、（64.62±1.27）%。與模型組比較，給予黃芪六一湯治療后的各組小鼠腎纖維化程度較輕；與給藥前比較，模型組小鼠體質(zhì)量顯著減輕（P＜0.05），甘草總黃酮高、低劑量和甘草多糖低劑量組體質(zhì)量顯著降低（P＜0.05、0.01），其他給藥組無(wú)顯著差異；與模型組比較，黃芪六一湯、黃芪總皂苷、黃芪多糖、黃芪總黃酮、甘草酸FBG、24 h U-Alb顯著降低（P＜0.05、0.01），黃芪總皂苷、黃芪六一湯、黃芪總黃酮、甘草酸組Scr顯著降低（P＜0.05、0.01），黃芪六一湯、黃芪多糖、甘草酸、黃芪總黃酮組BUN顯著降低（P＜0.05、0.01），黃芪六一湯、黃芪總皂苷、甘草酸、黃芪多糖高劑量組TG顯著降低（P＜0.05、0.01），黃芪六一湯、黃芪總皂苷、黃芪總黃酮、黃芪多糖組TC顯著降低（P＜0.05、0.01）。結(jié)論 各組分確定的制備工藝穩(wěn)定、科學(xué)、可行；黃芪總皂苷、黃芪總黃酮、黃芪多糖、甘草酸可能是黃芪六一湯防治糖尿病腎病的藥效物質(zhì)基礎(chǔ)。

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

貴州省普通高等學(xué)校青年科技人才成長(zhǎng)項(xiàng)目（黔教合KY字［2022］257號(hào)）；貴州省科技計(jì)劃項(xiàng)目（黔科合基礎(chǔ)-ZK［2023］一般416）；2023年度貴州中醫(yī)藥大學(xué)學(xué)術(shù)新苗項(xiàng)目（貴科合學(xué)術(shù)新苗［2023］-13號(hào)）；貴州省衛(wèi)生健康委科學(xué)技術(shù)基金項(xiàng)目（黔衛(wèi)健函［2024］24號(hào)）