目的 建立一測多評（QAMS）結(jié)合化學(xué)模式識別與加權(quán)逼近理想排序（TOPSIS）法的不同產(chǎn)地小葉三點金Desmodiummicrophyllum總黃酮（DMTF）質(zhì)量綜合評價方法，初步探討其抗炎活性。方法 采用Thermo C₁₈色譜柱（250 mm× 4.6 mm,5 μm） ，流動相乙腈-0.1%甲酸梯度洗脫，體積流量0.8 mL·min^-1，柱溫20℃，檢測波長300、350 nm，進樣量10 μL。以異葒草苷為內(nèi)標，建立其與夏佛塔苷、葒草苷、異夏佛塔苷、木犀草素的相對校正因子，測定待測組分的含量，采用外標法（ESM）測定DMTF中這5種成分的含量，并比較兩種方法的差異，以驗證QAMS法的準確性和可靠性。結(jié)合化學(xué)模式識別和TOPSIS分析，評價不同產(chǎn)地DMTF質(zhì)量差異性。采用酶聯(lián)免疫吸附法（ELISA），觀察ig給予臨床等效劑量DMTF后采集的大鼠含藥血清對脂多糖（LPS，1 μg·mL^-1）誘導(dǎo)的巨噬細胞（RAW 264.7）釋放的白細胞介素（IL）-1β、IL-6含量的影響。結(jié)果 以異葒草苷為內(nèi)標，夏佛塔苷、葒草苷、異夏佛塔苷、木犀草素的相對校正因子分別為1.498、1.050、1.522、0.653，5種成分線性關(guān)系良好（r≥0.999 0），QAMS法與ESM所得結(jié)果無顯著性差異。化學(xué)模式識別法將10批樣品分為2類，葒草苷、異葒草苷、夏佛塔苷和異夏佛塔苷是影響DMTF質(zhì)量的主要差異性成分。加權(quán)TOPSIS法顯示10批DMTF歐氏貼近度為0.123 3～0.952 0，提示不同產(chǎn)地樣品存在一定差異，質(zhì)量優(yōu)劣排序和分類與化學(xué)模式識別聚類結(jié)果基本一致?？寡谆钚栽u價結(jié)果顯示，與對照組（10% FBS）相比，模型組IL-1β和IL-6水平顯著升高（P＜0.01） ；與模型組相比，10%含藥血清和陽性藥地塞米松（1 μg·mL^-1 LPS＋10 μg·mL^-1）均能顯著降低IL-1β和IL-6水平（P＜0.01）。結(jié)論 所建立的QAMS結(jié)合化學(xué)模式識別及加權(quán)TOPSIS法可用于評價不同產(chǎn)地DMTF質(zhì)量差異性； DMTF具有較好的體外抗炎活性，為小葉三點金質(zhì)量標準提升提供參考。

Objective To establish a comprehensive quality evaluation method for different origins of total flavonoids of Desmodium microphyllum (DMTF) by integrating QAMS, chemical pattern recognition and weighted TOPSIS, and to preliminarily explore its antiinflammatory activity. Methods Using a Thermo C₁₈ chromatographic column (250 mm × 4.6 mm, 5 μm), the mobile phase was acetonitrile-0.1% formic acid with a gradient elution, with a volume flow rate of 0.8 mL·min^-1, a column temperature of 20 ℃, and a detection wavelength of 300 and 350 nm. Using isoorientin as the internal standard and establishing its relative correction factor with schaftoside, orientin, isoschaftoside, and luteolin. The content of the analyte components was determined, and the content of these five components in DMTF was determined by the external standard method (ESM), and the differences between the two methods were compared to verify the accuracy and reliability of the QAMS method. Combined with chemical pattern recognition and TOPSIS analysis, the quality differences of DMTF from different origins were evaluated. The effect of drug-containing serum collected from rats after intragastric administration of the clinical equivalent dose of DMTF on the release of interleukin (IL)-1β and IL-6 by lipopolysaccharide (LPS, 1 μg·mL^-1)-induced macrophages (RAW264.7) was observed by enzyme-linked immunosorbent assay (ELISA). Results Using isoorientin as the internal standard, the relative correction factors of schaftoside, orientin, isoschaftoside, and luteolin were 1.498, 1.050, 1.522, 0.653, respectively. The linear relationship of the five components was good (r ≥ 0.999 0). There was no significant difference between the QAMS method and the ESM results. The chemical pattern recognition method classified 10 batches of samples into two categories, and orientin, isoorientin, schaftoside, and isoschaftoside were the main differential components affecting the quality of DMTF. The weighted TOPSIS method showed that the euclidean proximity degree of the 10 batches of DMTF was 0.123 3－ 0.952 0, suggesting that there were certain differences in the samples from different origins, and the ranking and classification of quality were basically consistent with the clustering results of chemical pattern recognition. The anti-inflammatory activity evaluation results showed that compared with the control group (10% FBS), the IL-1β and IL-6 levels in the model group were significantly increased (P < 0.01); compared with the model group, both the 10% drug-containing serum group and the positive drug group (1 μg·mL^-1 LPS + 10 μg·mL^-1 dexamethasone) could significantly reduce the levels of IL-1β and IL-6 (P < 0.01). Conclusion The QAMS combined with chemical pattern recognition and weighted TOPSIS method can be used to evaluate the quality differences of DMTF from different origins; DMTF has good in vitro anti-inflammatory activity, providing a reference for the improvement of the quality standard of D. microphyllum.

R284.1

中央引導(dǎo)地方科技發(fā)展資金資助項目（202201002）；廣西壯瑤藥重點實驗室開放課題（GXZYZZ202010）；廣西中醫(yī)藥大學(xué)第三批“岐黃工程”高層次人才團隊培育項目（202406）；廣西中醫(yī)藥管理局項目（GZZJ202029，GZZJ202030）；廣西中醫(yī)藥重點研究室項目（桂中醫(yī)藥科教發(fā)［2023］9號）；國家中醫(yī)藥管理局全國名老中醫(yī)藥專家傳承工作室建設(shè)項目-黃瑞松全國名老中醫(yī)藥專家傳承工作室（國中醫(yī)藥人教函［2022］75號）