目的 應(yīng)用正交試驗(yàn)設(shè)計(jì)，結(jié)合層次分析法（analytic hierarchy process，AHP）-熵權(quán)法，優(yōu)化經(jīng)典名方黃連解毒湯（Huanglian Jiedu Decoction，HJD）的提取工藝，并通過(guò)細(xì)胞實(shí)驗(yàn)驗(yàn)證其抗炎效果，為HJD的合理化提取提供參考依據(jù)。方法 以HJD中代表成分小檗堿、黃芩苷、梔子苷和綠原酸的含量以及上清液和自沉淀干膏率為考察指標(biāo)；以加水量、提取時(shí)間和提取次數(shù)為關(guān)鍵工藝參數(shù)；采用正交試驗(yàn)結(jié)合AHP-熵權(quán)法確定各指標(biāo)的權(quán)重系數(shù)，計(jì)算綜合評(píng)分。通過(guò)脂多糖（lipopolysaccharide，LPS）誘導(dǎo)的小鼠小膠質(zhì)BV2細(xì)胞炎癥模型，探討HJD提取工藝綜合評(píng)分與其抗炎活性的相關(guān)性，并驗(yàn)證優(yōu)選的提取工藝的抗炎穩(wěn)定性。結(jié)果HJD優(yōu)化提取工藝為10倍水量、提取30 min、提取3次。取綜合評(píng)分為7.67、9.82和12.05的提取工藝制備HJD，分別命名為H1、H2、H3，進(jìn)行藥效學(xué)驗(yàn)證。結(jié)果顯示，與對(duì)照組相比，LPS誘導(dǎo)BV2細(xì)胞分泌NO、腫瘤壞死因子-α（tumor necrosis factor-α，TNF-α）和白細(xì)胞介素-6（interleukin-6，IL-6）；與LPS組相比，H1、H2、H3均能降低各組的炎癥因子水平，且相關(guān)性分析表明，評(píng)分較高的提取工藝制備的HJD可能具有更好的抗炎潛力。進(jìn)一步對(duì)H3進(jìn)行抗炎活性驗(yàn)證，結(jié)果顯示H3在100、200、400 μg/mL劑量下均能降低LPS誘導(dǎo)的炎癥因子水平，并呈現(xiàn)良好的劑量效應(yīng)關(guān)系。結(jié)論 正交試驗(yàn)結(jié)合AHP-熵權(quán)法篩選的HJD提取工藝準(zhǔn)確可靠，且穩(wěn)定可行，研究結(jié)果可為HJD后續(xù)開(kāi)發(fā)和應(yīng)用提供科學(xué)依據(jù)。

Objective To optimize the extraction process of the classical herbal formula Huanglian Jiedu Decoction (HJD, 黃連解毒湯) by orthogonal experimental design combined with analytic hierarchy process (AHP) and entropy weight method (EWM), and validate its anti-inflammatory effects through cellular assays, so as to provide a reference for the rational extraction of HJD. Methods The contents of representative components in HJD, including berberine, baicalin, geniposide, and chlorogenic acid, as well as the mass of the supernatant and precipitated dry extract rate, were used as indicators. Key extraction parameters, such as the amount of water added, extraction time, and number of extractions, were selected. An orthogonal experiment combined with AHP-EWM was used to determine the comprehensive weight coefficients for each indicator, and a total score was calculated. The correlation between the comprehensive score of the extraction process of HJD and its anti-inflammatory activity was explored using the LPS-induced BV2 microglial cell inflammation model, and the anti-inflammatory stability of the optimized extraction process was verified. Results The optimized extraction process for HJD was determined to be 10 times the amount of water, with an extraction time of 30 min and three extractions. Extraction processes with comprehensive scores of 7.67, 9.82, and 12.05 were used to prepare HJD, named H1, H2, and H3, respectively, for pharmacological validation. The results showed that, compared to the blank control group, LPS-induced BV2 cells secreted NO, tumor necrosis factor-α (TNF-α), and interleukin-6 (IL-6). Compared to the LPS group, H1, H2, and H3 all reduced the levels of inflammatory cytokines. Furthermore, correlation analysis revealed that HJD with higher extraction process scores exhibited stronger anti-inflammatory effects. Anti-inflammatory activity was further validated for H3, and the results demonstrated that H3, at three doses (100, 200, 400 μg/mL), significantly reduced LPS-induced inflammatory cytokine levels, demonstrating a good dose-response relationship. Conclusion The extraction process of HJD optimized by the orthogonal experiment combined with AHP-EWM is accurate, reliable, stable, and feasible. The research results can provide reference for subsequent studies on development and application of HJD.

R283.6

國(guó)家自然科學(xué)基金資助項(xiàng)目（82174358）；四川省中醫(yī)藥管理局資助項(xiàng)目（2023MS481）