[关键词]
[摘要]
目的 优选盐麸泽泻Alismatis Rhizoma的炮制工艺,并探究其对脂多糖诱导的小鼠小肠炎症的改善作用及其机制。方法 以外观性状、醇溶性浸出物含量、23-乙酰泽泻醇B、23-乙酰泽泻醇C、24-乙酰泽泻醇A和泽泻醇B作为评价指标,对炮制过程中盐水用量、麦麸用量、炒制时间和炒制温度进行考察,通过层次分析(analytic hierarchy process,AHP)-CRITIC(criteria importance though intercrieria correlation)复合加权法确定各评价指标的权重系数,联合单因素实验与Box-Behnken设计-响应面法(Box-Behnken design-response surface method,BBD-RSM)确定盐麸泽泻炮制工艺的最佳参数。基于优选工艺制备样品后,通过ip脂多糖建立小鼠小肠炎症模型,系统评价盐麸泽泻对小鼠的行为学特征与体质量动态变化、血液指标的影响、小肠组织病理学改变、回肠组织中肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-1β(interleukin-1β,IL-1β)、IL-6的释放水平以及组织中磷脂酰肌醇3激酶(phosphatidylinositol 3-kinase,PI3K)、蛋白激酶B(protein kinase B,Akt)、磷酸化蛋白激酶B(phosphorylated protein kinase B,p-Akt)、磷酸化核因子κB抑制蛋白(phosphorylated inhibitor of nuclear factor kappa-B,p-IκB)和核因子κB p65亚基(nuclear factor kappa-B p65 subunit,NF-κB p65)蛋白质的表达水平。结果 盐麸泽泻最佳炮制工艺参数:每100克泽泻饮片用15%盐水溶液闷润,晾干后与50 g麦麸在160℃炒制3 min。药效实验结果表明,盐麸泽泻水煎液可显著减轻脂多糖ip引起的小鼠体质量下降,血液指标的异常,回肠组织的损伤、炎症因子(TNF-α、IL-6、IL-1β)浓度升高以及组织中PI3K、p-Akt/Akt、p-Akt、p-IκB和NF-κB p65蛋白的表达水平上升。结论 优选出的盐麸泽泻工艺操作简便,稳定可行,按照此工艺制备的盐麸泽泻,可有效改善脂多糖诱导的小鼠小肠炎症,其作用机制可能与PI3K-Akt/NF-κB通路有关。
[Key word]
[Abstract]
Objective To optimize the processing technology of Zexie (Alismatis Rhizoma, AR) with salt bran, and to explore its improvement effect and mechanism on lipopolysaccharide (LPS)-induced intestinal inflammation in mice. Methods The appearance traits, alcohol-soluble extract content, 23-acetyl alisol B, 23-acetyl alisol C, 24-acetyl alisol A and alisol B were used as evaluation indexes to investigate the amount of salt water, the amount of wheat bran, the frying time and the frying temperature during the processing. The weight coefficient of each evaluation index was determined by analytic hierarchy process (AHP)-criteria importance though intercrieria correlation (CRITIC) comprehensive weighting method, and the optimal parameters of the processing technology of salt bran Alismatis Rhizoma were determined by single factor experiment and response surface method. After the samples were prepared based on the optimal process, the intestinal inflammation model of mice was established by intraperitoneal injection of LPS. The effects of salt bran Alismatis Rhizoma on behavioral characteristics, dynamic changes of body weight, blood indexes, histopathological changes of small intestine, release levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and IL-6 in ileum tissues, and protein expression levels of phosphatidylinositol 3-kinase (PI3K), protein kinase B (Akt), phosphorylated protein kinase B (p-Akt), phosphorylated inhibitor of nuclear factor kappa-B (p-IκB) and nuclear factor kappa-B p65 subunit (NF-κB p65) in tissues were systematically evaluated. Results The optimum processing parameters of Alismatis Rhizoma with salt bran were as follows: per 100 g of Alismatis Rhizoma decoction pieces were moistened with 15% salt water solution dried and stir-fried with 50 g wheat bran at 160 ℃ for 3 min. The results of pharmacodynamic experiments showed that the water decoction of Alismatis Rhizoma with salt bran could significantly reduce the weight loss of mice caused by intraperitoneal injection of LPS, the abnormality of blood indexes, the damage of ileum tissue, the increase of inflammatory factors (TNF-α, IL-6, IL-1β) and the expression of PI3K, p-Akt/Akt, p-Akt, p-IκB and NF-κB p65 protein in tissues. Conclusion The optimized salt bran Alismatis Rhizoma process is simple, stable and feasible. The salt bran Alismatis Rhizoma prepared according to this process can effectively improve LPS-induced intestinal inflammation in mice, and its mechanism may be related to the PI3K-Akt/NF-κB pathway.
[中图分类号]
R283.6
[基金项目]
国家重点研发计划“中医药现代化研究”重点专项(2023YFC3504205);国家自然科学基金资助项目(82360771);特色炮制技术规律发掘——麸制(GZY-KJS-2022-051);江西省“双千人才”计划项目[赣才办字(2019)9号];江西省中医药产业科技创新联合体产业链协同攻关项目(20224BBG72001);江西省自然科学基金项目(20224ACB206048);中药炮制技术传承创新团队(CXTD22003);中药炮制传承创新与转化江西省重点实验室(2024SSY07091);“樟帮”特色中药饮片炮制规范标准研究(20223AAG02021)