LC-MS/MS同时测定蓝芩口服液中6种化学成分的含量

目的 采用LC-MS/MS同时测定蓝芩口服液中6种化学成分（绿原酸、栀子苷、黄芩苷、盐酸小檗碱、黄芩素、汉黄芩素）的含量。方法 采用Fortis Xi C₁₈柱（50 mm×2.1 mm,1.7 μm）,以乙腈-0.1%甲酸水溶液为流动相,梯度洗脱,流速0.3 mL·min^-1。质谱采用电喷雾离子源,多反应监测方式进行检测。结果 6个成分在考察的浓度范围内具有良好的线性关系（r>0.999 1）;回收率（n=9）在94.7%~103.3%内,RSD均<3.3%;3个批次的样品测定结果均符合国家标准的要求,但批次间6个成分的含量差异较大。结论 本方法简便、快速、选择性好,灵敏度高,可用于蓝芩口服液的质量控制。     

Small interfering RNA target for long noncoding RNA PCGEM1 increases the sensitivity of LNCaP cells to baicalein

The objective of this study is to investigate the inhibitory effect and mechanism of long noncoding RNA PCGEM1 siRNA combined with baicalein on prostate cancer LNCaP cells. LNCaP cells transfected with small hairpin RNA lentiviral vector targeting PCGEM1 were constructed and their expression in LNCaP cells was absent. The stable cell line of LNCaP cells infected with LV3-shRNA-PCGEM1 was successfully constructed. In addition, LV3-shRNA-PCGEM1 was able to increase the baicalein-induced inhibitory effects on LNCaP cells, and the susceptibility was 2.3 fold higher than that of baicalein alone. LV3-shRNA-PCGEM1 combined with baicalein also inhibited the colony formation, increased G2 and S phase cells, inhibited the expression of PCGEM1, and induced autophagy of LNCaP cells. In summary, LV3-shRNA-PCGEM1 may improve the sensitivity of LNCaP cells to baicalein, and the molecular mechanism may be associated with the decrease of PCGEM1 expression and the induction of autophagy. Our findings provided an experimental basis for the combined treatment of Chinese traditional and Western medicine on prostate cancer in a clinical setting.     

miR-3,178 contributes to the therapeutic action of baicalein against hepatocellular carcinoma cells via modulating HDAC10

Hepatocellular carcinoma (HCC) is the most common type of hepatic malignancies with high mortality and poor prognosis. Baicalein, one of the major and bioactive flavonoids isolated from Scutellaria baicalensis Georgi, which is reported to have anti-proliferation effect in varying cancers, including HCC, whose underlying molecular mechanism is still largely unknown. In this study, we found that baicalein significantly inhibited proliferation and colony formation, blocked cell cycle, and promoted apoptosis in HCC cells MHCC-97H and SMMC-7721 in vitro and reduced tumor volume and weight in vivo. Increased microRNA (miR)-3,178 levels and decreased histone deacetylase 10 (HDAC10) expression were found in cells treated with baicalein and in patients' HCC tissues. HDAC10 was identified as a target gene of miR-3,178 by luciferase activity and western blot. Both baicalein treatment and overexpression of miR-3,178 could downregulate HDAC10 protein expression and inactivated AKT, MDM2/p53/Bcl2/Bax and FoxO3α/p27/CDK2/Cyclin E1 signal pathways. Not only that, knockdown of miR-3,178 could partly abolish the effects of baicalein and the restoration of HDAC10 could abated miR-3,178-mediated role in HCC cells. Collectively, baicalein inhibits cell viability, blocks cell cycle, and induces apoptosis in HCC cells by regulating the miR-3,178/HDAC10 pathway. This finding indicated that baicalein might be promising for treatment of HCC.     

Synergistic Antimicrobial Activity of Berberine Hydrochloride, Baicalein and Borneol against Candida albicans

Objective To determine the synergistic effects of berberine hydrochloride, baicalein,and borneol in different combinations on Candida albicans. Methods The broth microdilution method was used to determine the minimal inhibitory concentration(MIC) and minimal bactericidal concentration(MBC) of the three agents, and the checkerboard method was simultaneously used to determine the MIC and fractional inhibitory concentration index(FICI) of the combination of three antimicrobial agents to study their extracorporeal effects. Results Berberine hydrochloride was the most potent inhibitor of C. albicans(MIC and MBC of 0.160 and 0.640 mg/mL), followed by borneol(MIC and MBC of 0.320 and 0.640 mg/mL) and baicalein(MIC and MBC of 1.28 and 20.48 mg/mL). Moreover, the antifungal effect of the combination was significantly stronger than that tested alone. Further in vivo study showed that the mortality rate of tainted mice reduced over 50% compared with the control group. Conclusion The results of experiments in vitro and in vivo indicate the synergistic effect of the combination of three antimicrobial agents on C. albicans, which can make reference for the future clinical treatment.     

Baicalein,a component of banxia xiexin decoction,alleviates CPT-11-induced gastrointestinal dysfunction by inhibiting ALOX15-mediated ferroptosis

Baicalein, one of the active ingredients of banxia xiexin decoction, has good therapeutic efficacy in treating diarrhea and improving gastrointestinal dysfunction. The role and mechanism of Baicalein on irinotecan (CPT-11)-induced gastrointestinal dysfunction are the focus of this study. Concretely, CPT-11 induced delayed diarrhea rat model and intestinal epithelial cell (IEC)-6 cell injury model with Baicalein treatment as needed. Colonic pathological changes were analyzed by hematoxylin–eosin staining, and inflammatory factor expressions in serum were determined by enzyme-linked immunosorbent assay. Immunohistochemistry and western blot were performed to quantify ferroptosis-related protein expressions. Thiobarbituric acid reactive substances (TBARS) kits and colorimetric assay kit were applied to detect lipid peroxidation levels and Fe²⁺ content, respectively. In vitro experiments also included quantitative real-time polymerase chain reaction, cell counting kit-8, and C11 BODIPY staining. CPT-11 induced aggravation of intestinal tissue damage, inflammatory factor release, Fe²⁺ accumulation, upregulation of lipid peroxidation and 15-Lipoxygenase (ALOX15) expression, and downregulation of glutathione peroxidase 4 (Gpx4) and SLC7A11 in vivo in rats; however, Baicalein dose-dependently reversed the effects of CPT-11. Baicalein elevated cell viability, reduced lipid peroxidation and Fe²⁺ accumulation, and elevated Gpx4 and SLC7A11 levels, whereas ALOX15 overexpression reversed the effects of Baicalein on a CPT-11-induced IEC-6 cell injury model. In conclusion, Baicalein plays a mitigating role in CPT-11-induced delayed diarrhea via ALOX15-mediated ferroptosis.     

多指标综合评价结合层次分析法优化黄芩的微波酒蜜炮制工艺

目的 优选黄芩的微波酒蜜制工艺。方法 采用单因素试验优选酒蜜比、闷润时间、微波功率、微波时间。以微波工艺中的酒蜜比、闷润时间、微波功率、微波时间为因素，以黄芩苷、汉黄芩苷、黄芩素、汉黄芩素质量分数为指标，采用层次分析法确定各指标权重系数，综合加权评分为评价指标，采用正交试验优选黄芩的微波酒蜜制工艺。结果 最佳微波酒蜜制工艺为取生黄芩饮片适量，加入25%的酒蜜混合辅料（酒蜜比为10：15），置密闭容器内闷润60 min，在300 W功率下微波3 min。结论 方法重复性和稳定性良好，可用于微波酒蜜炮制黄芩。     

基于网络药理学和分子对接研究黄芩治疗白癜风的作用机制

目的 利用网络药理学方法研究黄芩治疗白癜风的作用靶点和机制。方法 使用中药系统药理学数据库与分析平台(TCMSP)筛选得到黄芩中的活性成分及其作用靶点,通过SwissTargetPrediction数据库检索补充各个活性成分的作用靶点;通过OMIM、GeneCards数据库获取白癜风的相关靶点,与活性成分作用靶点取交集;将黄芩活性成分和白癜风的交集靶点导入String数据库构建靶点间的蛋白相互作用(PPI)网络;利用Cytoscape 3.9.1软件,构建"黄芩-活性成分-白癜风靶点"网络并筛选核心靶点;使用Metascape数据库对潜在作用靶点进行基因本体论(GO)功能富集分析和京都基因与基因组百科全书(KEGG)通路富集分析;使用AutoDockTools 1.5.7软件进行分子对接验证并使用PyMol软件对结果进行可视化处理。结果 共筛选得到黄芩素、汉黄芩素、刺槐素、木蝴蝶素a、表小檗碱、去甲汉黄芩素、5,7,4''-三羟基-8-甲氧基黄酮等36个活性成分,作用于3 868个潜在靶点,白癜风相关靶点1 349个,黄芩-白癜风的交集靶点113个;PPI网络分析得到肿瘤蛋白P53(TP53)、丝氨酸/苏氨酸蛋白激酶1(AKT1)、信号转导和转录激活因子3(STAT3)、肿瘤坏死因子(TNF)、丝裂原活化蛋白激酶1(MAPK1)、白细胞介素-6(IL-6)、胱天蛋白酶3(CASP3)、雌激素受体(ESR1)共8个核心作用靶点;GO和KEGG富集分析显示,黄芩治疗白癜风主要涉及胰腺癌、内分泌抵抗、神经营养素信号通路、细胞凋亡、乙型肝炎、细胞衰老等信号通路。分子对接结果显示黄芩素、汉黄芩素、刺槐素、木蝴蝶素a、表小檗碱、去甲汉黄芩素、5,7,4''-三羟基-8-甲氧基黄酮7个核心成分与核心靶点具有较好的结合能。结论 黄芩可能通过调控内分泌、神经、细胞衰老和凋亡以及炎症等方面发挥治疗白癜风的作用,为后续研究黄芩治疗白癜风提供参考。     

基于网络药理学和指纹图谱的安宫牛黄丸质量标志物预测与分析

目的 通过网络药理学与指纹图谱预测安宫牛黄丸中的潜在质量标志物（Q-Marker）。方法 应用网络药理学筛选和分析安宫牛黄丸活性成分及临床适应证（卒中、高热昏迷、脑炎、脑出血及癫痫）的作用靶点和通路，并寻找关键活性成分；应用超高效液相色谱（UPLC）构建安宫牛黄丸的指纹图谱，结合网络药理学预测其潜在的Q-Marker，并使用AutoDock Vina软件对潜在的Q-Marker与关键靶点进行分子对接验证。结果 收集得到安宫牛黄丸活性成分128个，经过筛选得到10个核心靶点（STAT3、AKT1、MAPK1等）和7个关键活性成分（小檗碱、槲皮素、汉黄芩素、黄芩素、熊果酸、黄芩苷及麝香酮），涉及炎症反应、细胞免疫、脂质代谢等相关机制。安宫牛黄丸的UPLC指纹图谱，标定22个共有峰，并指认出15个色谱峰，结合网络药理学筛选出的关键活性成分初步预测4个成分为其潜在的Q-Marker，分别是小檗碱、黄芩苷、黄芩素、汉黄芩素，涉及的关键生物通路包括AGEs-RAGE通路、PI3K-Akt通路以及MAPK通路等。结论 借助网络药理学结合UPLC指纹图谱分析预测得到安宫牛黄丸潜在的Q-Marker分别为小檗碱、黄芩苷、黄芩素、汉黄芩素，为其全面质量控制提供依据。     

Fabrication of baicalein-encapsulated zeolitic imidazole framework as a novel nanocomposited wound closure material to persuade pH-responsive healing efficacy in post-caesarean section wound care

Baicalein is a plant-derived compound, it has widespread attention among biomedical researchers due to its biocompatibility and efficient biological activities. But it has non-solubility in physiological conditions and short bioavailability in the clinical process. The Discovery of a stimuli-based drug delivery system (DDS) is a perfect strategy to improve the bioactivity of baicalin in post-caesarean section wound care. We prepared a baicalein-encapsulated pH-responsive DDS with a zeolite imidazole framework-8 (ZIF-8) as a carrier. We investigated the material characteristics of DDS in terms of crystal structure, surface area morphology, particle size, FTIR, UV–vis, Powder XRD, and BET analysis techniques. The in vitro cytotoxicity, biocompatibility, and cell proliferation properties of BA@ZIF-8 were assessed against human fibroblast L929 cells. In vitro studies showed that BA@ZIF-8 nanocomposite significantly enhanced the biocompatibility against L929 cells after 72 h post-exposure. The pH-responsive drug release kinetics shows excellent baicalein release under acidic conditions and without unwanted drug release in physiological conditions. Wound scratch assay results revealed, that BA@ZIF-8 nanocomposite-treated cells exhibit the fastest cell proliferation and migration process in a very short period. In the antibacterial activity study, nanocomposite exposed dose depended on inhibition against wound infectious pathogens. The overall study signifies that BA@ZIF-8 nano-DDSs are effective and suitable DDS for stimuli-based post-caesarean section wound care.