黄芩甙对肝癌细胞BEL-7402形态学的影响

目的观察黄芩甙对肝癌细胞BEL-7402凋亡的影响,同时观察对肝癌细胞形态及超微结构、线粒体超微结构、线粒体膜电位和细胞内Ca^2＋的影响,探讨线粒体损伤在黄芩甙诱导肝癌细胞凋亡中的作用及可能的机制。方法应用细胞培养技术培养肝癌细胞BEL-7402,光镜、倒置显微镜、扫描电镜、透射电镜观察细胞形态及超微结构的变化尤其是线粒体的变化,应用流式细胞仪检测细胞凋亡百分率及线粒体膜电位、细胞内Ca^2＋的改变,免疫组化法检测细胞Bcl-2、Pax蛋白表达。结果黄芩甙诱导肝癌细胞BEL-7402凋亡呈剂量依赖关系,细胞形态、超微结构及线粒体超微结构出现明显改变,降低肝癌细胞线粒体膜电位,使细胞内Ca^2＋增加,细胞Pax表达增加,广泛分布于胞核和胞质中,Bcl-2表达减少。结论黄芩甙诱导肝癌细胞BEL-7402凋亡,线粒体损伤在黄芩甙诱导肝癌细胞凋亡中起重要作用,其机制可能为抑制肝癌细胞Bcl-2蛋白表达,促进Pax蛋白表达及细胞内Ca^2＋增加,激发线粒体膜通透性转运孔开放,线粒体跨膜电位降低,使肝癌细胞凋亡。     

HPLC with electrochemical detection to examine the pharmacokinetics of baicalin and baicalein in rat plasma after oral administration of a Kampo medicine

A highly sensitive method for determining baicalin and baicalein in rat plasma was developed using micro HPLC with electrochemical detection (muHPLC-ED). Peak heights for baicalin and baicalein were found to be linearly related to the amounts injected, ranging from 2.2 pg to 4.5 ng (r = 0.997) and from 1.4 pg to 2.7 ng (r = 0.997), respectively. The detection limits (signal/noise ratio = 3) of the current method for baicalin and baicalein were 0.89 and 0.54 pg, respectively. The concentrations of baicalin, baicalein, and their conjugates in rat plasma after oral administration of 2.0 mg/kg of Saiko-keishi-to (TJ-10) were determined. The glucuronide and sulfate forms of baicalin and baicalein in plasma were hydrolyzed enzymatically using beta-glucuronidase and sulfatase, respectively, and the hydrolyzed solutions were extracted with a 0.1-mol/L phosphoric acid-ethyl acetate mixture (1:1, v/v). Based on the time courses of the concentrations of the free and conjugated forms of baicalin and baicalein in rat plasma after oral administration of Saiko-keishi-to, the pharmacokinetic parameters of C(max), t(max), and AUC(0-6 h) were obtained.     

Zinc coupling potentiates anti-HIV-1 activity of baicalin

Baicalin (BA) has been shown with anti-HIV-1 activity. Zinc is a nutrient element. The anti-HIV-1 activity of zinc complex of baicalin (BA-Zn) in vitro was studied and compared with the anti-HIV-1 activities between BA and BA-Zn in the present study. Our results suggested that BA-Zn has lower cytotoxicity and higher anti-HIV-1 activity compared with those of BA in vitro. The CC50s of BA-Zn and BA were 221.52 and 101.73 microM, respectively. The cytotoxicity of BA-Zn was about 1.2-fold lower than that of BA. The BA and BA-Zn inhibited HIV-1 induced syncytium formation, HIV-1 p24 antigen and HIV-1 RT production. The EC50s of BA-Zn on inhibiting HIV-1 induced syncytium formation (29.08 microM) and RT production (31.17 microM) were lower than those of BA (43.27 and 47.34 microM, respectively). BA-Zn was more effective than BA in inhibiting the activities of recombinant RT and HIV-1 entry into host cells. Zinc coupling enhanced the anti-HIV-1 activity of baicalin.     

Flavone-catalyzed apoptosis in Scutellaria baicalensis

In response to mechanical damage, roots of Scutellaria baicalensis undergo cell death within 24 h. The flavone baicalein was identified as the factor regulating apoptosis in the damaged roots of S. baicalensis. Plant apoptosis is known to be triggered by oxidative damage of DNA through oxidative bursts, whereas baicalein causes apoptosis in Scutellaria cells by a copper-dependent oxidation of nuclear DNA without inducing an oxidative burst. S. baicalensis possesses an interesting system for quickly producing this apoptosis-inducing flavone in its cells. Intact Scutellaria cells contain little baicalein but store a large amount of baicalin (baicalein 7-O-β-D-glucuronide). Stress treatment of Scutellaria cells immediately initiates hydrolysis of baicalin by endogenous β-glucuronidase, and the resulting baicalein is immediately translocated to the nucleus, leading to apoptosis. Thus, S. baicalensis possesses a unique apoptosis-inducing system that is linked with metabolism of baicalin.     

黄芩苷作为一种抗生物制剂的过敏反应的研究

通过黄芩苷抗生物制剂过敏反应的研究,在不影响疫苗免疫效果的前提下,解决注射生物制剂引起的过敏反应问题,为生物制品与天然药物相结合开辟了一条新路。采用生物制品过敏试验常规检测方法。黄芩苷使用安全,抗过敏作用明显,具有抑制肥大细胞脱颗粒和降低毛细血管通透性作用。黄芩苷与生物制品相结合,初步试验结果表明有抗动物过敏反应的效果。     

Baicalin reverse AMPA receptor expression and neuron apoptosis in chronic unpredictable mild stress rats

Changes of a-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor expression could impacts the viability of neurons and brain levels of brain-derived neurotrophic factor (BDNF) expression in the key brain structures in the pathophysiology of depressive disorder. In the present study, chronic unpredictable mild stress (CUMS) degraded performance decreased AMPA receptor expression and increased neuron apoptosis. Treatment with baicalin (20, 40 mg/kg) significantly reversed these changes. This study demonstrates that baicalin has potent antidepressant effect, likely through up-regulated the expression of AMPA receptor and suppression neuron apoptosis in CUMS-treated rats.     

Baicalin,baicalein and wogonin inhibits high glucose-induced vascular inflammation in vitro and in vivo

Vascular inflammatory process has been suggested to play a key role in initiation and progression of atherosclerosis, a major complication of diabetes mellitus. Thus, in this study, we attempted to determine whether three structurally related polyphenols found in the Chinese herb Huang Qui, namely baicalin, baicalein, and wogonin, can suppress vascular inflammatory processes induced by high glucose (HG) in human umbilical vein endothelial cells (HUVECs) and mice. Data showed that HG induced markedly increased vascular permeability, monocyte adhesion, expressions of cell adhesion molecules (CAMs), formation of reactive oxygen species (ROS) and activation of nuclear factor (NF)-κB. Remarkably, all of the above mentioned vascular inflammatory effects of HG were attenuated by pretreatment with baicalin, baicalein, and wogonin. Vascular inflammatory responses induced by HG are critical events underlying development of various diabetic complications, therefore, our results suggest that baicalin, baicalein, and wogonin may have significant therapeutic benefits against diabetic complications and atherosclerosis. [BMB Reports 2015; 48(9): 519-524]     

丁香花蕾油作为微乳的油相促进黄芩苷透皮吸收的研究

目的:黄芩苷作为黄芩的主要成分之一,其制剂虽应用广泛,但存在较多缺点,如水溶液不稳定、口服吸收差、生物利用度不高及药物体内半衰期短等从而导致用药剂量大且频繁给药。本文目的是研究丁香花蕾精油及其微乳对黄芩苷体外经皮渗透性的影响。方法:采用Valia-Chien扩散池,以生理盐水为接受液,不同浓度的丁香花蕾精油以及以丁香花蕾精油作为微乳的油相及促进剂进行大鼠离体皮肤的体外渗透实验。结果:黄芩苷微乳的平均粒径为41.3±5.1 nm,多分散指数为0.18,表明该制剂粒径小且均一。微乳(不含丁香花蕾精油)、丁香花蕾精油及含丁香花蕾精油的微乳对黄芩苷均有不同程度的增渗作用。其中,含丁香花蕾精油的微乳对黄芩苷的促渗作用最强。结论:丁香花蕾精油以及以丁香花蕾精油作为微乳的油相对黄芩苷均有一定的促进渗透作用。     

黄芩苷对阪崎克罗诺杆菌生物膜的抑制作用

【目的】分析黄芩苷对阪崎克罗诺杆菌生物膜的抑制作用。【方法】采用XTT法评价黄芩苷对阪崎克罗诺杆菌起始粘附性及生物膜内细菌细胞活性的影响,并且采用荧光实时定量PCR(Quantitative real-time PCR,q RT-PCR)检测了阪崎克罗诺杆菌生物膜相关基因glp Q、nlp D、gsi B、deo B、lux S、sdi A的表达水平。【结果】黄芩苷对阪崎克罗诺杆菌的抑制效果呈剂量依赖型。黄芩苷对阪崎克罗诺杆菌的MIC80值为1 024 mg/L,该浓度的黄芩苷对阪崎克罗诺杆菌BAA-894和IQCC10423菌株生物膜的抑制率分别为83.7%和53.2%。浓度为2 048 mg/L的黄芩苷能够通过降低阪崎克罗诺杆菌的粘附性来抑制新生物膜的形成。另外,实时定量PCR结果表明黄芩苷可能通过下调阪崎克罗诺杆菌生物膜相关基因的表达来抑制其生物膜的形成。【结论】黄芩苷有可能被作为抗菌剂以预防和灭活阪崎克罗诺杆菌生物膜。