汉黄芩素对肺癌肿瘤细胞生长和转移抑制作用机制的研究

目的:探讨明胶酶(MMP-2)、表皮生长因子受体酪氨酸蛋白激酶(EGFR-TPK)和氨肽酶N(APN)抑制剂汉黄芩素对肺癌细胞的生长和转移抑制机制。方法:本文采用酶抑制学方法,研究了汉黄芩素(0~100μmol/L)对MMP-2、APN和EGFR-TPK的抑制作用,并进一步的分析了汉黄芩素(0~100μmol/L)对肺癌A549的生长、细胞转移的抑制作用和A549细胞中TIMP-2和TGF-β1表达水平的影响,酶标仪吸光度法检测细胞凋亡蛋白Caspase 3活性。结果:汉黄芩素能够抑制MMP-2、APN和EGFR-TPK的活性(汉黄芩素半抑制浓度IC50分别为7.26±0.33、7.11±0.43和5.32±0.22μmol/L)和诱导A549的凋亡(汉黄芩素半抑制浓度IC50为49.13±4.90μmol/L);随着汉黄芩素作用浓度(1、10、50、100μmol/L)的增加,TGF-β1表达均呈现降低趋势,TIMP-2的表达呈增加趋势,且A549细胞穿过人工基底膜的能力下降而凋亡蛋白Caspase 3活性增加。结论:汉黄芩素通过诱导TIMP-2表达增强和TGF-β1表达的降低,对MMP-2、APN和EGFR-TPK的活性产生抑制作用,对Caspase 3活性产生激活作用,最终对肺癌A549细胞的转移和凋亡起到重要作用。

黄芩素对肥胖模型大鼠减肥作用及机制的研究

目的:探讨黄芩素的减肥作用及其对瘦素表达、β-肾上腺素受体的影响。方法:建立大鼠营养性肥胖模型,观察黄芩素给药4周对肥胖大鼠体重、脂肪湿重、脂肪细胞个数及直径(光镜下)、血清瘦素含量(ELISA法)以及脂肪细胞瘦素表达等指标的影响,同时应用放射配基结合实验观察黄芩素与β-肾上腺素受体及β3肾上腺素受体的结合情况。结果:黄芩素(100 mg/kg)能明显降低肥胖大鼠体重、Lee's指数、脂肪重量和脂体系数以及血清瘦素水平,黄芩素镜下脂肪细胞个数明显增加,细胞直径明显减小,黄芩素(100 mg/kg和50 mg/kg)模型动物脂肪细胞瘦素表达明显降低。黄芩素与β肾上腺素受体结合的K值为80.36nmol/L,与β3肾上腺素受体结合的K值为35.11nmol/L。结论:黄芩素对营养性肥胖大鼠模型具有减肥作用,并对营养饲料诱导的瘦素抵抗状态具有显著的改善作用,黄芩素对β-肾上腺素受体及β3肾上腺素受体均具有一定的亲和力。

竞争型MMP-2抑制剂黄芩素对口腔鳞状癌细胞生长和转移作用的影响

目的：探讨明胶酶（MMP-2）抑制剂黄芩素对口腔鳞状细胞癌细胞的生长和转移抑制机制。方法：采用酶抑制动力学方法,研究了黄芩素（0～150μmol/L）对MMP-2的抑制作用,并进一步的分析了黄芩素（0～200μmol/L）对口腔鳞状细胞癌Tca-8113的生长的抑制作用,Tca-8113细胞转移的抑制作用和Tca-8113细胞中TIMP-2、MMP-2和TGF-β1表达水平的影响。结果：黄芩素能够竞争性的抑制MMP-2的活性和诱导Tca-8113的凋亡（黄芩素半抑制浓度IC50分别为17.26±1.23μmol/L和24.39±0.97μmol/L）;随着黄芩素作用浓度（5、10、50、100μmol/L）的增加,MMP-2和TGF-β1表达均呈现降低趋势,TIMP-2的表达呈增加趋势,且肿瘤细胞穿过人工基底膜的能力下降。结论：黄芩素能够通过抑制MMP-2的活性和增强TIMP-2的表达,诱导了MMP-2和TGF-β1表达的降低,最终对口腔鳞状细胞癌Tca-8113的转移和凋亡起到重要作用。

High sensitivity detection of baicalein by N,S co⁃doped carbon dots and their application in biofluids

In this work,p⁃phenylenediamine and L⁃cysteine were used as raw materials,and water⁃soluble N,S co⁃doped carbon dots(N,S⁃CDs)with excellent performance were prepared through a one⁃step solvothermal method.The morphology and structure of N,S⁃CDs were characterized by transmission electron microscope,X⁃ray diffrac⁃tion,Fourier transform infrared spectroscopy,and X⁃ray photoelectron spectroscopy,and the basic photophysical properties were investigated via UV⁃Vis absorption spectra and fluorescence spectra.Meanwhile,the N,S⁃CDs have excellent luminescence stability with pH,ionic strength,radiation time,and storage time.Experimental results illus⁃trated the present sensor platform exhibited high sensitivity and selectivity in response to baicalein with a detection limit of 85 nmol·L-1.The quenching mechanism is proved to be the inner filter effect.In addition,this sensor can also detect baicalein in biofluids(serum and urine)with good accuracy and reproducibility.

黄芩的生物活性及其在化妆品中的应用

论述了黄芩的资源、主要化学成份、提取方法、生物活性及其在化妆品中的应用等。黄芩中的主要活性成份黄芩苷和黄芩素等具有广泛的生物活性,如抗菌、抗真菌、抗炎、抗过敏、抗氧化、清除自由基、抗紫外线、抗辐射和抑制酪氨酸酶等,可制备防治痤疮、防晒、祛斑美白、抗皮肤老化和防皱等功效性化妆品,防治牙周炎等口腔用品以及外用洗剂等。黄芩素和黄芩苷等黄芩中主要活性成份是集祛斑美白、防晒、抗敏、消炎和抗衰老多效合一的天然高效多功能化妆品添加剂。

Simultaneous determination of four active compounds in Dangguilonghui tablet by high-performance liquid chromatography

A high-performance liquid chromatography （HPLC） method has been developed for the first time to simultaneously quantify the four active ingredients, namely aloha, baicalein, aloe-emodin and wogonin, in Dangguilonghui tablet. The marker compotmds were separated on the Diamonsil C18 column at a flow rate of 1.0 mL/min with a mobile phase of methanol-acetonitrile-0.3% aqueous phosphoric acid （220： 4： 200, v/v/v） and while the detection wavelength was set at 225 nm. The assay was linear over the range of 1.450 - 29.00 ug/mL （r = 0.9992） for aloin, 0.4050 - 8.100ug/mL （r = 0.9994） for baicalein, 0.1100 - 2.200 ug/mL （r = 0.9997） for aloe-emodin, 0.2160 - 4.320 ug/mL （r = 0.9991） for wogonin, respectively. The average sample recoveries at three concentration levels were 100.7% （RSD = 0.88%）, 101.0% （RSD = 0.89%）, 100.0% （RSD = 1.3%） and 100.1% （RSD = 1.1%） for these constituents, respectively. This method is suitable for the quality control of Dangguilonghui tablet.

Metabolites of baicalein 6,7-diacetate in rats

To investigate the metabolites of baicalein 6,7-diacetate in rats. HPLC-DAD and LC/MS^n methods were used to analyze the metabolites in intestinal tract and plasma after oral administration of baicalein 6,7-diacetate to rats. Baicalein 6,7-diacetate was degraded into baicalein 6-monoacetate and baicalein in rat intestinal tract, and four baicalein glucuronides, baicalein 6-0- glucuronide, baicalein 6-methoxyl-7-O-glucuronide, baicalein 6,7-di-O-glucuronide, and baicalein 6-O-glucose-7-O-glucuronide were detected and tentatively identified in rat plasma. This is the first time to report the metabolites of baicalein 6,7-diacetate in rats. Six metabolites were identified in rat intestinal tract and plasma.

Experimental and Theoretical Investigation on Excited State Intramolecular Proton Transfer Coupled Charge Transfer Reaction of Baicalein

The excited state intramolecular proton transfer （ESIPT） coupled charge transfer of baicalein has been investigated using steady-state spectroscopic experiment and quantum chemistry calculations. The absence of the absorption peak from S1 excited state both in the experi-mental and calculated absorption spectra indicates that S1 is a dark state. The dark excited state S1 results in the very weak fluorescence of solid baicalein in the experiment. The fron- tier molecular orbital and the charge difference densities of baicalein show clearly that the S1 state is a charge-transfer state whereas the S2 state is a locally excited state. The only one stationary point on the potential energy profile of excited state suggests that the ESIPT reaction of baicalein is a barrierless process.

Separation of baicalin from Scutellaria Baicalensis Georgi with polyamide

Separation of baicalin from Scutellaria Baicalensis Georgi with polyamide was studied. The adsorption isotherm, kinetic equation and desorption law were investigated by static and dynamic adsorption methods. The results show that the kinetic behavior is mainly controlled by the liquid film diffusion process and obeys the Boyd film diffusion equation. Equilibrium data for the adsorption of baicalin are correlated with Freundlich isotherm equation, i.e. q=3.8ce2.057, suggesting that the relative capacity of polyamide to baicalin is somewhat small. The desorption results indicate that the baicalin with mass fraction of 33.86% and the least impurities can be obtained by chromatography using 60% ethanol as the eluant at room temperature.

A New Flavone C glycoside from Scutellaria baicalensis

A new flavone C glycoside, chrysin 8 C β D glucoside (1) and a known phenethyl alcohol glycoside, acteoside (2), were isolated for the first time from the roots of Scutellaria baicalensis (Labiatae).

Different Treatment Methods on Seed Germination Seedlings

[Objective] This study was conducted to investigate the effects of different gibberellic acid and hydrogen peroxide concentrations and different soaking time on the germination potential and germination rate of Scutellaria baicalensis, so as to improve the breeding efficiency of S. baicalensis. [Method] Dry seeds were divided into 3 groups, each containing 50 grains, which were weighed and put in a culture dish and soaked with distilled water covering the surface of seeds at room temperature. The seed weight was measured by gravimetric method once every 30 min from 30 rain after soaking, until the weight of the seeds no longer changed. [Result] With the increase of the concentration of hydrogen peroxide, the germination rate of S. baicalensis was improved gradually, and reached 32.0% under the concentration of 1.0% at the soaking time of 24 h; and with the increase of the concentration of gibberellic acid, the germination rate of S. baicalensis was improved gradually, and reached 64.0% under the concentration of 600 mg/L at the soaking time of 24 h. [Conclusion] The constant temperature at 25 ℃ in the incubator could also improve the seed germination rate; and the germination rate achieved by treating the seeds with gibbereUic acid was remarkably higher than that achieved by treating the seeds with hydrogen peroxide.

Studies on the Constituents of Aerial Parts of Scutellaria planipes

首次从木根黄芩地上部分分离出１３个化合物。经理化常数和光谱分析分别鉴定为异甘草甙元（ｉｓｏｌｉｑｕｉｒｉｔｉｇｅｎｉｎ），蓟黄素（ｃｉｒｓｉｍａｒｉｔｉｎ），汉黄芩素（ｗｏｇｏｎｉｎ），芹菜素（ａｐｉｇｅｎｉｎ），粘毛黄芩素Ⅲ（ｖｉｓｃｉｄｕｌｉｎⅢ），紫丁香甙（ｓｙｒｉｎｇｉｎ），白杨素６ＣβＤ葡萄糖８ＣαＬ阿拉伯糖甙（６ＣβＤｇｌｕｃｏｓｙｌ８ＣαＬａｒａｂｉｎｏｓｙｌｃｈｒｙｓｉｎ），白杨素６ＣαＬ阿拉伯糖８ＣβＤ葡萄糖甙（６ＣαＬａｒａｂｉｎｏｓｙｌ８ＣβＤｇｌｕｃｏｓｙｌｃｈｒｙｓｉｎ），黄芩甙（ｂａｉｃａｌｉｎ），汉黄芩甙（ｗｏｇｏｎｉｎ７ＯβＤｇｌｕｃｕｒｏｎｉｄｅ），白杨素７ＯβＤ葡萄糖醛酸甙（ｃｈｒｙｓｉｎ７ＯβＤｇｌｕｃｕｒｏｎｉｄｅ）和两个苯乙醇甙（ａｃｔｅｏｓｉｄｅ和ｍａｒｔｙｎｏｓｉｄｅ）。其中异甘草甙元，蓟黄素和紫丁香甙均为首次从黄芩属植物中分离得到。

Antifungal susceptibility analysis of berberine,baicalin,eugenol and curcumin on Candida albicans

Objective:To analyze the antifungal effects of Chinese herb monomers,i.e.berberine,baicalin,eugenol and curcumin,on Candida albicans.Methods:After Candida albicans strain Y01-09 was incubated for 48 h in YEPD broth which contained different concentrations of Chinese herb components,the cell cycle,fluorescent intensity and the size of cell volume were detected by flow cytometry.Results:The 4 Chinese herb monomers could affect the cell cycle of Candida albicans in different ranges.The ratio of cells in S-G2-M period decreased as the agents concentration increased,indicating that the cell division was inhibited.The fluorescent intensity of Candida albicans cells became weaker after being incubated,which reflected the loss of DNA fragments.The higher the concentration was,the weaker the fluorescent intensity became.The cell size,cell diopter and particle size changed much as the agents concentration increased.Conclusion:Chinese herb monomers play the antifungal role in inhibiting cell division.FCM could be used to determine the susceptibility of antifungal agents.

Increased life-span of Drosophila melanogaster by Flavonoids from Scutellariabaicalensis Georgi induced by hydrogen peroxide

Objective： To explore the effects and mechanisms of flavonoids from Scutellaria baicalensis Georgi in hydrogenperoxide （H2O2） induced Drosophila melanogaster model. Method： H2O2 induced Drosophila melanogaster agingmodel was used to study the effects of flavonoids from Scutellaria baicalensis Georgi. Secondly, the effects ofbaicalein and wogonin on mRNA expressions were examined. Results： Our results found that baicalein andwogonin could extend the life-spans of H2O2 induced Drosophila melanogaster aging model via increased mRNAexpressions of superoxide dismutase catalase and rosy. Conclusion： Baicalein and wogonin could extend life-spansof H2O2 induced Drosophila melanogaster aging model through increasing mRNA expressions of superoxidedismutase, catalase and rosy.

The effect of baicalein on the expression of SATB1 in MDA-MB-231 cells

Baicalein had been proved to have anti-cancer activity in vitro and in vivo, including the inhibition of malignant proliferation, migration, adhesion and invasion of many kinds of cancer cells. The special AT-rich sequence binding protein 1 （SATB1） is a tissue-specific expression of nuclear matrix-binding protein and is reported to be a breast cancer ＂gene group organizer＂. Previous studies have shown that SATB1 is involved in the growth, metastasis and prognosis of breast cancer. The present study was aimed to investigate whether baicalein inhibits the proliferation and migration of MDA-MB-231 human breast cancer cells through down-regulation of the SATB1 expression. Methods： MDA-MB-231 cells were treated for 24 h, 48 h and 72 h with various concentrations of baicalein （0, 5, 10, 20, 40 and 80 pM） respectively. Then, the proliferation and migration of MDA-MB-231 cells following treatment with baicalein were determined using colorimetric 3-（4, 5-dimethylthia- zol-2-yl） 2, 5-diphenyltetrazolium bromide （MTT） and wound healing assays. Thereafter, western blot analysis was performed to detect the changes of SATB1 protein expression in MDA-MB-231 cells. Results： Along with the prolongation of time and increase of drug concentration, inhibitory effect of baicalein on proliferation and migration of MDA-MB-231 cells gradually in- creased, in a time.- and dose- dependent manner （P 〈 0.05）. Meanwhile, after treated with baicalein in different concentrations for 48 h, the level of SATB1 protein expression of MDA-MB-231 cells decreased obviously, in a dose-dependent manner （P 〈 0.05）. Conclusion： Baicalein inhibits breast cancer cell proliferation and suppresses its invasion and metastasis by reducing cell migration possibly by down-regulation of the SATB1 protein expression, indicating that baicalein is a potential therapeutic agent for human breast cancer.

Antiarthritic Activity of Apigenin, Scutellarein and Pectinolinergenin Glycosides from Clerodendron inerme

The present study was aimed to investigate the anti-arthritic activity of alcoholic extract ofClerodendron inerme which was shown to inhibit inflammatory process. Arthritis was induced in rats by injecting rats intradermally with Freund＇s adjuvant 0.01%-0.1% in paraffin oil. They were grouped and received 10 mg/100 gm b.wt. of alcoholic fraction of C. inerme, 1.5 mg/100 gm b.wt. hydrocortisone and saline, orally, for 15 days. The arthritic scores, radiological scores, body weights, liver glycogen, serum mucoproteins were monitored for 49 days. Wilcoxan ranking methods and Student＇s t Test were used to determine the significance changes between control and treated groups. Adjuvant-arthritic rats showed a significant decrease in body weight, liver glycogen, increased serum mucoprotein and severity of periostitis and other degenerative changes m the bone. But treatment with the alcoholic fraction （apigenin, scutallarin and pectinolinergenin） of C. inerme exhibited a very good relief from the arthritic conditions by increasing the body weight, increased liver glycogen content and registered little degenerative changes in bones, suggesting long term anti-arthritic activity when compared to the control group. The anti-arthritic activity ofC. inetme alcoholic fraction was remarkable in this study. However, further studies are required to elucidate its full anti-arthritic potential.

Metabolite identification and the development of a simultaneous quantification method for wogonin and wogonin-7-O-glucuronide:application to a distribution study in mice liver

Wogonin possesses potent inhibitory activities against cancer cell growth in vitro and in vivo and has attractive safety profiles.A highly sensitive liquid chromatography coupled with tandem mass spectrometry method was developed for the identification of major metabolites in mice liver after intravenous administration of wogonin.Five metabolites were identified and biotransformation pathways were elucidated as well.Furthermore,a method was developed and validated for the simultaneous quantitatively determination of wogonin and wogonin-7-O-glucuronide in mice liver.After liquid-liquid extraction by ethyl acetate,the analytes were separated on a C_（18）column with a mobile phase of methanol-10 mM ammonium acetate water （80：20,v/v）.The detection was operated with negative selected reaction monitoring mode using electrospray ionization technique. The linear response range was 0.2-40μg/g for both wogonin and wogonin-7-O-glucuronide in mice liver.The developed quantification method was suitable for distribution study after intravenous infusion of wogonin injection in animals.

Identification of the metabolites of baicalein in rat serum

Baicalein,the aglycon of baicalin,is one of the main bioactive flavones in Scutellaria baicalensis Georsi.Five metabolites（M1-M5）and the parent drug（MO）were identified in rat serum after oral administration of baicalein at a dose of 200 mg/kg using the HPLC-DAD and LC-MS/MS methods.Based on the results,the possible metabolic pathway of baicalein in rat serum was proposed.

Baicalein protects against the development of angiotensin II-induced abdominal aortic aneurysms by blocking JNK and p38 MAPK signaling

An abdominal aortic aneurysm(AAA) is a permanent, localized dilatation of the abdominal aorta. In western countries, the morbidity of AAA is approximately 8%. Currently, pharmacotherapies for AAA are limited. Here, we demonstrate that baicalein(BAI), the main component of the Chinese traditional drug "Huang Qin", attenuates the incidence and severity of AAA in Apoe儃/儃 mice infused with angiotensin II(AngII). Mechanically, BAI treatment decreases AngII-induced reactive oxygen species(ROS) production in the aortic wall. Moreover, BAI inhibits inflammatory cell accumulation in the aortas of mice infused with AngII. It also inhibits AngII-induced activation of matrix metalloproteinase 2(MMP-2) and MMP-9 to maintain elastin content in vivo. In addition, it blocks AngII cascade by downregulating angiotensin type 1 receptor(AT1R) and inhibiting mitogen-activated protein kinases(MAPKs). Taken together, our findings show that BAI is an effective agent for AAA prevention.

Autotetraploidy induced in Nianmaohuangqin (Radix Scutellariae viscidulae) with colchicine in vitro

OBJECTIVE： To establish and optimize the propaga- tion of Nianmaohuangqin （Radix Scutellariae Viscid- ulae） and induce and characterize polyploidy of Nianmaohuangqin （Radix Scutellariae Viscidulae）. METHODS： Buds from germinating seed-derived explants were induced by tissue culture. With an or- thogonal test, different concentrations of 6-benzyl- aminopurine （BAP）, indole-3-acetic acid （IAA） and kinetin （KT） were used to determine the optimal concentrations for the propagation of Nianmaohuangqin （Radix Scutellariae Viscidulae）. The differ- ent concentrations of IAA and rooting powder （ABT） were used to induce rooting. A 0.3% w/v col- chicine solution was used to induce polyploidy and the induced buds was identified by root-tip chromosome determination and stomatal apparatus ob- servation. RESULTS： A large number of buds could be in- duced directly from epicotyl and hypocotyl ex- plants on Murashige and Skoog （MS） （Murashige and Skoog 1962） medium supplemented with 1.1-1.3 mg/L BAP and 0.2 mg/L IAA. Root induction and development could be observed within 20 days of inoculation on 1/2 MS medium supplemented with 0.2 mg/L IAA and 0.1 mg/L ABTo Furthermore, 27 lines of autotetraploid individuals were ob- tained with a plantlet chromosome number of 2n= 4x=36. CONCLUSION： Autotetraploid lines could be ob- tained through induction with colchicine in vitro, proving that this method might be used for plant selection and breeding.