紫草素对肺腺癌A549细胞的生长抑制作用及其机制研究

目的观察紫草素抑制人肺腺癌A549细胞的增殖及其机制。方法用不同浓度紫草素处理A549细胞,CCK-8方法检测紫草素对A549细胞生长增殖的抑制作用;细胞周期检测试剂盒测定细胞周期的变化;Western blot法检测磷酸化Akt蛋白（pAkt）的表达。结果紫草素能够抑制人肺腺癌A549细胞的增殖;诱导A549细胞凋亡;阻滞细胞G1期向S期的发展。pAkt蛋白表达量在紫草素处理48 h后明显减少。结论紫草素可以抑制肺腺癌A549细胞的增殖,诱导凋亡,机制可能与紫草素抑制PI3K/Akt信号途径有关。     

β-羟基异戊酰紫草素联合顺铂对SKOV3细胞活力的影响

目的观察β-羟基异戊酰紫草素（β-HIVS）联合顺铂对人卵巢癌细胞株SKOV_3活力的影响,并探讨其可能的发生机制。方法设立空白对照组及6个不同浓度β-HIVS组（2~30μmol/L）,应用四甲基偶氮唑蓝（MTT）比色法测定不同浓度β-HIVS对SKOV_3细胞活力的影响。设立顺铂单用组（10、20、40μmol/L）和选择〈IC_（50）的β-HIVS 3个浓度（0.25、1、2.5μmol/L）协同顺铂组,应用MTT比色法测定β-HIVS协同顺铂对SKOV_3细胞活力的影响。应用Western blot法检测不同浓度β-HIVS作用后BcI-2、Bax蛋白表达。结果不同浓度β-HIVS（2～30μmol/L）均可抑制SKOV_3细胞活力,并呈一定的浓度依赖性,测得半数抑制率IC_（50）为7.37μmol/L,与空白对照组比较,差异均有统计学意义（P〈0.05）。β-HIVS（1、2.5μmol/L）与顺铂联合应用,与顺铂单用组比较,差异亦有统计学意义（P〈0.05,P〈0.01）。协同作用对β-HIVS呈现浓度依赖性。Western bIot检测结果表明：不同浓度β-HIVS（5、7.5、10μmol/L）可明显上调SKOV_3细胞中Bax蛋白表达水平,同时抑制Bcl-2蛋白表达,与对照组比较,差异有统计学意义（P〈0.01）。结论β-HIVS对人卵巢癌细胞株SKOV_3体外活力有明显的抑制作用,且呈浓度依赖性。在一定浓度范围内,β-HIVS对顺铂具有协同增敏作用,且随β-HIVS浓度的增高,这种协同作用表现的更为显著。协同作用可能通过β-HIVS上调Bax蛋白表达和抑制Bcl-2蛋白表达,从而加速诱导SKOV_3凋亡。     

高效液相色谱流通池法测定紫草中假紫草素的含量

高效液相色谱法测定紫草提取物中的假紫草素时,出现20个峰,需时75min,主峰保留时间55min,常规方法制备工作曲线很费时。本文报道一种新的工作曲线法——流通池法的原理和方法,测定结果与常规方法一致,可以节省时间。     

紫草素对子宫内膜异位症大鼠异位病灶的影响

目的探究紫草素对子宫内膜异位症(EMs)大鼠异位病灶及子宫内膜组织基质细胞衍生因子-1(SDF-1)、基质细胞衍生因子受体-4(CXCR4)表达的影响。方法雌性SD大鼠120只,随机分为对照组、模型组、实验组及阳性对照组,每组30只。除对照组外,其余3组大鼠均采用自体移植法建立EMs大鼠模型。造模成功后,实验组大鼠灌胃紫草素(10 mg·kg-1·d-1),阳性对照组大鼠灌胃孕三烯酮(0.23 g·kg-1·d-1),对照组与模型组灌胃等量生理盐水,连续干预15 d。干预结束后,测定异位病灶体积;采用苏木精-伊红(HE)染色观察大鼠子宫内膜组织形态学变化;采用酶联免疫吸附(ELISA)法检测血清卵泡刺激素(FSH)、黄体生成激素(LH)、雌激素(E2)、孕激素(P)水平;采用免疫组织化学法检测子宫内膜组织SDF-1、CXCR4蛋白表达。结果对照组、模型组、实验组及阳性对照组大鼠异位病灶体积分别为0,(126.62±79.34),(28.36±24.12),(16.38±14.51)mm3;子宫内膜组织SDF-1蛋白阳性细胞百分比分别为(19.23±2.86)%,(84.38±3.11)%,(62.42±3.73)%,(41.12±4.25)%;CXCR4蛋白阳性细胞百分比分别为(28.06±2.54)%,(78.68±3.23)%,(56.45±2.93)%,(43.13±3.26)%。与模型组比较,实验组大鼠异位病灶的体积缩小,且阳性对照组小于实验组;模型组大鼠血清FSH、LH、P水平较对照组显著降低,与模型组比较,实验组升高,且阳性对照组高于实验组;血清E2水平及子宫内膜组织SDF-1、CXCR4蛋白阳性细胞百分比均升高,与模型组比较,实验组降低,且阳性对照组低于实验组(均P<0.05)。结论紫草素可抑制EMs大鼠异位病灶的发展,同时抑制E2的异常分泌,提高FSH、LH、P水平,降低子宫内膜组织中SDF-1、CXCR4蛋白表达,进而抑制异位内膜的生长,达到治疗EMs的目的。     

Comparison of some secondary metabolite content in the seventeen species of the Boraginaceae family

Context: The Boraginaceae family comprises plants that have important therapeutic and cosmetic applications. Their pharmacological effect is related to the presence of naphthaquinones, flavonoids, terpenoids, phenols, or purine derivative – allantoin.

Objective: In the present study, comparison of some secondary metabolite content and phytochemical relationship between 17 species of the Boraginaceae family were analyzed.

Materials and methods: High performance capillary electrophoresis (HPCE) was used to perform a chemometric analysis in the following Boraginaceae species: Anchusa azurea Mill., Anchusa undulata L., Borago officinalis L., Buglossoides purpurocaerulea (L.) I.M. Johnst., Cerinthe minor L., Cynoglossum creticum Mill, Echium italicum L., Echium russicum J.F. Gmel., Echium vulgare L., Lindelofia macrostyla (Bunge) Popov (syn. Lindelofia anchusoides (Lindl.) Lehm.), Lithospermum officinale L., Nonea lutea (Desr.) DC., Omphalodes verna Moench (syn. Cynoglossum omphaloides L.), Pulmonaria mollis Wulfen ex Hornem., Pulmonaria obscura Dumort., Symphytum cordatum Waldst. & Kit ex Willd., and Symphytum officinale L.

Results: Six active compounds in shoot extracts (allantoin, p-hydroxybenzoic acid, rutin, hydrocaffeic acid, rosmarinic acid, and chlorogenic acid) and four compounds in root extracts (allantoin, hydrocaffeic acid, rosmarinic acid, and shikonin) were identified. The presence and abundance of these compounds were used for the characterization of the species and for revealing their phytochemical similarity and differentiation.

Discussion and conclusion: The present study provides the first comprehensive report of the extraction and quantification of several compounds in Boraginaceae species (some of them for the first time). Among the 17 species studied, species with potentially high pharmacological activity were recognized.     

Cellular pharmacology studies of shikonin derivatives

The naphthoquinone pigment, shikonin, isolated from Lithospermum erythrorhizon Sieb. et Zucc.(Boraginaceae) and its derivatives are the active components isolated from the Chinese herbal therapeutic, Zicao. Historically, Zicao root extracts have been used to treat macular eruption, measles, sore-throat, carbuncles and burns. Multiple pharmacological actions have been attributed to shikonin, e.g. antiinflammatory, antigonadotropic and anti-HIV-1 activity. In this review, several therapeutic applications of shikonin will be summarized including its pleiotropic, antiinflammatory and antitumour effects. Widely diverse and sometimes conflicting activities have been attributed to shikonin, e.g. wound healing, enhanced granuloma formation, suppression of local acute inflammatory reactions, inhibition of angiogenesis, inhibition of select chemokine ligands, inhibition of DNA topoisomerase activity, inhibition of platelet activation and antimicrobial activity. Comparison of the various reported mechanisms of action for shikonin lead us to hypothesize that shikonin is an effective inhibitor of protein-protein interaction with multiple targets in both the intracellular and extracellular compartments. This general inhibitory effect can account for the broad spectrum of shikonin biological and pharmacological activities.     

紫草素通过PI3K/Akt通路促进人乳腺癌MCF-7细胞自噬

目的研究紫草素(Shikonin)对人乳腺癌MCF-7细胞自噬的影响及其作用机制。方法 CCK-8法检测紫草素处理人乳腺癌MCF-7细胞24、48 h细胞的存活率,Western blot方法检测紫草素处理24 h和48 h时LC3、p62、PI3K、Akt、p-PI3K、p-Akt蛋白水平。结果 1μmol.L-1紫草素处理细胞48 h以及2.5、5μmol.L-1紫草素处理MCF-7细胞24 h和48 h时,MCF-7细胞的活力受到明显抑制,LC3-Ⅱ/LC3-Ⅰ值增加,p62表达减少,总PI3K、Akt、p-PI3K、p-Akt均减少。结论紫草素促进乳腺癌MCF-7细胞自噬,其作用机制可能与PI3K/Akt通路受到抑制有关。     

Design,Synthesis and Biological Evaluation of Cinnamic Acyl Shikonin Derivatives

Inducing apoptosis is an important and promising therapeutic approach to overcome cancer. Here, we described a series of novel synthesized compounds, cinnamic acyl shikonin derivatives ( 1b – 19b ), which were synthesized starting from shikonin and cinnamic acids, which exhibit anticancer activity via inducing apoptosis in vitro. Our flow cytometry results showed that compound 8b ((E)‐1‐(5,8‐dihydroxy‐1,4‐dioxo‐1,4‐dihydronaphthalen‐2‐yl)‐4‐methylpent ‐3‐enyl‐3‐(3‐(trifluoromethyl) phenyl)acrylate) (IC₅₀ = 0.69, 0.65, 1.62 μm for human SW872‐s, A875 and A549 cell lines, respectively) exhibited conspicuous anticancer activities and has low cell toxicity in vitro. Therefore, we considered that compound 8b is potentially to be a candidate of anticancer agent. The proliferation inhibitory effect of compound 8b was associated with its apoptosis‐inducing effect by activating caspase‐3, caspase‐7, caspase‐9, and PARP. When the level of cleaved caspase‐3, cleaved caspase‐7, cleaved caspase‐9, and cleaved PARP are rise, apoptosis of cancer cells will be induced.     

紫草蒽醌类成分的提取及其复合物栓的抗炎活性研究

目的：基于预防和治疗老年性阴道炎及妇科炎症等疾病,从开发绿色天然中药制剂出发,优选紫草复合物栓最佳提取工艺,并以抗炎活性为指标,优化紫草提取物在复合物中配比。方法：采用正交试验设计方法,以出膏率、总萘醌含量及左旋紫草素含量为综合指标进行工艺优选,二甲苯致炎实验优化处方配比。结果：紫草的最佳提取工艺为：浸泡4小时后,8倍量90%的乙醇渗漉。当紫草占处方总量的50%时,其抗炎活性最强。结论：优选的提取工艺,条件稳定,方法简单,可操作性强且成本低,可用于紫草复合物栓的制备。以活性为指标的处方筛选,为紫草复合物栓的有效性开发提供重要依据。