Effects of tanshinone Ⅱ-A sullfonate on adhesion molecule expression of endothelial cells and platelets in vitro

探讨丹参酮ⅡＡ磺酸钠（Ｔａｎ）对培养人脐静脉内皮细胞（ＨＵＶＥＣ）和人血小板表达粘附分子的影响．方法：用流动血细胞计数仪测定肿瘤坏死因子（ＴＮＦα）诱导人脐静脉内皮细胞ＩＣＡＭ１和凝血酶诱导人血小板Ｐ选择素的表达．结果：ＨＵＶＥＣ经ＴＮＦα处理后，明显增加细胞表面ＩＣＡＭ１的表达，增加ＨＬ６０细胞粘附到内皮细胞表面达加入细胞总数的３０％±６％（对照组为４６％±０７％）．在ＴＮＦα处理前，用Ｔａｎ（２５－２００μｍｏｌ·Ｌ－１）与ＨＵＶＥＣ共孵育，则Ｔａｎ剂量依赖性地抑制ＴＮＦα的作用．Ｔａｎ（２５－２００μｍｏｌ·Ｌ－１）与人血小板孵育后，可剂量依赖性地抑制凝血酶诱导人血小板表面Ｐｓｅｌｅｃｔｉｎ的表达．结论：Ｔａｎ可抑制内皮细胞和血小板表达粘附分子．     

丹参酮ⅡA磺酸钠注射液对非ST段抬高心肌梗死患者血浆组织型纤溶酶原激活物抑制物的影响

目的 观察非ST段抬高心肌梗死患者血浆组织型纤溶酶原激活物抑制物(PAI-1)的变化和丹参酮Ⅱ A磺酸钠注射液对其的影响.方法 将非ST段抬高心肌梗死患者136例随机分为治疗组72例和对照组64例.2组均予西药常规治疗,治疗组另加用丹参酮Ⅱ A磺酸钠注射液治疗.2组均14d为1个疗程.2组分别于治疗前、治疗后14d测定血浆PAI-1浓度,并观察其严重心律失常、心绞痛、心力衰竭的发生情况.结果 治疗后2组PAI-1水平均下降(P＜0.05),但治疗组下降更明显(P＜0.05).治疗组严重心律失常、心绞痛、心力衰竭的发生率均显著低于对照组,差异均有统计学意义(P＜0.05).结论 丹参酮Ⅱ A磺酸钠注射液辅治非ST段抬高心肌梗死疗效确切,并能降低其严重心律失常、心绞痛、心力衰竭的发生率.     

丹参酮ⅡA磺酸钠注射液对非ST段抬高心肌梗死患者血浆组织型纤溶酶原激活物抑制物的影响

目的观察非ST段抬高心肌梗死患者血浆组织型纤溶酶原激活物抑制物（PAI-1）的变化和丹参酮ⅡA磺酸钠注射液对其的影响。方法将非ST段抬高心肌梗死患者136例随机分为治疗组72例和对照组64例。2组均予西药常规治疗,治疗组另加用丹参酮ⅡA磺酸钠注射液治疗。2组均14d为1个疗程。2组分别于治疗前、治疗后14d测定血浆PAI-1浓度,并观察其严重心律失常、心绞痛、心力衰竭的发生情况。结果治疗后2组PAI-1水平均下降（P〈0.05）,但治疗组下降更明显（P〈0.05）。治疗组严重心律失常、心绞痛、心力衰竭的发生率均显著低于对照组,差异均有统计学意义（P〈0.05）。结论丹参酮ⅡA磺酸钠注射液辅治非ST段抬高心肌梗死疗效确切,并能降低其严重心律失常、心绞痛、心力衰竭的发生率。     

沉默LIMK1促进DADS抑制SW480细胞迁移与侵袭

目的探讨LIMK1沉默对DADS抑制SW480细胞迁移与侵袭的影响。方法 RNA干扰技术建立稳定LIMK1-miRNA/SW480细胞株;免疫组化和Western blot检测DADS对沉默LIMK1结肠癌SW480细胞LIMK1和磷酸化LIMK1蛋白表达的影响。划痕实验和侵袭实验检测LIMK1 RNA沉默与DADS对SW480细胞迁移与侵袭的影响。结果 RT-PCR与Western blot显示,LIMK1-miR/SW480细胞LIMK1mNRA与蛋白表达明显下调,表明成功构建稳定沉默LIMK1基因的SW480细胞株。免疫组化和Western blot显示,45 mg.L-1DADS处理和沉默LIMK1组较未转染组与空载体组SW480细胞LIMK1蛋白和磷酸化LIMK1明显下调(P<0.05)。划痕实验和侵袭实验发现,沉默LIMK1或DADS处理SW480细胞的迁移与侵袭能力较未转染组与空载体组明显抑制(P<0.05)。而DADS处理沉默组抑制SW480细胞迁移和侵袭能力更为明显(P<0.05)。结论沉默LIMK1基因可抑制SW480细胞迁移与侵袭,增强DADS抑制SW480细胞迁移与侵袭作用。     

Apoptosis inducing effects of 6-Methoxydihydrosanguinarine in HT29 colon carcinoma cells

6-methoxydihydrosanguinarine (6ME), a benzophenanthridine alkaloid derived from the methanol extracts of Hylomecon hylomeconoides, showed a dose-dependent effect at 1-10 microM on causing apoptotic cell death in HT29 colon carcinoma cells (IC50 = 5.0+/-0.2 microM). Treatment of HT-29 cells with 6ME resulted in the formation of internucleosomal DNA fragmentation. Treatment of the cells with 6ME caused activation of caspase-3, -8 and 9 protease and subsequent proteolytic cleavage of poly(ADP-ribose)polymerase. 6ME increased the expression of p53 and Bax and decreased the expression of Bid. These results indicate that p53 and proapoptotic Bcl-2 family proteins might participate in the antiproliferative activity of 6ME in HT29 cells.     

姜黄素与5-氟尿嘧啶联用对人结肠癌HT-29细胞增殖的影响

目的　体外观察 5 氟尿嘧啶与姜黄素联用对人结肠癌HT 2 9细胞增殖的相互作用。方法　采用MTT法观察不同浓度姜黄素、5 氟尿嘧啶单独或联合应用对HT 2 9细胞生长的抑制作用 ,并利用中效原理判定两药合用的效果。结果　两种药物单独应用时 ,对HT 2 9细胞的抑制作用呈明显的剂量效应关系 ;对于HT 2 9细胞 ,姜黄素的中位抑制浓度为(32± 11) μmol·L- 1,5 氟尿嘧啶的中位抑制浓度为(10 4 0± 4 5 6 ) μmol·L- 1。应用中效原理分析显示 ,两药在联合应用时为协同效应 ,并且在不同的药物浓度配比下呈相同的趋势。增大姜黄素的用量可在更宽的效应范围内获得两种药物的协同效应。结论姜黄素与 5 氟尿嘧啶在联合应用时的相互作用为协同效应 ,本结果对于结肠癌的治疗具有一定的临床意义。     

白头翁皂苷A3通过M2型巨噬细胞提高人结肠癌SW480细胞对5-FU的化疗敏感性

目的:观察白头翁皂苷A3(A3)通过干预巨噬细胞极化增强人结肠癌SW480细胞对5-氟尿嘧啶(5-FU)化疗的敏感性,并探讨其作用机制。方法:采用CCK8法检测A3对人单核THP-1细胞存活率的影响,确定A3的安全作用浓度;采用佛波酯(PMA)诱导THP-1细胞分化为M0型巨噬细胞,将M0型巨噬细胞分为M0对照组,M2模型组,A3低、中、高剂量干预组。其中M2模型组给予20 μg·L^-1IL-4与20 μg·L^-1IL-13进行造模,A3干预组在造模的同时给予低、中、高剂量(50,75,100 mg·L^-1)A3进行干预。采用流式细胞术检测M2型巨噬细胞表面抗原CD206的表达;采用RT-PCR法检测M2型巨噬细胞极化因子CD206、IL-10、CCL22、CCL18 mRNA的表达;采用Western blot法检测细胞内p-STAT6、CD206、IL-10、CCL22蛋白的表达。A3干预巨噬细胞M2型极化后的细胞上清液作为条件培养基用于考察SW480细胞对5-FU的化疗敏感性,采用CCK8法检测SW480细胞的存活率;采用Annexin-FITC/PI双染法及Western blot法检测SW480细胞凋亡水平,以及Bax、Cleaved Caspase-3蛋白的表达。结果:A3在50,75,100 mg·L^-1浓度时对THP-1细胞存活率没有显著影响。A3呈剂量依赖性减少CD206阳性细胞表达比例,下调p-STAT6、CD206、IL-10、CCL22、CCL18基因与蛋白的表达(P<0.05,P<0.01)。A3干预巨噬细胞M2型极化后的条件培养基显著提高了SW480细胞对5-FU的敏感性,表现为SW480细胞的存活率下降、凋亡率升高,以及促凋亡蛋白Bax、Cleaved Caspase-3表达上调(P<0.05)。结论:A3能够减弱M2型巨噬细胞对SW480细胞凋亡的抑制作用从而提高SW480细胞对5-FU的化疗敏感性,作用机制可能与A3调控STAT6信号通路抑制巨噬细胞M2型极化有关。     

人结肠癌SW480细胞裸鼠皮下移植瘤模型的建立

目的建立人结肠癌SW480细胞移植瘤模型,研究其形态学及生物学特性。方法将处于对数生长期的人结肠癌SW480细胞接种于10只裸鼠右侧背部皮下,待瘤体生长至直径1～2 cm时处死裸鼠,取出肿瘤组织,将其磨碎并与培养基混合、过滤,滤液与基质胶混合,再将基质胶细胞悬液接种于30只裸鼠右侧背部皮下7,d后处死裸鼠,观察肿瘤大体特征,光镜及电镜下观察肿瘤病理学特征。结果接种SW480细胞的裸鼠于接种后4周有3只成瘤。接种基质胶细胞悬液的裸鼠在接种后7 d有28只成瘤,瘤体形状规则,肿瘤组织呈现典型癌变特征。结论本实验成功建立了人结肠癌裸鼠皮下移植瘤模型,该模型成瘤率高,瘤体形状规则,病理学改变与临床非常相似,为研究人结肠癌干预措施提供了较为理想的动物模型。     

Inhibitory effects of Leucaena leucocephala on the metastasis and invasion of human oral cancer cells

Oral cancer is one of the most common cancers worldwide, and metastasis is recognized as a major factor causing its low survival rate. The inhibition of metastasis progress and the improvement of the survival rate for oral cancer are critical research objectives. Leucaena leucocephala from the mimosa branch Leucaena genus is native to Central and South America and has been used as a traditional remedy for treating various disorders. Previous studies have demonstrated antioxidant, anti‐inflammatory as well as anticancer properties of L. leucocephala plant materials. However, the molecular mechanism underlying the anticancer effect induced by L. leucocephala remains unclear. In this study, we investigated the effect of L. leucocephala extract (LLE) on SCC‐9 and SAS oral cancer cells and examined the potential inhibitory mechanisms involved. The results indicated that LLE attenuated the migration and invasion abilities of both SCC‐9 and SAS cells by reducing the activity and protein expression of matrix metalloproteinases‐2 (MMP‐2). Regarding mitogen‐activated protein kinase (MAPK) pathways, the phosphorylation of ERK1/2 and p38 exhibited a significant inhibitory effect in the presence of LLE. The application of ERK inhibitor and p38 inhibitor confirmed that both signalling transduction pathways were involved in the inhibition of cell metastasis. These data indicate that L. leucocephala could be a potent therapeutic agent for the prevention and treatment of oral cancer and a prominent plant source for anticancer research in the future.     

Crude extract of Rheum palmatum L inhibits migration and invasion of LS1034 human colon cancer cells acts through the inhibition of matrix metalloproteinase‐2/‐9 by MAPK signaling

Crude extract of Rheum palmatum L. (CERP) has been used to treat different diseases in the Chinese population for decades. In this study, we investigated the anti‐metastasis effects of CERP on LS1034 human colorectal cancer cells in vitro and examined potential mechanisms of its effects. CERP significantly inhibited cell migration and invasion of LS1034 cells. We also found that CERP inhibited protein levels of matrix metalloproteinases‐2 (MMP‐2) and matrix metalloproteinases‐9 (MMP‐9), and cytosolic NF‐kB p65, RHO A, ROCK 1. Furthermore, we found CERP inhibited protein levels of GRB2, SOS1, MKK7, FAK, Rho A, ROCK 1, VEGF, PKC, AKT, phosphor‐AKT (Thr308), Cyclin D, iNOS, COX2, NF‐kB p65, p‐ERK1/2, p‐JNK1/2, p‐p38, p‐c‐jun, MMP‐2, MMP‐9, MMP‐1, MMP‐7, MMP‐10, UPA and increased the protein level of Ras in LS1034 cells. In conclusion, our results suggest that CERP may be used as a novel anti‐metastasis agent for the treatment of human colon cancer cells. © 2014 Wiley Periodicals, Inc. Environ Toxicol 30: 852–863, 2015.     

Cytotoxic and anti-inflammatory effects of onion peel extract on lipopolysaccharide stimulated human colon carcinoma cells

The present study investigated the cytotoxic activity of ethanol extract of onion peel (OPE) in HT-29 human colon carcinoma cells. High-performance liquid chromatography (HPLC) analysis was performed to determine the amounts of phenolic acids and flavonoids in OPE. In addition, the influence of OPE on antioxidant- and inflammation-associated gene expression was also determined in a model of lipopolysaccharide (LPS)-stimulated HT-29 cells. HPLC analysis showed that OPE contained well-known antioxidant compounds, including p-coumaric acid, vanillic acid, epicatechin, and morin. After incubation with OPE, HT-29 cells showed either a loss of normal nuclear architecture or detachability from each other. The cytotoxic effects of OPE on HT-29 cells were confirmed by MTT and LDH release assays. LPS-induced oxidative conditions effectively downregulated TNF-α mRNA expression in OPE pretreated HT-29 cells compared with cells only stimulated with LPS. In addition, the expression of heme oxygenase-1 (HO-1) and glutathione S-transferase (GSTs) detoxification genes (i.e., GSTM1, GSTT1, and GSTP1) was upregulated after treatment with LPS at sublethal concentrations. However, the LPS-induced mRNA expression of HO-1 and GSTs was significantly attenuated by treatment with OPE. Therefore, onion peel extract is a promising component of future nutraceuticals and value-added products.     

山柰酚对人结肠癌SW48细胞增殖的抑制作用

目的为开发黄酮醇类抗肿瘤新药提供依据。方法采用噻唑蓝还原法检测山柰酚对SW48细胞生长的抑制作用,采用流式细胞仪分析山柰酚对细胞凋亡的诱导作用,采用免疫印记法分析山柰酚对凋亡通路相关蛋白表达的影响。结果山柰酚抑制SW48细胞的生长,上调了p53的蛋白表达量和磷酸化程度,改变了Bcl-2与Bax的蛋白含量比例。结论山柰酚通过诱导细胞周期阻滞和p53介导的线粒体凋亡通路抑制人结肠癌SW48细胞增殖。