中药逆转肿瘤细胞KBV200多药耐药活性体外筛选

选择100种常用于治疗癌症的中药，用MTT法体外筛选逆转肿瘤细胞KBV200多药耐药活性。其中葫芦、甘草、茯苓、穿心莲、山豆根、功劳木、艾叶、莪术、蛇床子等9种中药的乙醇提取物具有逆转活性，另有5种中药的提取物具有细胞毒作用。     

鬼臼毒素衍生物CIP-36诱导KBV200细胞凋亡

目的：研究鬼臼毒素衍生物CIP-36对多药耐药人口腔鳞状上皮癌细胞KBV200的抗肿瘤活性及其作用机制。方法：MTT法考察CIP-36对KBV200体外增殖的抑制作用;Giemsa染色、DNA ladder和流式细胞仪等方法进行细胞凋亡检测;免疫荧光法观察CIP-36对细胞骨架的作用;western-blot法检测CIP-36对KBV200细胞P-gp表达的影响。结果：CIP-36对KBV200细胞有明显的抑制作用,IC50值为（2.06±0.38）μmol/L,能够诱导细胞产生凋亡小体和DNAladder。流式细胞检测到了细胞凋亡峰,并观察到细胞周期出现S/G2＋M期阻滞。Western-blot结果显示P-gp表达降低,并且观察到CIP-36可破坏KBV200细胞的细胞骨架。结论：CIP-36可能通过降低P-gp的表达,破坏细胞骨架等多靶点克服KBV200细胞株的多药耐药性。     

鬼臼毒素衍生物CIP-36诱导KBV200细胞凋亡

目的研究鬼臼毒素衍生物CIP-36对多药耐药人口腔鳞状上皮癌细胞KBV200的抗肿瘤活性及其作用机制。方法MTT法考察CIP-36对KBV200体外增殖的抑制作用;Giemsa染色、DNAladder和流式细胞仪等方法进行细胞凋亡检测;免疫荧光法观察CIP-36对细胞骨架的作用;West-ernblot法检测CIP-36对KBV200细胞P-glycoprotein表达的影响。结果CIP-36对KBV200细胞有明显的抑制作用,IC50值为(2.06±0.38)μmol.L-1,能够诱导细胞产生凋亡小体和DNAladder。流式细胞检测到了细胞凋亡峰,并观察到细胞周期出现S/G2+M期阻滞。Westernblot结果显示P-gp表达降低,并且观察到CIP-36可破坏KBV200细胞的细胞骨架。结论CIP-36可能通过降低P-gp的表达,破坏细胞骨架等多靶点克服KBV200细胞株的多药耐药性。     

鬼臼毒素衍生物CIP-36抗肿瘤多药耐药的机制

研究鬼臼毒素衍生物CIP-36对耐药人口腔鳞状上皮癌细胞KBV200的抗肿瘤活性及其作用机制。采用MTT法观察CIP-36对多种肿瘤细胞和正常细胞的体外抑制作用以及对KB和KBV200细胞生长曲线的作用,Hoechst荧光染色进行细胞凋亡检测,RT-PCR检测CIP-36对KB及KBV200细胞p53、p21、caspase-3、bax、mdr-1和bcl-2的mRNA表达的影响,免疫组织化学检测观察CIP-36对KBV200细胞P-gp表达的影响。结果表明,CIP-36对多种肿瘤细胞均有较好的抑制作用,且对耐药株细胞均有明显抑制作用。荧光染色结果显示,CIP-36可诱导KBV200细胞凋亡。同时CIP-36可剂量依赖性地增加KBV200及KB细胞的p53、p21、caspase-3及bax的mRNA表达,同时降低mdr-1和bcl-2的mRNA表达,与对照组比较差异均有统计学意义。免疫组织化学检测结果显示,CIP-36显著降低KBV200的P-gp表达。提示CIP-36可能通过影响多个与肿瘤耐药相关基因和蛋白的表达来克服肿瘤细胞株的多药耐药性。     

中药秀雅杜鹃化学成分研究

摘要：目的:研究秀雅杜鹃的化学成分,评价分离化合物的药理作用。方法:对95%乙醇提取物采用乙酸乙酯萃取,对乙酸乙酯萃取部位采用硅胶柱、ODS柱色谱以及制备高效液相色谱进行分离纯化,并根据理化性质和核磁数据对化合物结构进行鉴定。采用MTT法检测化合物7对人喉癌上皮细胞Hep-2的药理活性。结果:共分离得到7个化合物,均为首次从该种属中分离得到,其中化合物2属于新天然产物。化合物7在高给药浓度(≥20μmol·L-1)时可以明显减弱人喉癌上皮细胞Hep-2细胞活性。结论:分离并确证了秀雅杜鹃乙酸乙酯部位化学成分,对化合物7的药理活性进行考察,为进一步研究秀雅杜鹃药用价值提供参考。     

鬼臼毒素衍生物CIP-36诱导KBV 200细胞凋亡

目的：研究鬼臼毒素衍生物CIP-36对多药耐药人口腔鳞状上皮癌细胞KBV 200的抗肿瘤活性及其作用机制。方法：MTT法考察CIP-36对KBV 200体外增殖的抑制作用;Giemsa染色、DNA ladder和流式细胞仪等方法进行细胞凋亡检测;免疫荧光法观察CIP-36对细胞骨架的作用;western-blot法检测CIP-36对KBV 200细胞P-gp表达的影响。结果：CIP-36对KBV 200细胞有明显的抑制作用,IC₅₀值为（2.06±0.38）μmol / L,能够诱导细胞产生凋亡小体和DNA ladder。流式细胞检测到了细胞凋亡峰,并观察到细胞周期出现S/G₂+M期阻滞。Western-blot结果显示P-gp表达降低,并且观察到CIP-36可破坏KBV 200细胞的细胞骨架。结论：CIP-36可能通过降低P-gp的表达,破坏细胞骨架等多靶点克服KBV 200细胞株的多药耐药性。     

山栀茶中一个新的三萜类化合物

为了研究山栀茶(Radix Pittospori)的化学成分,运用多种色谱方法进行分离纯化,从其氯仿提取物中分离得到2个三萜类化合物,并根据理化性质和波谱数据鉴定其结构分别为22-acetyl-21-(2-acetoxy-2-methylbutanoyl)-R1-barrigenol(1)和3α-hydroxyl-20-demethylisoaleuritolic-14(15)-ene-28,30-dioic acid(2)。其中,化合物1为新的三萜类化合物,化合物2为首次从该植物中分离得到。     

大黄中一个新的蒽醌苷(英文)

为了对药用大黄的根进行化学成分研究,采用硅胶、凝胶、反相等柱色谱和高效液相色谱方法分离和纯化化合物,根据理化性质和波谱方法鉴定化合物结构。从大黄的乙醇提取物中分离得到了1个新的和1个已知的蒽醌苷类化合物。此外,还测试了化合物对肝癌细胞的细胞毒活性。     

地榆中三萜类成分的研究

目的 研究地榆中的三萜类成分。方法 利用聚酰胺、硅胶、Sephadex LH-20和ODS柱色谱分离纯化,通过理化常数和波谱分析（¹H-NMR和¹³C-NMR）确定结构。结果 从体积分数70%乙醇提取物的乙酸乙酯萃取部分中分离鉴定了8个三萜类成分,分别为arjunic acid (1)、rosamultic acid (2)、haptadienic acid (3)、1β-羟基蔷薇酸 (4)、蔷薇酸 (5) 、 tormentic acid (6) 、坡模酸 (7) 和乌苏酸 (8)。结论 化合物1、2、3为首次从地榆属植物中分离得到的已知化合物。     

喙荚云实种子中的新二萜

目的研究喙荚云实的化学成分。方法采用多种色谱方法分离纯化,依据理化性质、波谱数据分析进行结构鉴定。结果从喙荚云实的体积分数为95%的乙醇回流提取物中分离鉴定了1个二萜类化合物。结论此二萜类化合物为新化合物。     

Reversal of multidrug resistance of KBV200 cells by triterpenoids isolated from Poria cocos

Multidrug resistance (MDR) of tumor cells is one of the major problems encountered during cancer chemotherapy. In this paper, we isolated eight triterpenoids from Poria cocos and evaluated their effects on reversing MDR of KBV200 cells. Eight triterpenoids increase significantly vincristine-induced cytotoxicity in drug-resistant KBV200 cells at the concentrations of 12.5 μg/mL and 25 μg/mL. Dehydrotumulosic acid showed the best reversal effect: it increased KBV200 apoptosis induced by vincristine and inhibited P-gp function through enhancing the accumulation and retention of fluorescent P-gp substrate rhodamine 123 in KBV200 cells but had no effect on P-gp expression.     

The multidrug resistant modulator HZ08 reverses multidrug resistance via P-glycoprotein inhibition and apoptosis sensitization in human epidermoid carcinoma cell line KBV200

Previous studies have demonstrated that the multidrug resistance modulator HZ08 has a strong multidrug resistance reversal effect in vitro and in vivo by inhibiting P-glycoprotein and multidrug resistance-associated protein 1 in K562/A02 and MCF-7/ADM cells, respectively. However, there are many other mechanisms responsible for resistance. In this study, MTT assay was used to examine the cytotoxicity and multidrug resistance reversal of HZ08 in KBV200 cells. It was also used to detect Rh123 and adriamycin accumulation in the presence of HZ08 to assess the effect on P-glycoprotein. Caspase-3 activity was analyzed under the incubation of HZ08 per se and in combination with vincristine. Results showed that HZ08 could increase the activity of caspase-3 with P-glycoprotein inhibition. Further studies revealed that HZ08 increased vincristine-induced apoptosis, characterized as an intrinsic apoptosis pathway with enhanced G2/M phase arrest, since HZ08 had an effect on the intrinsic apoptotic regulator Bcl-2 and Bax. Therefore, the outstanding reversal effect of HZ08 occurs not only through suppressing the P-glycoprotein function but also through activating the intrinsic apoptosis pathway.     

赛赓啶对 KBV200细胞多药抗性的逆转作用

研究赛赓啶对KB_V200细胞多药抗性的逆转作用及逆转机制。在KB_V200细胞,采用MTT法,测出赛赓啶对长春新碱、阿霉素和鬼臼乙叉甙耐药的逆转系数分别为5.5,2.0和1.9,而对5-氟尿嘧啶、美法仑的细胞毒性作用无明显影响,表明赛赓啶为多药抗性逆转剂。荧光分光光度法测定表明,赛赓啶可使KB_V200细胞内阿霉素蓄积量增加。流式细胞荧光测定显示赛赓啶可增加罗丹明123的蓄积并减慢其外排。免疫细胞化学及狭缝杂交表明赛赓啶不影响KB_V200细胞的P-糖蛋白染色深度和 mdr1 RNA 表达水平。以上结果提示赛赓啶的多药抗性逆转机制是抑制P-糖蛋白泵的功能。     

Two new lanostane triterpenoids from Poria cocos

Two new lanostane triterpenoids, 29-hydroxypolyporenic acid C (4) and 25-hydroxypachymic acid (5), together with three known compounds, ergosta-7,22-dien-3beta-ol (1), polyporenic acid C (2) and pachymic acid (3), were isolated from the 95% ethanolic extract of the sclerotium of Poria cocos (Schw.) Wolf. Their structures were determined by extensive spectroscopic analyses, including IR, UV, ESITOF-MS, HRESI-MS, 1D and 2D NMR data (1H NMR, 13C NMR, 1H-1H COSY, NOESY, HSQC and HMBC).     

KB细胞耐药株的建立及其耐药机制的探讨

用对长春新碱(VCR)敏感的KB细胞为亲本,通过诱变剂甲基磺酸乙酯刺激,然后在培养液中加入浓度递增的CVR,得到耐药细胞株KB_V200。此细胞株对VCR的耐受程度约为KB细胞的175倍。对其它抗肿瘤药物如紫杉醇、秋水仙碱和阿霉素等也有不同程度的交叉耐药性。进一步研究表明,KB_V200对³H-VCR的蓄积明显减少,且耐药基因(mdr1)表达增加。钙通道阻滞剂维拉帕米(Ver)可增加KB_V200对³H-VCR的蓄积和对VCR的敏感性。这些结果提示,KB_V200耐药的机制可能是由于mdr1基因表达增加,产生过量的p-糖蛋白,使药物外排增多所致。     

Marine sponge-derived sipholane triterpenoids reverse P-glycoprotein (ABCB1)-mediated multidrug resistance in cancer cells

Previously, we reported sipholenol A, a sipholane triterpenoid from the Red Sea sponge Callyspongia siphonella, as a potent reversal of multidrug resistance (MDR) in cancer cells that overexpressed P-glycoprotein (P-gp). Through extensive screening of several related sipholane triterpenoids that have been isolated from the same sponge, we identified sipholenone E, sipholenol L and siphonellinol D as potent reversals of MDR in cancer cells. These compounds enhanced the cytotoxicity of several P-gp substrate anticancer drugs, including colchicine, vinblastine and paclitaxel, and significantly reversed the MDR-phenotype in P-gp-overexpressing MDR cancer cells KB-C2 in a dose-dependent manner. Moreover, these three sipholanes had no effect on the response to cytotoxic agents in cells lacking P-gp expression or expressing MRP1 (ABCC1) or MRP7 (ABCC10) or breast cancer resistance protein (BCRP/ABCG2). All three sipholanes (IC₅₀ >50 μM) were not toxic to all the cell lines that were used. [³H]-Paclitaxel accumulation and efflux studies demonstrated that all three triterpenoids time-dependently increased the intracellular accumulation of [³H]-paclitaxel by directly inhibiting P-gp-mediated drug efflux. Sipholanes also inhibited calcein-AM transport from P-gp-overexpressing cells. The Western blot analysis revealed that these three triterpenoids did not alter the expression of P-gp. However, they stimulated P-gp ATPase activity in a concentration-dependent manner and inhibited the photolabeling of this transporter with its transport substrate [¹²⁵I]-iodoarylazidoprazosin. In silico molecular docking aided the virtual identification of ligand binding sites of these compounds. In conclusion, sipholane triterpenoids efficiently inhibit the function of P-gp through direct interactions and may represent potential reversal agents for the treatment of MDR.