Triptolide and management of systemic malignancies besides pancreatic carcinomas

The recent article by Zhou et al was highly interesting and thought provoking. The authors have clearly shown that triptolide administration is associated with upregulation of the Bax gene, resulting in an attenuating effect on cell growth in gastrointestinal malignancies such as pancreatic carcinomas. The article by Zhou etal is all the more important because it highlights the rapidly increasing role of triplodide in the management of systemic malignancies. For instance, triptolide acts on the PI3K/Akt/NF-κB pathway, thereby enhancing apoptosis secondary to the administration of bortezomib in multiple myeloma cells. Similar synergisms are seen when triptolide is administered along with 5-fluoruracil for the management of colonic carcinomas. Similarly, triptolide causes down-regulation of the Bcl-2 gene, resulting in control of cell growth in tumors, such as glioblastoma multiformes.     

姜黄素雷公藤内酯醇对前列腺癌细胞PC3及血管内皮生长因子影响的比较

目的研究姜黄素（curcumin,Cur）、雷公藤内酯醇（triptolide,TL）对非雄激素依赖性前列腺癌细胞株PC3细胞体外作用及其血管内皮生长因子（vascular endothelial growth factor,VEGF）表达的影响。方法分别用梯度浓度的Cur和TL作用于PC3细胞,MTT法检测细胞生长活性;流式细胞仪测定细胞周期及凋亡的变化;半定量RT-PCR法检测PC3细胞内VEGF mR-NA的表达;ELISA检测细胞上清液中分泌VEGF蛋白的浓度。结果Cur及TL都能呈剂量与时间依赖性显著抑制PC3细胞的生长,不同浓度组之间及不同作用时间组之间的差异均有统计学意义（P〈0.01）。Cur、TL诱导PC3细胞分别出现剂量依赖性G2/M、S期阻滞（P〈0.01）,且各浓度组凋亡细胞比例差异有统计学意义（P〈0.01）;PC3细胞内VEGF mRNA的表达和细胞上清液中分泌VEGF蛋白亦呈剂量依赖性降低。结论Cur、TL能显著抑制体外PC3细胞的生长,分别促进细胞周期阻滞于不同时期,增加凋亡,并且VEGF mRNA及蛋白的表达明显降低,两药抑制肿瘤和血管生长机制不同。     

基于狗肾小管上皮细胞的雷公藤甲素体外肾毒性研究

目的:观察雷公藤甲素对狗肾小管上皮细胞(MDCK细胞)的毒性作用,并初步探讨其对氧化应激的影响。方法:以马兜铃酸A为阳性对照,用0.5,5,50和500 nmol·L-1的雷公藤甲素与MDCK细胞共同孵育24 h后,MTT法检测细胞抑制率,乳酸脱氢酶(LDH)释放实验检测细胞膜损伤以及倒置显微镜观察雷公藤甲素对细胞形态的改变。用500 nmol·L-1雷公藤甲素分别与MDCK细胞作用30 min、1 h、2 h、4 h和6 h后,用2’,7’-二氯荧光黄双乙酸盐(DCFH-DA)荧光探针检测细胞的活性氧自由基(ROS)水平。结果:与阴性对照组比较,雷公藤甲素组的细胞抑制率明显升高(P<0.01);LDH相对释放率明显增加(P<0.01)。雷公藤甲素处理组的细胞形态出现皱缩,呈现为球形,并有部分细胞脱落。雷公藤甲素与MDCK细胞作用30 min后ROS水平达到最高值,然后ROS逐渐减少(P<0.01)。结论:雷公藤甲素可诱导MDCK细胞的毒性作用,其毒性作用机制可能和氧化应激反应有关。     

The Roles of Plant-Derived Triptolide on Non-Small Cell Lung Cancer

Over the past decade, natural compounds have been proven to be effective against many human diseases, including cancers. Triptolide (TPL), a diterpenoid triepoxide from the Chinese herb Tripterygium wilfordii Hook F, has exhibited attractive cytotoxic activity on several cancer cells. An increasing number of studies have emphasized the antitumor effects of TPL on non-small cell lung cancer (NSCLC). Here we mainly focused on the key molecular signaling pathways that lead to the inhibitory effects of TPL on human NSCLC, such as mitogen-activated protein kinases (MAPKs) modulation, inhibition of NF- B activation, suppression of miRNA expression, etc. In addition, the effect of TIG on immune response in cancer patients is summarized for improved immune modulation utilization. However, the clinical use of TPL is often limited by its severe toxicity and water insolubility. Future clinical trials and drug delivery strategies that will evaluate the security and validate the underlying tumor-killing properties of TPL in human NSCLC are also to be discussed.     

Triptolide inhibits murine-inducible nitric oxide synthase expression by down-regulating lipopolysaccharide-induced activity of nuclear factor-kappa B and c-Jun NH2-terminal kinase

Triptolide (PG490) is a natural, biologically active compound extracted from the Chinese herb Tripterygium wilfordii. It has been shown to possess potent anti-inflammatory and immunosuppressive properties. In Raw 264.7 cells stimulated with lipopolysaccharide (LPS) to mimic inflammation, triptolide inhibits nitric oxide (NO) production in a dose-dependent manner and abrogates inducible nitric oxide synthase (iNOS) gene expression. To investigate the mechanism by which triptolide inhibits murine iNOS gene expression, we examined activation of mitogen-activated protein kinases (MAP kinases) and nuclear factor-kappa B (NF-kappa B) in these cells. Addition of triptolide inhibited phosphorylation of c-Jun NH(2)-terminal kinase (JNK) but not that of extracellular signal-regulated kinase (ERK) or p38 mitogen-activated protein kinase. In addition, triptolide significantly inhibited the DNA binding activity of NF-kappa B. Taken together, these results suggest that triptolide acts to inhibit inflammation through inhibition of NO production and iNOS expression through blockade of NF-kappa B and JNK activation.     

昆明山海棠雄性抗生育成分的研究

昆明山海棠茎皮乙醇提取物中雄性抗生育活性成分的分离，显示氯仿部分及正丁醇部分为活性部分。用硅胶柱对这两部分进一步分离后，得到雷藤素乙（Ⅰ）、雷藤素甲（Ⅱ）、雷酚二萜酸（Ⅲ）、雷酚萜醇（Ⅳ）、3－oxo-olean-12-en-29-oicacid(Ⅴ）、齐墩果酸（Ⅵ）、3β，22α-dihydroxy-△^12-oleanen-29-oic acid(Ⅶ）、3－乙酰氧基齐墩果酸（Ⅷ）、雷公藤内酯甲（Ⅸ）、雷公藤次硷（Ⅹ）、胡萝卜甙（Ⅺ）、β－谷甾醇（Ⅻ）、1－表儿茶素（ⅩⅢ）及富马酸（ⅩⅣ）。其中3－oxo-olean-12-en-29-oic acid，齐墩果酸及富马酸为首次从雷公藤属植物中发现。雷藤素乙。雷酚萜醇，3－β，22α-dihydroxy-△^12-oleanen-29-oic acid及胡萝卜甙为首次从昆明山海棠中分离得到。药理筛选显示雷藤素乙及雷藤素甲对于雄性大鼠具有可逆性雄性抗生育活性，其有效剂量分别为每天200及30μg/kg。     

Inhibitory effects of Triptolide on interferon-gamma-induced human leucocyte antigen-DR, intercellular adhesion molecule-1, CD40 expression on retro-ocular fibroblasts derived from patients with Graves' ophthalmopathy

PURPOSE: To explore the effects of Triptolide, the principal active diterpenoid from the Chinese Medicinal Herb Tripterygium Wilfordii Hook F that has immunosuppressive and anti-inflammatory properties, on cell proliferation, hyaluronic acid (HA) synthesis, and the expressions of human leucocyte antigen-DR (HLA-DR), intercellular adhesion molecule-1 (ICAM-1) and CD40 on cultured retro-ocular fibroblasts (RFs) from patients with Graves' ophthalmopathy. METHODS: After two to five passages, cultured RFs were incubated for 48 h within a medium alone or in the presence of recombinant human interferon-gamma (IFN-gamma) and various concentrations of Triptolide. Cell viability was assessed by MTT (3-[4.5-dimethylahiazol-2-yl]-2,5-diphenyltetrazolium Bromide). RFs proliferation was assessed by [(3)H]-thymidine incorporation assay. Flow cytometry was used to investigate the amount of HLA-DR, ICAM-1 and CD40. HA synthesis was measured by radioimmunoassay. RESULTS: Cell viability was not detrimentally affected when incubated with Triptolide from 0.01 microg/L to 10 microg/L for 48 h, and decreased with 20 microg/L Triptolide. The incorporation of [(3)H]-thymidine of RFs was 55 476 +/- 15 842 cpm incubated with medium alone or 18 352 +/- 3568 cpm with 10 microg/L Triptolide (t = 5.600, P < 0.01). Initially, the percentage of positive cells of HLA-DR, ICAM-1 and CD40 on RFs were 4.75 +/- 2.13%, 17.53 +/- 10.12% and 6.38 +/- 2.23%, respectively, and the synthesis of HA was 100 +/- 12%. Compared with basal values, 48-h incubation with IFN-gamma (100 U/mL) significantly enhanced the amount of HLA-DR, ICAM-1 and CD40, and HA synthesis. The values were 60.58 +/- 10.12% (t = 13.224, P < 0.01), 62.66 +/- 18.17% (t = 5.315, P < 0.01), 57.67 +/- 13.61% (t = 9.110, P < 0.01) and 164 +/- 22% (t = 9.238, P < 0.01), respectively. Triptolide 0.01 microg/L had little effect on IFN-gamma-induced HLA-DR, ICAM-1 and CD40 amounts, as well as HA synthesis. When the concentration ranged from 0.1 microg/L to 10 microg/L, Triptolide inhibited IFN-gamma-induced RFs activation in a dose-dependent manner. It was also found that Triptolide had the same inhibiting effects on IFN-gamma-induced RFs and skin fibroblasts from patients with normal individual conditions. CONCLUSIONS: Triptolide could inhibit IFN-gamma-induced activation of RFs derived from patients with Graves' ophthalmopathy.     

雷公藤内酯醇对牛晶状体上皮细胞增殖细胞核抗原表达的影响

目的:观察雷公藤内酯醇(Triptolide,Tri)对体外培养的牛晶状体上皮细胞(bovine Lens epithelial cell,BLECs)增殖细胞核抗原的影响和对BLECs增殖的作用。方法:取第4代对数生长期的牛LECs,培养24h后,加入重组人表皮生长因子(Recombinant Human Epithelial Growth Factor,rhEGF)(终浓度50μg/L)和不同浓度的Tri(40,20,10μg/L),再分别作用6,12,24,48,72h后,采用流式细胞仪检测增殖细胞核抗原(PCNA)在LECs的阳性表达率及增殖抑制率。结果:不同浓度Tri(40～10μg/L)抑制处于增殖状态的LECs,PCNA表达率在6～72h内随浓度增高和作用时间延长有明显的下调,10μg/L组6hPCNA表达是(76.74±0.8),与同时间增殖对照组比较有显著性差异(P<0.05),12h(70.32±1.6),24h(61.96±2.1),48h(54.98±2.0),72h(42.15±1.6);20μg/L组6hPCNA表达是(68.58±3.2),12h(55.34±2.6),24h(49.59±1.6),48h(36.61±0.5),72h(30.00±1.1);40μg/L组6hPCNA表达是(60.96±1.2),12h(46.45±1.7),24h(32.10±0.5),48h(27.15±1.1),72h(19.16±1.5),以上各组与同时间增殖对照组比较均有非常显著性差异(P<0.01),呈明显的时间—效应关系和剂量—效应关系。结论:雷公藤内酯醇使牛晶状体上皮细胞内PCNA的表达率下降,改变细胞周期,从而抑制细胞增殖。     

雷公藤内酯醇抑制克罗恩病肠道浆膜纤维细胞黏附分子的表达

目的:观察雷公藤内酯醇对克罗恩病病人体外肠道浆膜纤维细胞黏附分子(ICAM-1)表达的影响. 方法:取4例克罗恩病病人手术中肠浆膜纤维细胞,体外培养3～5代.用不同浓度(0～60 ng/mL)的雷公藤内酯醇进行预处理1 h,然后加入IL-1β (10 ng/mL)共同孵育24 h.分别使用流式细胞仪和蛋白质印迹法测定细胞表面和细胞内ICAM-1的表达. 结果:IL-1β可诱导克罗恩病肠浆膜纤维细胞ICAM-1表达水平显著升高,而雷公藤内酯醇能显著抑制IL-1β的作用,并呈浓度依赖性. 结论:雷公藤内酯醇可抑制IL-1β诱导的克罗恩病肠浆膜纤维细胞ICAM-1的表达.     

雷公藤内酯醇对实验性变态反应性脑脊髓炎大鼠外周血SIL-2R表达的影响

目的:探讨雷公藤内酯醇对实验性变态反应性脑脊髓炎(EAE)大鼠外周血清SIL-2R表达的影响。方法:模型组采用豚鼠髓鞘蛋白匀浆和福氏完全佐剂诱发大鼠急性EAE,治疗组在模型组基础上予以不同剂量雷公藤内酯醇[0.2mg/(kg.d),0.4mg/(kg.d)],观察临床表现并评分;HE染色和Loyez氏髓鞘染色观察病理和髓鞘改变;ELISA法测定血清的SIL-2R水平。结果:高剂量组未出现临床症状;低剂量组EAE临床症状较模型组轻。高剂量组SIL-2R浓度为(36.43±5.43)mg/mL,与模型组和低剂量Tri组比较P<0.01和P<0.05;脑和脊髓小血管周围炎症细胞浸润明显减少;髓鞘结构完整。结论:雷公藤内酯醇对EAE的治疗有剂量相关性,其作用机制与抑制SIL-2R表达有关。