雷公藤内酯醇对IgA肾病大鼠蛋白尿和nephrin、podocin蛋白及mRNA表达的影响

目的:观察雷公藤内酯醇(TP)对IgA肾病(IgAN)大鼠尿蛋白和足细胞nephrin、podocin蛋白及mRNA表达的影响。方法:采用牛血清白蛋白+四氯化碳+脂多糖的方法建立IgAN大鼠模型。将60只大鼠随机分为正常组、模型组、洛丁新组、TP剂量组、TP中剂量组、TP高剂量组,每组10只。于0,10,14周收集血尿标本,并处死大鼠留取肾组织标本。检测24 h尿蛋白定量,血生化指标,分别采用实时定量PCR及ELISA法检测肾组织nephrin、podocin mRNA和蛋白表达。结果:(1)实验前各组大鼠尿蛋白差异无统计学意义(P>0.05),第10周末各造模组蛋白尿均明显高于正常组(P<0.01),第14周末TP各剂量组及洛丁新组蛋白尿均较模型组明显减少(P<0.01),TP高剂量组Alt值高于其余各组(P<0.05);(2)与正常组相比,模型组nephrin、podocin蛋白、mRNA表达明显降低(P<0.01);TP各剂量组及洛丁新组nephrin、podocin蛋白、mRNA表达均较模型组明显增高(P<0.05);TP中、高剂量组较洛丁新组nephrin、podocin蛋白浓度显著增高(P<0.05);TP各剂量组与洛丁新组及TP各剂量组间nephrin、podocin mRNA表达均差异无统计学意义(P>0.05)。结论:TP能够明显减轻IgAN大鼠的蛋白尿,且能明显提高肾组织nephrin、podocin mRNA及蛋白的表达。提示TP能调节IgAN足细胞相关分子nephrin、podocin基因及蛋白的表达,减少足细胞损害,减少尿蛋白。     

雷公藤内酯醇诱导胰腺癌细胞凋亡及对5-脂氧合酶和凋亡基因表达的影响

目的探讨雷公藤内酯醇(TL)在胰腺癌化学预防和治疗中的应用价值。方法采用人胰腺癌细胞株SW1990作为研究对象,MTT法和吖啶橙(AO)染色分别检测SW1990细胞的增殖和凋亡,采用半定量RT-PCR法检测5-脂氧合酶(5-LOX)mRNA、bcl-2 mRNA和bax mRNA的表达。结果10μg/ml的TL对SW1990细胞生长的抑制率达30%以上,0.1μg/mlTL处理24h后,SW1990细胞凋亡指数达38.5%,显著高于对照组的14.97%。5-LOX、bcl-2、bax mRNA表达从0.7160±0.0251,0.2446±0.0311和0.0260±0.0065分别变化为0.2400±0.0316,0.0700±0.0158和0.0700±0.0158。结论TL能抑制胰腺癌细胞增殖和诱导细胞凋亡,其凋亡机制可能与下调5-LOXmRNA和bcl-2mRNA、上调bax mRNA基因表达有关。     

雷公藤内酯醇对神经胶质瘤U87细胞侵袭性抑制的体外研究

目的 研究雷公藤内酯醇（triptolide）对体外培养的胶质瘤U87细胞侵袭性的抑制作用,并探讨其作用分子机制. 方法 使用MTT法检测雷公藤内酯醇对胶质瘤U87细胞增殖抑制作用, Transwell实验检测雷公藤内酯醇处理后细胞侵袭能力的变化,进一步通过Real-Time PCR和Western印迹法检测雷公藤内酯醇对U87胶质瘤细胞的组织蛋白酶B（Cathepsins B,CB）、基质金属蛋白酶9（MMP-9）表达的影响. 结果 雷公藤内酯醇以剂量-效应方式抑制胶质瘤细胞增殖,同时可减弱U87胶质瘤细胞的侵袭能力,下调其CB、MMP-9表达水平. 结论 在体外实验中雷公藤内酯醇抑制U87胶质瘤细胞侵袭性生长的机制与下调CB及MMP-9的表达,降低胶质瘤细胞水解细胞外基质的能力有关.     

Apoptosis of human pancreatic cancer cells induced by Triptolide

AIM： To investigate apoptosis in human pancreatic cancer cells induced by Triptolide （TL）, and the relationship between this apoptosis and expression of caspase-3＇ bcl-2 and bax. METHODS： Human pancreatic cancer cell line SW1990 was cultured in DMEM media for this study. MTT assay was used to determine the cell growth inhibitory rate in vitro. Flow cytometry and TUNEL assay were used to detect the apoptosis of human pancreatic cancer cells before and after TL treatment. RT-PCR was used to detect the expression of apoptosis-associated gene caspase-3＇ bcl-2 and bax. RESULTS： TL inhibited the growth of human pancreatic cancer cells in a dose-and time-dependent manner. TL induced human pancreatic cancer cells to undergo apoptosis with typically apoptotic characteristics. TUNEL assay showed that after the treatment of human pancreatic cancer cells with 40 ng/mL TL for 12 h and 24 h, the apoptotic rates of human pancreatic cancer cells increased significantly. RT-PCR demonstrated that caspase-3 and bax were significantly up-regulated in SW1990 cells treated with TL while bcl-2 mRNA was not. CONCLUSION： TL is able to induce the apoptosis in human pancreatic cancer cells. This apoptosis may be mediated by up-regulating the expression of apoptosisassociated caspase-3 and bax gene.     

雷公藤甲素对哮喘大鼠气道平滑肌增生及c-fos与c-jun表达的影响

目的：探讨雷公藤甲素（TL）对哮喘大鼠气道平滑肌（ASM）增生及原癌基因c-fos与c-jun基因表达的影响。方法：选择70只雄性SD大鼠随机分为7组，每组10只，采用卵蛋白腹腔注射及雾化吸入制成大鼠哮喘模型，采用志录－聚合酶链反应（RT－PCR）法检测各组的c-fos与c-jun的mRNA表达水平，同时用免疫组化方法检测c-fos与c-jun蛋白表达水平，并在光镜下观察支气管壁厚度（WA／Pi)，支气管平滑肌厚度（平滑肌面积／Pi)，支气管平滑肌细胞核数量（N／Pi)及肺组织形态学改变，结果：RT－PCR和免疫组化均显著哮喘组（A组），治疗组（C，D，E，F，G组）c-fos与c-jun的mRNA和蛋白表达与对照组（B组）比较，差异有显著性（P＜0．01）；各治疗组的表达与A组比较差异也有显著性（P＜0．01），而各治疗组之间比较差异无显著性（P＞0．05），是结果显示，TL减轻ASM的增生。结论：ASM增生肥厚是哮喘的一个显著特征，c-fos及c-jun可能在此起着促进作用，TL通过下调c-fos和c-jun表达而起抑制ASM增生作用。     

雷公藤内酯醇对原发免疫性血小板减少症患者浆细胞样树突细胞功能及成熟的影响

目的 探讨雷公藤内酯醇对原发免疫性血小板减少症(ITP)患者浆细胞样树突细胞(pDC)功能及成熟的影响.方法 取22例初治ITP患者(初治组)、22例精皮质激素治疗后完全缓解(CR)ITP患者(CR组)及22名健康志愿者(正常对照组)外周静脉血,分离单个核细胞,用流式细胞术分选pDC,分别加入0、5、10、30μg/L的雷公藤内酯醇共孵育.24 h后收集上清液,用ELISA法检测IFN-α、IL-6、TNF-α水平;5d后收集细胞,用流式细胞术检测髓样树突细胞(mDC)CD11c、CD80、CD86表达,光镜下观察mDC的形态,电镜观察mDC的超微结构.结果 加入10μg/L雷公藤内酯醇后初治组IFN-α、IL-6、TNF-α水平分别为(451.32±85.77)、(105.68±23.85)、(135.78±30.62)ng/L,CR组分别为(391.71±72.49)、(84.73±17.77)、(108.16±23.21)ng/L,正常对照组分别为(335.51±67.54)、(73.62±21.82)、(95.58±32.85)ng/L,初治组及CR组高于正常对照组(P<0.05),初治组高于CR组(P<0.05),并呈雷公藤内酯醇浓度依赖性(P<0.05);初治组及CR组mDC表型CD11c、CD80、CD86阳性率高于正常对照组(P<0.05),初治组高于CR组(P<0.05),加入雷公藤内酯醇后mDC表型阳性率呈浓度依赖性降低(P<0.05).加入雷公藤内酯醇后mDC较对照组突起变少、变短,形态上不成熟.结论 雷公藤内酯醇能够减弱ITP患者pDC的功能,并抑制其向mDC的分化和成熟.     

雷公藤内酯醇抗人肝癌SMMC-7721细胞活性研究

目的: 评价雷公藤内酯醇抗人肝癌SMMC-7721细胞的作用.方法: 采用体外细胞培养和SD大鼠体内肿瘤接种两种衡量方法,体外细胞培养检测指标包括细胞形态观察,台盼蓝染色,DAPI染色,DNA ladder;SD大鼠体内接种肿瘤指标检测用游标卡尺测量肿瘤体积大小.结果: 与对照组相比,体外培养肿瘤细胞在用药后活细胞大幅减少,体内接种肿瘤细胞在用药后,肿瘤块体积较小且增长缓慢,而对照组肿瘤体积却大幅增长.结论: 雷公藤内酯醇对体外和体内培养的人肝癌SMMC-7721细胞有明显的生长抑制作用和促凋亡作用.     

Triptolide and l-ascorbate palmitate co-loaded micelles for combination therapy of rheumatoid arthritis and side effect attenuation

Triptolide (TP) has its unique curative effect in the treatment of rheumatoid arthritis (RA), but its application is limited by the poor water solubility and multi-organ toxicity. We herein developed a novel nanoparticle platform composed of L-ascorbate palmitate (VP, vitamin C derivative) that can deliver TP to synergistically treat arthritis and inhibit the occurrence of oxidative stress. The TP-loaded nanoparticles (termed TP-VP NPs) showed the suitable particle size (about 145 nm) and good physical stability. TP-VP NPs effectively down-regulated IL-1β, IL-6 and TNF-α levels to inhibit the erosion of synovitis and bone tissue, and alleviate the swelling and deformation of CIA mice’s feet. Compared to the TP, TP-VP NPs could inhibit effectively the oxidative stress in liver, and alleviate significantly the triptolide-induced toxicity injury in liver, kidney and testicle. The results demonstrated that TP-VP NPs is a promising triptolide delivery system for the treatment of RA, which enhances the water solubility of TP and reduces the toxicity of TP in vivo.     

雷公藤内酯醇与人乳腺癌细胞大分子DNA作用模式的探讨

采用药物与DNA作用的体外检测方法，以DNA嵌入剂表阿霉素（EDR）作对照，从分子药理水平研究雷公藤内酯醇（TL）对乳腺癌细胞DNA的作用模式。实验发现TL在120μg／ml浓度，37℃作用24h可与DNA模板结合，同EDR呈嵌合效应，但嵌入DNA尚不牢固，易发生解离；EDR在较低浓度（50μg／ml）作用30min即可显示明显的嵌合效应且是不可逆的。说明TL对乳腺癌细胞DNA的作用模式为嵌合逆行模型，嵌合能力远低于EDR，与DNA结合亦不可能是其抗癌主效应。     

Mechanism of triptolide-induced apoptosis: effect on caspase activation and Bid cleavage and essentiality of the hydroxyl group of triptolide

Triptolide is a compound extracted from the Chinese herb Tripterygium wilfordii Hook. f. Triptolide has potent anticancer activity. However, the mechanisms by which triptolide exerts its anticancer activities remain unclear. To explore the molecular mechanisms involved in the anticancer activity of triptolide, we have examined the effect of triptolide on the growth of pancreatic carcinoma PANC-1 and cervical adenocarcinoma HeLa cells. We found that treatment of both HeLa and PANC-1 cells with triptolide potently suppressed cell growth and induced apoptosis, indicated by nuclear fragmentation and blebbing. In both HeLa and PANC-1 cells, apoptosis induced by triptolide was associated with activation of both caspase-3 and caspase-8, and cleavage of poly(ADP-ribose) polymerase and Bid. Moreover, in HeLa cells, caspase-9 is also significantly activated in response to triptolide. Overexpression of Bcl-2 in HeLa cells substantially attenuated triptolide-induced apoptosis. Interestingly, substitution of the 14-OH of triptolide with an acetyl group abrogated both its anticancer and its antiinflammatory activities. Our studies suggest that triptolide may exert its anticancer effects by initiating apoptosis through both death-receptor- and mitochondria-mediated pathways. Our results indicate that both the apoptosis-promoting and the antiinflammatory activities of triptolide depend on the 14-OH group.