雷公藤红素对裸鼠肝癌皮下移植瘤抑制作用的研究

目的 研究天然活性化合物雷公藤红素（celastrol）对裸鼠移植肿瘤的抑制作用。方法 建立人源肝癌HepG2细胞的裸鼠皮下移植肿瘤模型。20只裸鼠随机分为4组,即空白对照组和雷公藤红素高剂量组、中剂量组和低剂量组,每组分别按4.0 mg/kg、2.0 mg/kg、1.0 mg/kg雷公藤红素腹腔给药,每日一次,每周5 d,共给药30 d,实验中每5 d测量一次肿瘤体积的变化。结果 雷公藤红素高剂量组、中剂量组和低剂量组的体积抑瘤率分别是89.52%、65.60%和12.21%,与空白对照组相比,高剂量组、中剂量组的差异具有统计学意义（P〈0.05）。高剂量雷公藤红素给药后对裸鼠肝脏和肾脏等产生严重的毒性反应。结论 雷公藤红素对裸鼠肝癌皮下移植瘤具有良好的体内抗肿瘤效果,应进一步开展药物新剂型及分子结构优化方面的研究,减少其毒副反应。     

雷公藤红素对C6胶质瘤细胞凋亡及细胞周期阻滞的影响

目的 探讨雷公藤红素对体外C6胶质瘤细胞凋亡及细胞周期阻滞的影响及机制.方法 雷公藤红素处理体外培养的C6胶质瘤细胞,MTT法检测细胞增殖,Hochest33342染色荧光显微镜观察以及锇铀铅染色透射电镜观察胶质瘤细胞形态变化,流式细胞术分析细胞凋亡率及细胞周期改变.蛋白印迹分析检测凋亡相关蛋白及细胞周期调控蛋白的表达变化.结果 雷公藤红素对C6胶质瘤细胞的增殖抑制作用呈浓度和时间依赖性;2.56 μmol/L雷公藤红素处理C6胶质瘤细胞24h后,荧光显微镜、透射电镜下均可见凋亡形态学改变.流式细胞术检查显示,凋亡细胞比例由1.32％升高到19.34％;并且,处于G0/G1期的细胞比例由76.42％降低到54.52％,G2/M期细胞比例由9.29％升高到30.50％.蛋白印迹分析显示,雷公藤红素降低了Bcl -2及XIAP蛋白表达,促进了Bax及Caspase -3的表达以及PARP的剪切;雷公藤红素在诱导细胞周期调控蛋白p21、p27及cyclin B1蛋白表达增加的同时抑制了cdk2蛋白的表达.结论 雷公藤红素能够通过调节凋亡相关蛋白及细胞周期相关因子的表达影响C6胶质瘤细胞的凋亡及细胞周期阻滞.     

The NF-κB inhibitor celastrol attenuates acute hepatic dysfunction induced by cecal ligation and puncture in rats

Acute hepatic dysfunction associating sepsis is mediated mainly by toll-like receptor-4 (TLR-4)/nuclear factor kappa‐B (NF-κB) inflammatory pathway. This study explores potential hepatoprotective effect of the NF-κB inhibitor celastrol in cecal ligation and puncture (CLP) model in rats.Protective effect of celastrol (1 mg/kg, i.p., 1 h before CLP) was illustrated after 24 h by preventing CLP‐induced hepatic histopathological changes and elevation in serum hepatic biomarkers [alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TB) and gamma aminotransferase (γ-GT)] without affecting mortality. Celastrol anti‐inflammatory effect was illustrated by inhibiting increased serum and hepatic mRNA expression of interleukin-6 (IL-6) without affecting IL-10 elevation. Furthermore, celastrol inhibited CLP-induced elevations in hepatic mRNA expression of nuclear factor inhibitory protein kappa-B alpha (NFκBia), TLR-4, 5‐lipoxygenase (5-LOX) and prevented NF‐κB/p65 nuclear translocation and activation.In conclusion, celastrol prevented CLP-induced acute hepatic dysfunction through its anti-inflammatory effect by attenuating NF-κB activation, TLR-4 and 5-LOX expression with subsequent reduction in pro-inflammatory IL-6.     

The MVA pathway genes expressions and accumulation of celastrol in Tripterygium wilfordii suspension cells in response to methyl jasmonate treatment

Celastrol is an important bioactive triterpenoid in traditional Chinese medicinal plant, Tripterygium wilfordii. Methyl Jasmonate (MJ) is a common plant hormone which can regulate the secondary metabolism in higher plants. In this study, the mevalonate (MVA) pathway genes in T. wilfordii were firstly cloned. The suspension cells of T. wilfordii were elicited by MJ, and the expressions of MVA pathway genes were all enhanced in different levels ranging from 2.13 to 22.33 times of that at 0 h. The expressions were also enhanced compared with the CK group separately. The accumulation of celastrol in the suspension cells after the treatment was quantified and co-analyzed with the genes expression levels. The production of celastrol was significantly increased to 0.742 mg g^?1 after MJ treatment in 288 h which is consistent with the genes expressions. The results provide plenty of gene information for the biosynthesis of terpenoids in T. wilfordii and a viable way to improve the accumulation of celastrol in T. wilfordii suspension cells.     

雷公藤红素对多发性骨髓瘤细胞株RPMI 8226的体外抑制作用研究

目的研究雷公藤红素（Celastrol）对多发性骨髓瘤细胞株RPMI 8226的抑制作用;方法使用CCK-8试剂研究生长抑制作用,使用An-nexin-V/PI双染色法,研究诱导凋亡作用;使用流式细胞仪进行sub-G1峰及细胞周期分析;结果雷公藤红素能够抑制RPMI 8226细胞生长,呈现剂量依赖性,IC50值为1.87μmol.L-1;通过Annexin-V/PI双染色法以及sub-G1峰分析,证实雷公藤红素可以诱导肿瘤细胞凋亡;并使肿瘤细胞的S期细胞减少;结论雷公藤红素可以抑制骨髓瘤细胞株RPMI 8226生长,诱导细胞凋亡,并使S期细胞减少。     

Celastrol,a novel HSP90 inhibitor,depletes Bcr–Abl and induces apoptosis in imatinib-resistant chronic myelogenous leukemia cells harboring T315I mutation

T315I Bcr–Abl in chronic myelogenous leukemia (CML) is the most notorious point mutations to elicit acquired resistance to imatinib. In the present study, we investigated the effect of celastrol on CML cells bearing wild-type Bcr–Abl or T315I-mutant. The results revealed that celastrol potently downregulated the protein levels of Bcr–Abl, and inhibited the growth in CML cells in vitro and in nude mouse xenografts regardless of Bcr–Abl mutation status. Celastrol induced mitochondrial-dependent apoptosis. In conclusion, celastrol exhibits potent activity against CML cells bearing wild-type Bcr–Abl or -the T315I-mutant.     

雷公藤红素:一种潜在的治疗肥胖症的新药物

雷公藤红素是存在于传统中药雷公藤根部的一种苯醌甲基三萜系化合物.近几年的研究发现,雷公藤红素能够减少摄食量、增加能量消耗,从而发挥强效的减重作用.其作用机制包括激活瘦素受体-信号转导与转录激活因子3信号通路和减轻下丘脑内质网应激,从而加强瘦素的作用;通过激活热休克因子1-过氧化物酶体增殖物活化受体γ协同刺激因子1α(HSF1-PGC1α)轴,促进能量代谢;通过抑制核因子-κB信号通路,改善胰岛素抵抗.雷公藤红素的上述作用使其有望成为未来治疗肥胖症的一种新药物.     

雷公藤红素对人前列腺癌细胞凋亡及SENP1基因表达的影响

目的：研究雷公藤红素对人前列腺癌（PCa）细胞生长、凋亡及SUMO特异蛋白酶1（SUMO-specificproteases 1,SENP1）基因表达的影响。方法：对前列腺癌细胞PC-3和LNCaP进行雷公藤红素处理,通过测定细胞生长曲线,荧光染色,荧光显微镜观察,流式细胞仪分析和实时定量PCR,检测雷公藤红素对前列腺癌细胞生长、凋亡及SENP1 mRNA表达水平的影响。结果：雷公藤红素能够显著抑制PCa细胞的生长,并且抑制作用呈剂量依赖性;雷公藤红素能够诱导PCa细胞凋亡,1μmol/L雷公藤红素处理24h诱导14.8%的PC-3细胞和23.2%的LNCaP细胞发生凋亡或死亡;雷公藤红素还能够降低PCa细胞中SENP1 mRNA水平,尤其是PC-3细胞。结论：雷公藤红素能够显著抑制PCa细胞的生长并诱导细胞凋亡,表明雷公藤红素具有抗前列腺癌作用,雷公藤红素能够降低PCa细胞中SENP1 mRNA水平,揭示雷公藤红素可能通过SENP1相关信号通路达到抗前列腺癌作用。     

雷公藤红素对肝叶部分切除老年小鼠术后认知功能及海马Tau蛋白磷酸化的影响

目的：观察肝叶部分切除术后老年小鼠认知功能的变化及海马内Tau蛋白磷酸化位点Ser396、Ser199/202的变化和雷公藤红素对老年小鼠术后认知功能障碍时的神经保护作用。方法：C57BL/6老年小鼠随机分为3组（n=48）：假手术组（S组）、肝叶部分切除术组（O组）和肝叶部分切除术加雷公藤红素组（C组）。每组再根据不同时间点分为12h、1d、3d和7d4个亚组,每个时间点12只动物。三组小鼠采用Mor-ris水迷宫检测小鼠认知功能,免疫组织化学法和Western blotting方法检测各组小鼠海马Tau蛋白磷酸化的变化。结果：与S组同时点比较,O组术后3d和7d时水迷宫成绩下降（P〈0.05）;C组术后3d和7d水迷宫成绩好于O组（P〈0.05）,而与S组接近（P〉0.05）。免疫组织化学染色阳性数和Western blotting显示O组Tau蛋白磷酸化位点Ser396、Ser199/202表达多于S组（P〈0.01）,而C组表达少于O组（P〈0.01）,与S组接近（P〉0.05）。结论：雷公藤红素对老年小鼠肝叶部分切除术造成的认知功能障碍具有明显的保护作用,其机制可能与抑制Tau蛋白异常磷酸化有关。