15,16-Dihydrotanshinone I from the Functional Food Salvia miltiorrhiza Exhibits Anticancer Activity in Human HL-60 Leukemia Cells: in Vitro and in Vivo Studies

15,16-Dihydrotanshinone I (DHTS) is extracted from Salvia miltiorrhiza Bunge which is a functional food in Asia. In this study, we investigated the apoptotic effect of DHTS on the human acute myeloid leukemia (AML) type III HL-60 cell line. We found that treatment with 1.5 μg/mL DHTS increased proapoptotic Bax and Bad protein expressions and activated caspases-3, -8, and -9, thus leading to poly ADP ribose polymerase (PARP) cleavage and resulting in cell apoptosis. DHTS induced sustained c-Jun N-terminal kinase (JNK) phosphorylation and Fas ligand (FasL) expression. The anti-Fas blocking antibody reversed the DHTS-induced cell death, and the JNK-specific inhibitor, SP600125, inhibited DHTS-induced caspase-3, -8, -9, and PARP cleavage. In a xenograft nude mice model, 25 mg/kg DHTS showed a great effect in attenuating HL-60 tumor growth. Taken together, these results suggest that DHTS can induce HL-60 cell apoptosis in vitro and inhibit HL-60 cell growth in vivo; the underlying mechanisms might be mediated through activation of the JNK and FasL signal pathways.     

Importance of Kupffer Cells in the Development of Acute Liver Injuries in Mice

Kupffer cells reside within the liver sinusoid and serve as gatekeepers. They produce pro- and anti-inflammatory cytokines and other biologically important molecules upon the engagement of pattern recognition receptors such as Toll-like receptors. Kupffer cell-ablated mice established by in vivo treatment with clodronate liposomes have revealed many important features of Kupffer cells. In this paper, we review the importance of Kupffer cells in murine acute liver injuries and focus on the following two models: lipopolysaccharide (LPS)-induced liver injury, which is induced by priming with Propionibacterium acnes and subsequent challenge with LPS, and hypercoagulability-mediated acute liver failure such as that in concanavalin A (Con A)-induced hepatitis. Kupffer cells are required for LPS sensitization induced by P. acnes and are a major cellular source of interleukin-18, which induces acute liver injury following LPS challenge. Kupffer cells contribute to Con A-induced acute liver failure by initiating pathogenic, intrasinusoidal thrombosis in collaboration with sinusoidal endothelial cells. The mechanisms underlying these models may shed light on human liver injuries induced by various etiologies such as viral infection and/or abnormal metabolism.     

ILLLI对银屑病患者外周血CD4+T细胞Fas蛋白表达的影响

目的:探索ILLLI治疗对银屑病患者外周血CD4+T细胞Fas蛋白表达的影响。方法:取患者及正常对照组静脉血10ml,肝素抗凝,等份分别加入4支试管中,1支为未照射组,3支照射组分为30、60和120分钟,以输出功率5mW的氦氖激光照射,并进行荧光染色和流式细胞分析。结果:照射前银屑病组表达Fas蛋白的CD4+T细胞百分比为21.4±3.1％,较正常对照组的16.8±2.1％显著增加,差异有显著性,p<0.05。正常对照组在激光照射30分钟、60分钟和120分钟,表达Fas蛋白的CD4+T细胞百分比分别为18.0±5.3％、16.4±5.2％和19.8±6.7％,与照射前差异无显著性,p>0.05;而患者组在激光照射30分钟、60分钟和120分钟时,表达Fas蛋白的CD4+T细胞百分比分别为24.3±2.8％、28.3±5.3％和30.5±7.9％,可以看出,随着照射时间的延长,表达Fas蛋白的CD4+T细胞的数量不断增加,60分钟和120分钟,表达Fas蛋白的CD4+T细胞百分比较照射前增加显著,有统计学差异,p<0.05。结论:银屑病的发病机制可能与细胞凋亡失控等多种因素有关。激光治疗银屑病的机制可能与它下调IL—8水平、上调TNF—(表达、纠正患者的凋亡失控状态有关。     

Modulation of cell growth and apoptosis response in human prostate cancer cells supplemented with tocotrienols

Previous studies have shown that tocotrienols are powerful growth inhibitors and potent inducers of apoptosis in human breast cancer cells. The objective of the current study was to examine effects of tocotrienols on apoptotic signals in androgen‐independent PC‐3 human prostate cancer cells. We investigated the effects of the tocotrienol‐rich fraction (TRF) from palm oil, α‐tocopherol (αT), α‐tocotrienol (αT₃), γ‐tocotrienol (γT₃) and δ‐tocotrienol (δT₃) on PC‐3 cell growth. TRF inhibited PC‐3 growth with a nonlinear response, with complete growth suppression at 10 µg/mL. δT₃ and γT₃ showed complete cell inhibition at 8 µg/mL whilst αT had no effect. δT₃ and γT₃ showed the most promise in the cell growth assays, and all subsequent experiments were performed with δT₃, TRF and αT. TRF and δT₃ at 8 µg/mL induced apoptosis in PC‐3 cells after 48 h of treatment. In addition, TRF and δT₃ treatments were able to affect the cell cycle, with accumulation in the S phase, G2 phase block and increases in SubG1 by 72 h. We then proceeded to investigate the expression levels of Fas receptor and Fas ligand, caspase 8, caspase 3 and bax in PC‐3 cells following treatment with tocotrienols using real‐time PCR and Western blot methods. TRF and δT₃ at 8 µg/mL increased Fas ligand expression levels by 368 and 456%, respectively, after 24 h and Fas receptor expression levels by 210% and 356%, respectively, after 48 h. TRF‐ and δT₃‐treated PC‐3 cells overexpressed caspase 8 and bax protein after 24 h, and caspase 3 after 48 h. In conclusion, tocotrienols are able to induce apoptosis and cell cycle arrest in PC‐3 cells, with increased expression of Fas receptor, Fas ligand, caspase 8, caspase 3 and bax, suggesting a potential role in chemoprevention of prostate cancer.     

PDTC antagonized polysaccharide-induced apoptosis in MCF-7 cells through a caspase-8 mediated Fas pathway

Cartilage polysaccharide (PS) was extracted from porcine cartilage to investigate its anti-tumor effect. In this study, we showed that PS induced apoptosis in MCF-7 breast cancer cells by activating the caspase-8 mediated Fas signaling pathway. The PS inhibited MCF-7 cell proliferation in a dose-dependent and time-dependent manner, and had no toxic effect on normal cells such as NIH-3T3 cells. An increase of Fas protein expression was first located on the nuclear membrane and then on the outside membrane. Cleaved caspase-8 was activated after PS treatment. NF-κB activation was not involved in the PS induced apoptosis, but the NF-κB inhibitor PDTC may act as a PS antagonist to reduce the apoptosis rate and Fas protein expression in PS-induced apoptosis in MCF-7 cells.     

微胶囊固定化培养重组盘基网柄菌提高人类可溶性Fas配体的表达

首先研究了应用于液芯羧甲基纤维素钠-海藻酸钙(CMC-ALG)微胶囊制备中的各种化学组分对重组盘基网柄菌生长的影响,然后考察不同组分浓度制备而成的各种CMC-ALG微胶囊内重组盘基网柄菌的生长情况,从而得到较适的微胶囊制备的组分配比(CMC 12 g·L-1,SA 8 g·L-1,CaCl2 100 g·L-1).结果表明,在以上较适条件下制备的微胶囊内重组盘基网柄菌的生长得到了极大的改善,最大的细胞密度比游离培养时提高了4倍,达到了8.0×107 mL-1;相应的人类可溶性Fas配体(FasL)的表达水平也提高了1.5倍,达到了315 μg·L-1.最后,开展了微胶囊化重组盘基网柄菌的二次重复发酵FasL的研究,结果表明,经过二次重复批次培养,最大细胞密度可达到1.24×108 mL-1,为游离培养的8～10倍,而且FasL的表达水平还能维持高水平(280 μg·L-1),为游离培养时的2倍.     

CD28和Fas在辐射所致T细胞凋亡中作用的研究

采用双标记直接免疫荧光－流式细胞仪分析技术,测定了不同剂量γ射线照射后T细胞CD28分子和Fas受体（CD95）表达的变化。用乙醇抽提法－流式细胞仪分析技术和JAM检测技术,研究了不同剂量γ射线照射、CD28单抗和IL－18处理后T细胞凋亡的情况。结果显示,正常人淋巴细胞用不同剂量γ射线照射后,CD3^ T细胞中CD28的表达随着剂量的增加而下降,T细胞凋亡的增加并与Fas表达上升有关。照射前用CD28单抗和IL－18处理细胞后,Fas表达下调,细胞DNA断裂明显减小。表明γ射线可影响T细胞CD28受体分子的表达及其信号转导,加速T细胞凋亡的进程,而CD28单抗和IL－18对辐射所致T细胞凋亡有一定的拮抗作用。     

川芎嗪对儿茶酚胺诱导大鼠心肌损伤时Bcl-2和Fas 蛋白表达的影响

目的: 观察川芎嗪对儿茶酚胺诱导大鼠心肌损伤时心肌细胞Bcl-2 和Fas 蛋白表达的影响。方法: 采用皮下多点注射异丙肾上腺素(ISO,5 mg·kg^-1) 每天1 次, 连续3 d, 诱导大鼠心肌损伤,采用免疫组化方法检测Bcl-2 和Fas 蛋白的水平, 并利用图像分析系统分析蛋白阳性表达区域平均光密度值。结果: 损伤组和川芎嗪保护组心肌细胞Bcl-2蛋白表达较对照组均显著升高(P<0.01), 损伤组Fas 蛋白水平较对照组也显著升高(P<0.01), 川芎嗪保护组的Fas 蛋白水平较损伤组显著下降(P<0.01), 且Bcl-2 Fas 比值也较损伤组和对照组明显增加。川芎嗪保护组的病理损伤程度明显低于损伤组。结论: 川芎嗪可抑制儿茶酚胺诱导的心肌损伤时心肌细胞中促凋亡基因Fas 的表达, 提高心肌细胞中Bcl-2 Fas 比值。     

参麦注射液对肺缺血-再灌注损伤时Fas/FasL 表达的影响

目的 探讨参麦注射液对肺缺血-再灌注损伤(PIRI) 时Fas/FasL 配体(Fas/FasL) 表达的影响。方法 采用在体兔单肺原位缺血-再灌注模型。实验兔30 只, 随机分为假手术对照组(sham,n=10) 、肺缺血-再灌注组(I-R, n=10) 和肺缺血-再灌注加参麦注射液组(SM, n=10)。分别于再灌注3 h 取左肺组织, 观察Fas/FasL mRNA 定位表达、凋亡指数(AI) 、肺组织湿干重比(W D) 、肺损伤组织学定量评价指标(IQA) 及光镜、电镜下的组织形态学改变。结果 SM 组Fas/FasL mRNA在肺小动脉内(外) 膜、肺小静脉内膜、肺泡上皮及肺支气管上皮呈弱阳性表达, 明显低于I-R 组(P<0.05);AI 、W D 和IQA 值显著低于I-R 组(P <0.01 和P <0.05);肺组织形态学异常改变程度减轻。结论 参麦注射液可下调肺组织Fas/FasLmRNA 的表达而减轻细胞凋亡, 对PIRI 发挥积极的防治作用。