藿苓益智方对阿尔茨海默病模型小鼠神经细胞凋亡线粒体通路的作用研究

目的:探讨藿苓益智方对阿尔茨海默病线粒体通路神经细胞凋亡的调控作用。方法:采用Aβ的活性片段Aβ_25-35诱导PC12细胞凋亡建立阿尔茨海默病的细胞模型。本实验设置六个组,即对照组、Aβ_25-35组、Aβ_25-35+盐酸多奈哌齐组、Aβ_25-35+藿苓益智方提取物(50、100、200 μg/ml)组。利用流式细胞仪、荧光酶标仪和透射电镜等技术,检测霍苓益智方提取物对阿尔茨海默病细胞模型中细胞存活率、神经细胞凋亡率、线粒体超微结构的变化。结果:藿苓益智方提取物50 μg/ml组的细胞存活率与Aβ_25-35组相比有一定的增高(P<0.05); 盐酸多奈哌齐组和藿苓益智方提取物(100、200 μg/ml)组与Aβ_25-35组相比,细胞存活率显著升高(均P<0.01),但藿苓益智方提取物200 μg/ml组升高的更明显; 藿苓益智方低、中、高剂量+A_25-35组细胞凋亡率则逐步下降,其中中、高剂量组与Aβ_25-35组相较差异均具有统计学意义(均P<0.01); 藿苓益智方低、中、高剂量+Aβ_25-35组对细胞核的损害程度上,经治疗以后逐步缓解,且呈剂量依赖性趋势。结论:藿苓益智方对阿尔茨海默病的治疗中可对Aβ诱导的PC12细胞凋亡具有明显的抑制作用,改善细胞存活率,减少线粒体细胞凋亡。     

Pycnogenol Cytotoxicity in Pancreatic INS‐1E β cells Induced by Calcium Dysregulation

Natural standardized flavonoid extract from the bark of Pinus pinaster, Pycnogenol (Pyc), was recently found to decrease intensively the activity of sarcoplasmic reticulum Ca²⁺‐ATPase of rabbit skeletal muscle (SERCA1). On the basis of this inhibitory effect in a cell‐free system and similarities of SERCA1 to its other isoforms, proapoptotic properties of Pyc may be expected in cellular systems. Pycnogenol (40–100 μg/mL) induced a concentration‐dependent decrease of the viability of pancreatic INS‐1E β cells associated with induction of apoptosis. In addition, intracellular Ca²⁺ level increase was found along with reduction of protein expression level of SERCA2b and impairment of insulin secretion by β cells. These facts indicate that Pyc may induce apoptosis by impairment of calcium homeostasis. Copyright © 2017 John Wiley & Sons, Ltd.     

Thymoquinone Crosstalks with DR5 to Sensitize TRAIL Resistance and Stimulate ROS-Mediated Cancer Apoptosis

Objective: Cancer treatment using a targeted inducer of apoptosis like tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) faced the obstacle of resistance, thus providing a plus drug like Thymoquinone (TQ) could be of great interest to tackle breast cancer cells. The aim of the present work is to examine the genetic modulation impacts of the TRAIL receptors and apoptotic markers upon the combinatorial remedy of TRAIL plus TQ on human breast cancer cell lines. Methods: To achieve this rationale, the protein content-based cytotoxicity using SRB assay, as well as the genetic expressions of the TRAIL receptors (DR4 and DR5) and apoptotic markers (Bcl-2, Cas-8, and FADD) using real time qRT-PCR technique were preceded against breast cancer MCF-7 and MDA-MB-231 cancerous cell lines. Results: The current study showed that the combination therapy of TQ+TRAIL significantly inhibited the protein content-based proliferation of MDA-MB-231 cells more than MCF-7 cells. The synergistic effect of them significantly up-regulated the genetic expressions of DR4, DR5, Cas-8, and FADD genes and inhibited the genetic expression of the Bcl-2 gene in the proposed cell lines treated for 24 h. The induction of the apoptotic genes using the combined therapy was stimulated by the elevation of the reactive oxygen species (ROS); nitric oxide (NO) and malondialdehyde (MDA) levels. Conclusions: The synergistic influence between TQ which induced the DR5 and TRAIL, facilitating the connection between TRAIL and its receptors on the cancerous cell membrane. Hence, the proposed combination therapy induced the ROS-mediated apoptotic stimulus.     

Combined inhibition of Bcl-2/Bcl-xL and Usp9X/Bag3 overcomes apoptotic resistance in glioblastoma in vitro and in vivo

Despite great efforts taken to advance therapeutic measures for patients with glioblastoma, the clinical prognosis remains grim. The antiapoptotic Bcl-2 family protein Mcl-1 is overexpressed in glioblastoma and represents an important resistance factor to the BH-3 mimetic ABT263.In this study, we show that combined treatment with ABT263 and GX15-070 overcomes apoptotic resistance in established glioblastoma cell lines, glioma stem-like cells and primary cultures. Moreover, this treatment regimen also proves to be advantageous in vivo. On the molecular level, GX15-070 enhanced apoptosis by posttranslational down-regulation of the deubiquitinase, Usp9X, and the chaperone Bag3, leading to a sustained depletion of Mcl-1 protein levels. Moreover, knock-down of Usp9X or Bag3 depleted endogenous Mcl-1 protein levels and in turn enhanced apoptosis induced through Bcl-2/Bcl-xL inhibition.In conclusion, combined treatment with ABT263 and GX15-070 results in a significantly enhanced anti-cancer activity in vitro as well as in vivo in the setting of glioblastoma. Both drugs, ABT263 and GX15-070 have been evaluated in clinical studies which facilitates the translational aspect of taking this combinatorial approach to the clinical setting. Furthermore we present a novel mechanism by which GX15-070 counteracts Mcl-1 expression which may lay a foundation for a novel target in cancer therapy.     

Anticancer activities of the extract from Longkong (Lansium domesticum) young fruits

Context: “Longkong”(Lansium domesticum Corr., Family: Meliaceae) is a fruit found in the south of Thailand. This plant has been used in traditional medicines.

Objectives: To investigate the antiproliferative activities and the phytoconstituents of Longkong extracts.

Materials and methods: Cytotoxicity and apoptotic activity of 48 extracts were tested using the SRB assay and acridine orange (AO)/ethidium bromide (EB) staining, respectively. The extracts which gave the highest anticancer activity were selected to prepare the semipurified extracts and analysis for the constituents by gas chromatography–mass spectrometry (GC/MS).

Results: The highest percentage yield (59.38%) was from the cold water extract of Longkong ripe fruits (RFWC). The highest total phenolic and flavonoid contents were observed in cold and hot methanol extract of Longkong stalks (STMC and STMH). The hot and cold chloroform young Longkong fruit extracts (YFCH and YFCC) exhibited a cytotoxic effect (IC₅₀ < 1 mg/mL) against cancer cells. For apoptotic induction, YFCH demonstrated the highest activity against KB of 13.84 ± 4.21% at 0.5 mg/mL which was 0.88 and 1.35 times of cisplatin and 5-FU, respectively, while apoptotic cells in HT-29 were 8.68 ± 1.85% at 5 mg/mL, which was 0.61 and 1.43 times of cisplatin and 5-FU, respectively. YFCC showed the highest apoptotic effect against KB cells at 10.70 ± 2.15% at 0.5 mg/mL, which was 0.68 and 1.07 times of cisplatin and 5-FU, respectively. The major phytoconstituents in YFCH were hexadecanoic acid (11.53%) and ethyl oleate (10.58%).

Discussion and conclusion: The crude extracts of Longkong showed anticancer activities and may provide new lead compounds for the development of anticancer products.     

线粒体介导的细胞凋亡与男性不育的研究进展

随着世界工业化进程的加快,环境污染逐渐加重,男性不育的发病率呈全球性的增高趋势。疾病、毒物接触、电磁辐射、环境雌激素都可能是造成生育能力下降的诱因,但造成男性不育的机制尚不明确。精子质量低下是男性不育的根本原因,主要包括精子DNA损伤、精子活动力下降、精子密度降低等。线粒体损伤与精子细胞凋亡率升高常伴随以上原因出现。该文对线粒体介导的细胞凋亡与男性不育之间的关系进行简要综述。     

Cutaneous lesions of Kikuchi's disease: Evolution of histopathological findings

We report a 38‐year‐old male patient who presented with cutaneous lesions mimicking widespread discoid lupus erythematosus with high‐grade fever, arthralgia and lymphadenopathy. Additional lymph node and skin biopsies, however, revealed karyorrhectic debris without neutrophils and numerous CD68‐positive cells, a characteristic finding of Kikuchi's disease (KD). Comparing skin biopsies on different occasions, we could see different forms of histopathology. The histopathology of skin lesions of KD may vary during the clinical course, which may reflect the stage of the disease.     

Induction of Apoptosis in Nasal Polyp Fibroblasts by Glucocorticoids In Vitro

Objective—To examine the possible mechanisms underlying the therapeutic mode of action of glucocorticoids (GCs) in nasal polyposis.

Material and Methods—The effects of GCs on nasal polyps were firstly evaluated by examining the growth of fibroblasts derived from 10 nasal polyps in vitro. Subsequently, the ability of GCs to induce apoptotic cell death in fibroblasts was examined.

Results—Addition of betamethasone 21-phosphate (BET) at a concentration of > 1 × 10^?3 M to cell cultures inhibited cell growth in all cases examined. BET and dexamethasone 21-phosphate, but not testosterone or estradiol, caused apoptotic cell death in 2/10 nasal polyp fibroblasts, as assessed by agarose gel electrophoresis, when the cells were cultured with the agents for >96 h. The minimum concentration of agent needed to cause apoptosis was 1 × 10^?3 M, which is half of the recommended therapeutic dose.

Conclusion—The present findings suggest that topical application of GCs in nasal polyposis patients suppresses proliferation of fibroblasts in polyps and results in favorable modification of the clinical status of these patients.