Antiprotozoal activity of Neurolaena lobata.

Extracts, fractions and sesquiterpene lactones from Neurolaena lobata (L.) R. Br. (Asteraceae), a traditional medicinal plant from Guatemala, were tested in vitro against Leishmania spp. promastigotes, Trypanosoma cruzi trypomastigotes and epimastigotes and Trichomonas vaginalis trophozoites. The ethanol extract inhibited the parasite growth of L. mexicana, T. cruzi and T. vaginalis significantly. The pure germacranolides 1 and a mixture of 2 and 3, isolated from the ethonal extract, were highly active against L. mexicana and T. cruzi.     

霉滴洗剂体外抗阴道滴虫实验研究

利用不同浓度的霉滴洗剂药液分别与不同虫株的阴道滴虫做体外实验，观察其体外抗阴道滴虫效果。作用２４、４８ｈ后观察结果显示，该中药方剂对不同虫株的阴道滴虫的作用无明显差异。最低杀虫浓度为１∶４。以上结果表明霉滴洗剂在体外有较强的抗滴虫作用。在滴虫性阴道炎的临床治疗中有重要意义     

7,7'-Dimethoxyagastisflavone-induced apoptotic or autophagic cell death in different cancer cells

7,7'-Dimethoxyagastisflavone (DMGF), a biflavonoid isolated from the needles of Taxus × media cv. Hicksii, was evaluated for its antiproliferative and antineoplastic effects in three human cancer cell lines. Interestingly, DMGF caused cell death via different pathways in different cancer cells. DMGF induced apoptosis, activated caspase-3 activity and changed the mitochondrial membrane potential in HT-29 human colon cancer cells. However, the apoptotic pathway is not the major pathway involved in DMGF-induced cell death in A549 human lung cancer cells and HepG2 human hepatoma cells. Treatment with 3-MA, an inhibitor of autophagy, significantly decreased DMGF-induced cell death in HepG2 and A549 cells, but did not affect DMGF-induced cell death in HT-29 cells. Following DMGF treatment, the HepG2 cells increased expression of LC3B-II, a marker used to monitor autophagy in cells. Thus, DMGF induced apoptotic cell death in HT-29 cells, triggered both apoptotic and autophagic death in A549 cells and induced autophagic cell death in HepG2 cells.     

Genistein protects against amyloid‐beta‐induced toxicity in SH‐SY5Y cells by regulation of Akt and Tau phosphorylation

Alzheimer's disease is a neurodegenerative disorder characterized by extracellular deposition of amyloid‐β (Aβ) peptide and hyperphosphorylation of Tau protein, which ultimately leads to the formation of intracellular neurofibrillary tangles and cell death. Increasing evidence indicates that genistein, a soy isoflavone, has neuroprotective effects against Aβ‐induced toxicity. However, the molecular mechanisms involved in its neuroprotection are not well understood. In this study, we have established a neuronal damage model using retinoic‐acid differentiated SH‐SY5Y cells treated with different concentrations of Aβ_25–35 to investigate the effect of genistein against Aβ‐induced cell death and the possible involvement of protein kinase B (PKB, also termed Akt), glycogen synthase kinase 3β (GSK‐3β), and Tau as an underlying mechanism to this neuroprotection. Differentiated SH‐SY5Y cells were pre‐treated for 24 hr with genistein (1 and 10 nM) and exposed to Aβ_25–35 (25 μM), and we found that genistein partially inhibited Aβ induced cell death, primarily apoptosis. Furthermore, the protective effect of genistein was associated with the inhibition of Aβ‐induced Akt inactivation and Tau hyperphosphorylation. These findings reinforce the neuroprotective effects of genistein against Aβ toxicity and provide evidence that its mechanism may involve regulation of Akt and Tau proteins.     

Bisabololoxide A,One of the Constituents in German Chamomile Extract,Attenuates Cell Death Induced by Calcium Overload

Bisabololoxide A (BSBO), main constituents in German chamomile extract, is responsible for antipruritic effect. In previous study, the incubation with 30–100 μM BSBO for 24 h exerted cytotoxic and proapoptotic effects on rat thymocytes. To further characterize BSBO cytotoxicity, the effect on the cells suffering from calcium overload by calcium ionophore A23187 was examined. A23187 induced Ca²⁺‐dependent cell death. Contrary to our expectation, 1–10 μM BSBO inhibited A23187‐induced increase in cell lethality of rat thymocytes. BSBO attenuated A23187‐induced increases in populations of shrunken living cells, phosphatidylserine‐exposed living cells, and dead cells, without affecting the increase in intracellular Ca²⁺ concentration and the Ca²⁺‐dependent hyperpolarization. The effect of BSBO on A23187‐treated cells may be unique because the activation of Ca²⁺‐dependent K⁺ channels is required for cell shrinkage, externalization of phosphatidylserine, and cell death in some cells. The cell death induced by A23187 was not inhibited by Z‐VAD‐FMK, a pan‐inhibitor of caspases. Thus, the cell death may be a necrosis with some features observed during an early stage of apoptosis. These results suggest that BSBO at low micromolar concentrations is cytoprotective against calcium overload. Copyright © 2013 John Wiley & Sons, Ltd.     

Protective effect of a Chrysanthemum indicum containing formulation in cadmium-induced ototoxicity

Chungshinchongyitang (CSCYT) is an herbal drug formula containing Chrysanthemum indicum and 13 other herbs used for treating auditory diseases. Irreversible hearing loss is a characteristic effect of a number of heavy metals. Cadmium (Cd(2+)) is an environmental contaminant that causes a variety of adverse effects. In the present study, we investigate the protective effects of CSCYT against Cd(2+) induced ototoxicity in vitro and ex vivo. The findings of this study show that CSCYT prevents the destruction of hair cell arrays induced by Cd(2+) in the rat organ of Corti primary explants. CSCYT inhibited cell death, release of cytochrome c and generation of reactive oxygen species induced by Cd(2+) in HEI-OC1 auditory cell line. In addition, we also demonstrated that CSCYT exerted its effect by modulating of apoptosis via the caspase-3 activation and extracellular signal-regulated kinase activation. These results are expected to improve the understanding of the pharmacological mechanism of CSCYT and aid in the development of potential therapeutic strategies against ototoxicity.     

20S-protopanaxadiol-induced programmed cell death in glioma cells through caspase-dependent and -independent pathways

20S-Protopanaxadiol (1) is an aglycon metabolic derivative of the protopanaxadiol-type ginseng saponins. In the present study, 1 was used to induce cytotoxicity for two human glioma cell lines, SF188 and U87MG. For the SF188 cells, 1 activated caspases-3, -8, -7, and -9 within 3 h and induced rapid apoptosis, which could be partially inhibited by a general caspase blocker and completely abolished when the caspase blocker was used in combination with an antioxidant. Compound 1 also induced cell death in U87MG cells but did not activate any caspases in these cells. Monodansylcadaverine staining showed that 1 induced dramatic autophagy in both cell lines. Elevated levels of superoxide anion in both cells and reduced levels of phosphorylated Akt in U87MG cells were also demonstrated. These results showed that 20S-protopanaxadiol (1) induces different forms of programmed cell death, including both typical apoptosis and autophagy through both caspase-dependent and -independent mechanisms.     

Coriolus versicolor‐derived protein‐bound polysaccharides trigger the caspase‐independent cell death pathway in amelanotic but not melanotic melanoma cells

We have investigated the potential cell death mechanism promoted by Coriolus versicolor fungus‐derived protein‐bound polysaccharides (PBPs) in melanoma cells. Knowing that melanogenesis has the potential to affect the tumor behavior and melanoma therapy outcome, the cytotoxic effects of PBPs were evaluated in human SKMel‐188 melanoma cell line, whose phenotype, amelanotic versus pigmented, depends on the concentration of melanin precursors in the culture medium. Our results showed that inhibitory effect of PBPs (100 and 200 μg/ml) towards melanoma cells is inversely associated with the pigmentation level. This cytotoxicity induced in nonpigmented melanoma cells by PBPs was caspase‐independent; however, it was accompanied by an increased intracellular reactive oxygen species (ROS) generation. The ROS production was controlled by c‐Jun N‐terminal kinase (JNK) because SP600125, a JNK inhibitor, significantly reduced ROS generation and protected cells against PBPs‐induced death. We also found that PBPs‐induced lactate dehydrogenase release in amelanotic melanoma cells was abolished by co‐treatment with receptor‐interacting serine/threonine‐protein kinase 1 inhibitor, implying engagement of this kinase in PBPs‐induced death pathway. The results suggest that PBPs induce an alternative programmed cell death, regulated by receptor‐interacting protein‐1 and ROS and that this process is modified by melanin content in melanoma cells. These findings are remarkable when considering the use of commercially available Coriolus versicolor by patients who suffer from melanoma cancer.     

Zingerone activates VMAT2 during MPP+‐induced Cell Death

Parkinson's disease (PD) is characterized by a progressive and selective loss of dopaminergic neurons in the substantia nigra pars compacta (SNpc) and striatum. 1‐Methyl‐4‐phenyl‐1,2,3,6‐tetrahydropyridine (MPTP) is used to produce an animal model for PD, and it is converted to 1‐methyl‐4‐phenylpyridine (MPP⁺) in animals. MPP⁺ accumulation leads to neuronal cell death. Vesicular monoamine transporter 2 (VMAT2) regulates the accumulation of monoamine neurotransmitters into synaptic vesicles and is involved in neuroprotection against neurotoxin‐induced cell death. Recently, zingerone has been reported to reduce oxidative stress and inhibit inflammation. Therefore, we examined the effect of zingerone on neuronal cell death in a PD model. In an MPP⁺ and MPTP‐mediated PD model, neuronal cell survival was increased by zingerone without modifying neuroinflammation or reactive oxygen species generation. Zingerone also induced ERK activation and VMAT2 expression, leading to the attenuation of MPP⁺‐induced neuronal cell death. Our current results suggest that zingerone has a neuroprotective effect in a PD model. Copyright © 2015 John Wiley & Sons, Ltd.     

洁而舒洗剂药效学研究

目的观察洁而舒洗剂对动物炎症、瘙痒、细菌及阴道毛滴虫感染的影响。方法采用体外试管法测定洁而舒洗剂最小抑菌浓度；以化学刺激法观察洁而舒洗剂对小鼠和大鼠肿胀反对豚鼠痒闾的影响。结果洁而舒洗剂对葡萄球菌等多种细菌及滴虫均具有不同程度的抑制作用；能明显抑制小鼠耳廓肿胀和大鼠足跖肿胀；其作用呈剂量依赖性．能显著提高豚鼠对组胺的致痒阈。结论洁而舒洗剂具有抗炎、抑菌和杀虫及止痒作用。     

促红细胞生成素对脑缺氧的保护作用

 目的在细胞水平探讨促红细胞生成素(EPO)对缺氧诱发的神经元损伤的改善作用及缺糖缺氧诱发的脑片损伤的改善作用。方法　采用原代新生大鼠皮层培养和小鼠脑片的方法,分别以形态学和二苯基四氮唑溴盐(MTT)及2,3,5-三苯基氯化四氮唑(TTC)染色为指标观察神经元及脑片的损伤和药物的改善作用。结果EPO在10^-11,10^-12mol·L^-1能显著改善缺氧24 h引起的培养神经元死亡,其在10^-9mol·L^-1能改善缺氧45min或缺糖缺氧15min引起的脑片梗死。结论EPO对缺氧诱发的神经元死亡和缺糖缺氧诱发的脑片梗死具有保护作用。     

Inhibition of cancer cell growth by crude extract and the phenolics of Terminalia chebula retz. fruit

A 70% methanol extract of Terminalia chebula fruit, was studied for its effects on growth in several malignant cell lines including a human (MCF-7) and mouse (S115) breast cancer cell line, a human osteosarcoma cell line (HOS-1), a human prostate cancer cell line (PC-3) and a non-tumorigenic, immortalized human prostate cell line (PNT1A) using assays for proliferation ([(3)H]-thymidine incorporation and coulter counting), cell viability (ATP determination) and cell death (flow cytometry and Hoechst DNA staining). In all cell lines studied, the extract decreased cell viability, inhibited cell proliferation, and induced cell death in a dose dependent manner. Flow cytometry and other analyses showed that some apoptosis was induced by the extract at lower concentrations, but at higher concentrations, necrosis was the major mechanism of cell death. ATP assay guided chromatographic fractionation of the extract yielded ellagic acid, 2,4-chebulyl-beta-D-glucopyranose (a new natural product), and chebulinic acid which were tested by ATP assay on HOS-1 cell line in comparison to three known antigrowth phenolics of Terminalia, gallic acid, ethyl gallate, luteolin, and tannic acid. Chebulinic acid (IC(50) = 53.2 microM +/- 0.16) > tannic acid (IC(50) = 59.0 microg/ml +/- 0.19) > and ellagic acid (IC(50) = 78.5 microM +/- 0.24), were the most growth inhibitory phenolics of T. chebula fruit in our study.     

Yokukansan, a kampo medicine, protects against glutamate cytotoxicity due to oxidative stress in PC12 cells

Aim of the study

Yokukansan is a traditional Japanese medicine consisted of seven medicinal herbs and has been used for treatment of neurosis, insomnia, and behavioral and psychological symptoms of dementia in Japan. The aim of the present study is to clarify the active compounds responsible for the protective effect of yokukansan against glutamate-induced cytotoxicity in PC12 cells.

Materials and methods

PC12 cells which is a tool for selective evaluation of test substances against oxidative stress was used in the present study. The cell survival rates or glutathione (GSH) levels were evaluated by a MTT reduction assay or GSH assay based on the GSH reductase enzymatic recycling method, respectively.

Results

Glutamate (1-17.5 mM) induced cell death of PC12 cells in a concentration- dependent manner. Yokukansan (125-500 μg/ml) inhibited the glutamate-induced PC12 cell death. When the effects of extracts of the seven constituent herbs in yokukansan on the cell death were examined, Uncaria thorn was found to have the highest potency in the protection. To clarify the active compounds in Uncaria thorn, the effects of seven alkaloids (rhynchophylline, isorhynchophylline, corynoxeine, isocorynoxeine, hirsutine, hirsuteine, and geissoschizine methyl ether) on the cell death were further examined. The protective effects were found in hirsutine, hirsuteine, and geissoschizine methyl ether, which also ameliorated the glutamate-induced decrease in GSH levels.

Conclusion

These results suggest that yokukansan protects against PC12 cell death induced by glutamate-mediated oxidative stress, i.e., reduction of intracellular GSH level, and the effect may be mainly attributed to a synergistic effect of the hirsutine, hirsuteine, and geissoschizine methyl ether in Uncaria thorn.     

Neuroprotective effect of honokiol and magnolol, compounds from Magnolia officinalis, on beta-amyloid-induced toxicity in PC12 cells

Amyloid β peptide (Aβ) induced toxicity is a well-established pathway of neuronal cell death which might play a role in Alzheimer's disease. In this regard, the toxic effect of Aβ on a cultured Aβ-sensitive neuronal cell line was used as a primary screening tool for potential anti-Alzheimer's therapeutic agents. The effects of nine pure compounds (vitamin E, α-asarone, salidroside, baicolin, magnolol, gastrodin, bilobalide, honokiol and β-asarone) from selected Chinese herbs on neuronal cell death induced by Aβ in NGF-differentiated PC12 cells were examined. Only two of the studied compounds, honokiol and magnolol, significantly decreased Aβ-induced cell death. Further experiments indicated that their neuroprotective effects are possibly mediated through reduced ROS production as well as suppression of intracellular calcium elevation and inhibition of caspase-3 activity. The results provide for the first time a scientific rationale for the clinical use of honokiol and magnolol in the treatment of Alzheimer's disease.     

Protection of brain cells against AMPA-induced damage by Asiasari Radix extracts

We determined whether Asiasari Radix (AR) extracts have protective actions in brain cells. Methanol extracts of Asiasari Radix (fraction 1) have significant inhibitory effects on the AMPA-induced rat cortical depolarization in the grease gap assay. In differentiated PC12 cells, it almost completely protected against AMPA-induced cell death. In addition, it had some protective actions in C6 glial cells death induced by AMPA. The methanol extracts (fraction 1) of AR were subsequently fractionated into chloroform-(fraction 2), chloroform/methanol-(3:1) (fraction 3), methanol-soluble (fraction 4) and methanol-insoluble, water-soluble fractions (fraction 5). Among these, fraction 4 had the strongest inhibitory effects against AMPA-induced cell death in the PC 12 cells and also dramatically inhibited AMPA-induced depolarization of rat brain cortex in the grease gap assay. Interestingly, fraction 4 blocked the Zn-induced oxidative damages in C6 glial cells.     

Brucea javanica fruit induces cytotoxicity and apoptosis in pancreatic adenocarcinoma cell lines

Brucea javanica fruit is thought to have anticancer properties in Chinese medicine and its extract has been shown to possess antiproliferative and pro-apoptotic activities on human carcinoma cells. In the present study we demonstrated for the first time that Fructus Bruceae extract exhibited cytotoxic effects on the three pancreatic adenocarcinoma cell lines, PANC-1, SW1990 and CAPAN-1; the effects were comparable to those exhibited by camptothecin in our culture system. In addition, Fructus Bruceae extract induced fragmentation of genomic DNA, as evidenced by Hoechst staining and the cell death detection ELISA(PLUS) assay. Western blot analysis further showed down-regulation of pro-caspase 3 protein expression, indicating that the observed cytotoxic effects of the extract were associated with induction of apoptosis. These findings are not only significant in the development of traditional Chinese medicine as an alternative treatment for pancreatic cancer, but also in the elucidation of the potential mechanism(s) of Fructus Bruceae extract in cancer therapy.     

Synergistic effect of green tea polyphenols on their protection against FK506-induced cytotoxicity in renal cells

FK506 (tacrolimus) is a widely used immunosuppressant first employed in the management of rejection in organ transplantation, but now used for autoimmune disease. However, the nephrotoxicity induced by FK506 remains a serious clinical problem. We previously demonstrated that FK506 caused a significant increase in apoptosis of LLC-PK1 cells, a porcine proximal tubule cell line, but the addition of green tea extract and its polyphenolic components suppressed the cell death. Here, we examined the synergistic effect of tea polyphenols on the protection of FK506-induced cell death. The combined treatment with 5 microM (-)-epigallocatechin-gallate (EGCG) and 5 microM of (+)-catechin (C), (-)-epicatechin (EC), (-)-epigallocatechin (EGC) or (-)-epicatechin-gallate (ECG) reduced FK506-induced cytotoxicity in LLC-PK1. Similarly, the combined treatment with 5 microM EGC and 5 microM of C, EC, EGCG or ECG also reduced the cytotoxicity. These results showed that the co-treatments with EGCG and EGC, EGCG or ECG, and EGC and ECG have stronger synergistic effects on the protection of FK506-induced cell death. Furthermore, the combined treatment of EGCG (5 microM) and EGC (5 microM) showed a significant time-dependent suppression of the increased intracellular ROS levels 15 min after the addition of FK506, as well as on caspase activation. The results of these synergistic effects of the constituents of green tea extract suggest that its protective effects may reside in more than just one of its constituent.     

11种中药体外灭阴道毛滴虫

为了开发新的中药用于治疗阴道毛滴虫，对１１种具有杀虫、消炎作用的中药进行了体外灭阴道毛滴虫实验，并对有效药物交叉配伍组成复主，分别进行体外灭虫实验，结果表明：单剂以常山、花椒、白头翁、青蒿、仙鹤草杀虫效果较好，复方以常山＋花椒和白头翁＋青蒿灭虫效果最好，提示以上药物可作为抗滴虫新药应用于临床。