NAD(P)H-quinone oxidoreductase-1 (NQO1) is a down-stream target gene of nuclear factor erythroid 2-related factor 2 (Nrf2), and performs diverse biological functions. Recently, NQO1 is recognized as an effective gene for the cytotoxic inserts with its diverse biological functions, which is focused on antioxidant properties. The aim of present study was to assess the impact of NQO1 knockdown on cytoprotection-related protein expression in cisplatin cytotoxicity by using small interfering (si) RNA targeted on NQO1 gene. Cytotoxicity of cisplatin on ACHN cells was assessed in a dose- and time-dependent manner after siScramble or siNQO1 treatment. After cisplatin treatment, cells were subjected to cell viability assay, western-blot analysis, and immunofluorescence study. The cell viability was decreased in the siNQO1 cells (50%) than the siScramble cells (70%) after 24 h of cisplatin (20 µM) treatment. Moreover, cytoprotection-related protein expressions were markedly suppressed in the siNQO1 cells after cisplatin treatment. The expression of Nrf2 and Klotho were decreased by 20% and 40%, respectively, of that in siScramble cells. Nrf2 and Klotho activation were also decreased in cisplatin treated siNQO1 cells, confirmed by cytoplasm-to-nuclear translocation. Our findings demonstrate that the increased cisplatin-induced cytotoxicity was accompanied by suppressed Nrf2 activation and Klotho expression in siNQO1 cells.
Cell Survival ; Cisplatin* ; Cytoprotection* ; Fluorescent Antibody Technique ; RNA

Animals ; Cytoprotection ; Humans ; Ischemic Preconditioning ; Reperfusion Injury ; prevention & control ; therapy

OBJECTIVE: A major limiting factor in human cancer chemotherapy is toxicity in normal cells and tissues. Our goal was to determine whether normal proliferating cells could be protected from chemotherapeutic agents by taking advantage of the differential drug sensitivity of cell cycle G1 checkpoint in normal and cancer cells. METHODS: Normal peripheral blood mononuclear cells (PBMC) and ovarian cancer cell lines (OVCAR- 3 and SKOV-3) were initially treated with 10 nM of staurosporine for 48 hours. After removal of staurosporine contained media, both PBMC and ovarian cancer cells were treated with 20 nM of paclitaxel for 24 hours. Cells were then allowed to recover in drug-free medium for 4 days. The DNA contents and cell cycle changes were detected by FACScan flow cytometer in the cells harvested whenever the medium was changed. RESULTS: After pretreatment of ovarian cancer cell lines (OVCAR-3 and SKOV-3) with 10 nM of staurosporine followed by treatment with 20 nM of paclitaxel, both OVCAR-3 and SKOV-3 cells were selectively arrested in G2M phase of cell cycle by paclitaxel and they resumed their proliferative cycle to some extents after the drugs were removed and cultured with fresh media. However. pretreatment with 10 nM of staurosporine protected normal circulating PBMC that had been induced to proliferate in vitro with phytohemagglutinin from paclitaxel. Staurosporine-induced arrest of PBMC in G0/G1 phase was reversible, and arrested cells tolerated 10 nM of paclitaxel in culture. CONCLUSION: OVCAR-3 and SKOV-3 cancer cells can be targeted specifically with paclitaxel, following staurosporine-mediated, selective and reversible G0/G1 arrest in PBMC.
Cell Cycle ; Cell Line ; Cytoprotection ; DNA ; Drug Therapy* ; Humans ; Ovarian Neoplasms ; Paclitaxel ; Staurosporine

Numerous studies have established a link between autophagy and aging; however, the relationship has not been clearly defined. Aging is a very complex process caused by the accumulation of various factors due to the gradual failure of cellular maintenance. Recent studies have shown that autophagy reduces the stress responses induced by starvation, reactive oxygen species, and the accumulation of intracellular proteins and organelles through cytoprotection, clearance of damaged mitochondria, and lysosomal degradation. Here, we summarize our current understanding of the relationship between autophagy and the aging process.
Aging* ; Autophagy* ; Caloric Restriction ; Cytoprotection ; Mitochondria ; Organelles ; Proteins ; Reactive Oxygen Species ; Starvation

Numerous studies have established a link between autophagy and aging; however, the relationship has not been clearly defined. Aging is a very complex process caused by the accumulation of various factors due to the gradual failure of cellular maintenance. Recent studies have shown that autophagy reduces the stress responses induced by starvation, reactive oxygen species, and the accumulation of intracellular proteins and organelles through cytoprotection, clearance of damaged mitochondria, and lysosomal degradation. Here, we summarize our current understanding of the relationship between autophagy and the aging process.
Aging* ; Autophagy* ; Caloric Restriction ; Cytoprotection ; Mitochondria ; Organelles ; Proteins ; Reactive Oxygen Species ; Starvation

PURPOSE: To evaluate the protective effect of creatine on the survival of retinal ganglion cells after serum deprivation. METHODS: RGC-5 cells were exposed to 5 mM creatine with serum-free media for 4 days. Cellular survival and mitochondrial respiratory activity were measured with MTT assay and resazurin assay, respectively. Degree of apoptosis was evaluated with vital staining using acridine orange/Hoechest 33342 and flow cytometric analysis using annexin/PI, respectively. RESULTS: Creatine increased cellular survival of RGC-5 cells significantly after serum deprivation. Additionally, creatine increased mitochondrial respiratory activity and inhibited apoptosis of RGC-5 cells. CONCLUSIONS: The energy precursor creatine increased survival of retinal ganglion cells after serum deprivation. Creatine could be relevant for the cytoprotection of retinal ganglion cells.
Apoptosis ; Creatine ; Culture Media, Serum-Free ; Cytoprotection ; Oxazines ; Retinal Ganglion Cells ; Xanthenes

OBJECTIVETo evaluate the performance of a new highly efficient and environment-friendly tissue cell fixatives for preserving the morphologies and properties of pleural and peritoneal effusions.

METHODSFifty-six specimens of tissue cells from pleural and peritoneal effusions were preserved using the new preservative or 95% ethanol. HE staining and Western blotting were employed to detect the morphologies and protein expression levels of CK, CEA and P53 of the cells after fixation.

RESULTSThe new preservative well preserved the morphologies of the cells from the pleural and peritoneal effusions, and the nuclei and cytoplasm were intact with little debris. The conventional preservative (95% ethanol) caused noticeable structural damage of the tissue cells, especially the cytoplasm where obvious debris were seen after fixation. CK, CEA and P53 protein expression levels in the cells were 91%, 86% and 88% after fixation with the new preservative, significantly higher than those (46%, 38% and 31%, respectively) in cells fixed with 95% ethanol (P<0.05).

CONCLUSIONThe new preservative is efficient and environment-friendly for preserving the morphologies as well as the proteins of tissue cells from pleural and peritoneal effusions well, demonstrating its potential in tissue cell fixation and preservation.

It has been an essential trend to understand and solve the difficult problems arising in the treatment process of burn with views of holistic theory. Recent researches have indicated that the driven factors and the termination signals of repair system engineering in treatment of burn are the unity of two opposite rather than two independent bodies with chronological order. Repair driven factors are germinated at the cost of systemic inflammatory response and even multiple organ damage. Inflammatory response is both a necessary procedure of burn repair and the pathological basis of multiple system dysfunction after burn. A comprehensive burn therapy nominated sequential cytoprotection (SCP) strategy has emerged in which the knowledge derived from basic research is translated to clinical practice stepwise, and it might play an important role in treatment of severe burn. Further multi-center randomized controlled clinical trials should be conducted in order to raise the level of SCP strategy in guideline of evidence-based medicine.
Burns ; therapy ; Cytoprotection ; Evidence-Based Medicine ; Humans ; Reconstructive Surgical Procedures ; Wound Healing

BACKGROUND AND OBJECTIVES: Heat shock protein (HSP) is an intracellular protein, expressed for cell protection under stressful condition. Expression of HSP was found in cochlear after noise stimulation. The vestibular end organs, especially saccule, have been known to be damaged by loud sound because of its anatomical and embryological vicinity with the cochlea. In this study, we observed the function of HSP72 that it is indeed expressed in rat saccule following noise stimulation. MATERIALS AND METHODS: Experimental animals were exposed to noisy environment at 100, 110 and 120 dB SPL for 90 minutes. Noise stimulation was transferred through a tone hook of behind-the-ear type hearing aid. Saccule was dissected and stained immunohistochemically and examined for expression of HSP under light microscope. RESULTS: Strong immunoreactivities were observed at type I and II hair cells of rat saccule which received noise of 120 dB for 90 minutes. Stronger imm-unoreactivities were found along the marginal areas of the saccule. CONCLUSION: These findings suggest that HSP72 are expressed in rat saccule following overthreshold noise and it could be presumed that HSP72 may have a protective role against noise trauma.
Animals ; Cochlea ; Cytoprotection ; Hair ; Hearing Aids ; Heat-Shock Proteins* ; Hot Temperature* ; Noise* ; Rats* ; Saccule and Utricle*

BACKGROUND AND OBJECTIVES: We investigated the ability of a brief heat shock on day one to provide delayed protection against lethal heat stress on day two in a rat-derived H9c2 cardiomyoblast cell line with reference to the role of heat shock protein 25/27, 70i and the p38MAPK signalling pathway. MATERIALS, METHODS AND RESULTS: Heat preconditioning(Heat P; 20min at 42+/-0.1degrees C) and adenosine(ADO) administered on day 1 protected against cell death under lethal heat challenge (75min at 42+/-0.1degrees C) on day 2 as measured by MTT test: ( % cell viability: Heat P: 79.9+/-3.23%, ADO: 71.9+/-4.10% vs. control: 52.7+/-1.65% respectively ). This protection was abolished by treatment with SB203580 or cytochalasine D prior to the protective stimulus on day 1( SB203580: 64.1+/-4.37%, cytochalasine D: 73.1+/-4.33% vs. Heat P ). Western blotting analysis indicated a significant accumulation of hsp70i in Heat P and SB203580-treated Heat P cells compared to control and adenosine-, SB203580-treated cells. Phosphorylation of hsp25 was significantly increased in Heat P cells compared to control cells. We also observed fragmentation of F-actin and formation of F-actin aggregates in cells exposed to lethal heat challenge. In contrast, the delayed cytoprotection preserved the F-actin bundles under lethal heat challenge. Hsp27-overexpressed, stable clones were more resistant to lethal heat shock when compared to control cells transfected with the vector alone. CONCLUSION: These data suggest an important role for p38MAPK/hsp25/27 pathway as a potential distal effector of heat-induced delayed protection.
Actins ; Blotting, Western ; Cell Death ; Cell Line ; Cell Survival ; Clone Cells ; Cytoprotection ; Cytoskeleton* ; Heat-Shock Proteins* ; Hot Temperature* ; Phosphorylation ; Shock