Objective To investigate the effect of Zingiber corallinum oil(ZCO)on apoptosis and proliferation of cervical carcinoma cell line HeLa. Methods HeLa cells were treated with different concentrations of ZCO(5-80 mg·L-1)in vitro. Cytotoxicity rate was determined by CCK-8 assay. The morphological changes was observed using inverted microscope after AO/EB staining. Caspase-3 activities were measured with a colorimetric method. Protein level of hsp-70 were detected by Western blotting. Cell cycle and apoptosis were analyzed by flow cytometer (FCM).Results ZCO exhibited effect of proliferation inhibition and apoptosis-inducing on the growth of HeLa cells in a dose-dependent manner. Caspase-3 activities increased in a dose-dependent manner while the expression of hsp-70 decreased. Cell cycle was arrested in G2/M phase. Conclusion ZCO exhibites a marked effect of proliferation inhibition and apoptosis-inducing on HeLa cells. The mechanism of ZCO might be activating the key enzyme in apoptotic pathway, so that the expression of hsp-70 is down-regulated, and cell cycle is arrested in G2/M phase.
Fig.1
Morphology of four groups of HeLa cells (A0/EB staining,×200) A. normol control group;B. 5 mg·L-1 ZCO group;C. 20 mg·L-1 ZCO group;D. 80 mg·L-1 ZCO group
表2
Tab.2
表2
表2
4组HeLa细胞 48 h caspase-3酶活性比较
Tab.2
Comparison of caspase-3 activity among four groups of HeLa cells at 48 h x¯±s,n=6
组别
A值
酶活性
正常对照组
0.81±0.01
-
5 mg·L-1 ZCO组
0.72±0.00
1.05±0.12*1
20 mg·L-1 ZCO组
0.15±0.01
1.91±0.03*1
80 mg·L-1 ZCO组
0.07±0.00
3.31±0.01*1
Compared with normal control group , *1P<0.05
与正常对照组比较,*1P<0.05
表2
4组HeLa细胞 48 h caspase-3酶活性比较
Tab.2
Comparison of caspase-3 activity among four groups of HeLa cells at 48 h x¯±s,n=6
ANTTILAA, PUKKALAE, SODERMANB, et al.Effect of organised screening on cervical cancer incidence and mortality in Finland, 1963-1995: recent increase in cervical cancer incidence.[J]. ,1999,83(1):59-65.
A nation-wide screening programme for cervical cancer started in Finland gradually from 1963 onwards. By the beginning of the 1990s, there had been a decrease of 80% both in the age-adjusted incidence of and mortality from cervical cancer. To describe the recent patterns in cervical cancer incidence and mortality and evaluate their differentials in relation with the organised screening activities, we have updated the material on the cervical cancer incidence and mortality as well as mass-screening activities up to the year 1995. Based on the files of the Finnish Cancer Registry, there is a striking increase of about 60% in the incidence of cervical cancer during the last 4 years of the study period among women below 55 years of age. The mortality rates are still decreasing. There is no overall decrease over recent years in the coverage of the programme invitations or smears taken. Incidence of invasive cancer and of moderate and severe dysplasia as detected in mass screening have increased. As to the interpretation, changes in the risk factors, such as in sexual behaviour and smoking habits, over the decades might partly explain increasing trends in cervical cancer incidence. As the change in incidence was relatively abrupt, inadequacies or changes in the effectiveness in the screening programme, particularly among young women, may also have contributed. Expanding the coverage of and attendance in the pap-screening programme among women in young target ages would still be effective. Increasing emphasis on quality assessment in screening is also needed.
CHUNG HH, JANG MJ, JUNG KW, et al.Cervical cancer incidence and survival in Korea: 1993-2002[J]. ,2006,16(5):1833-1838.
Abstract.69 Chung HH, Hwang SY, Jung KW, Won YJ, Shin HR, Kim JW, Lee HP (On behalf of the members for Gynecologic Oncology Committee of Korean Society of Obstetrics and Gynecology). Ovarian cancer incidence and survival in Korea: 1993–2002. 2007;595–600.
YANGZ, LUOS, PENGQ, et al.GC-MS Analysis of the essential oil of coral ginger (Zingiber corallinum Hance) rhizome obtained by supercritical fluid extraction and steam distillation extraction[J]. ,2009,69(7/8):785-790.
The chemical compositions of volatile oils extracted from Zingiber corallinum Hance were obtained by steam distillation extraction (SDE) and supercritical fluid extraction (SFE) followed by gas chromatography-mass spectrometry analysis. The effects of pressure, temperature, modifier volume, and extraction time on the SFE of Z. corallinum oil were investigated, and optimization of the conditions were: extraction temperature, 3002°C; dynamic extraction time, 4002min; pressure, 1002Mpa; and modifier volume, 5002mL. 29 compounds from SFE extracts were identified. The main components were sabinene (16.19%), terpinen-4-ol (16.26%), β -sesquiphellandrene (3.26%), triquinacene 1, 4-bis (methoxy) (22.01%), triquinacene 1,4,7-tris (methoxy) (3.66%), 1-[4-(1-methyl-2-propenyl) phenyl] ethanone (9.87%) and ferulic acid methyl ester (16.88%) accounting for 88.13% of all compounds. SFE extracts were found to be markedly different from the corresponding SDE extracts. 22 compounds from SDE extracts were identified. The major components, accounting for 92.48% of SDE extracts, were sabinene (53.38%), α -terpinene (3.23%), γ -terpinen (2.16%), terpinen-4-ol (22.66%), β -sesquiphellandrene (1.41%) and triquinacene, 1, 4-bis (methoxy) (9.64%).
LUO SQ, PENG QC, YANG XO, et al.Volatiles and inhibitory phytopathogens fungi activities of essential oil extracted from Zingiber corallinum Hance[J]. ,2013,40(17):84-86.
To investigate antimicrobial activities of essential oil of Zingiber corallinum Hance against phytopathogenic fungi, the essential oil hydrodistillated was analyzed by gas chromatography- mass spectrometry(GC-MS). Using microbial growth inhibition assays in vitro, the oil was tested for fungitoxic effects against 4 phytopathogenic fungi, which were Rhizoctonia solani K眉hn, Fusarium gram inearum, Fusarium nivale, Pyricularia oryzae. Eighteen different components representing 91.87% of the compounds in the oil were identified. Sabinene(54.07%), 1-Terpinen-4-ol(23.74%), Triquinacene,1,4-bis(methoxy)-(6.47%) and 纬-Terpinen(4.60%) were found to be predominant components. The oil exhibited considerable antifungal activity against tested fungi, median effectively lethal concentration(EC50) of the oil against 4 phytopathogenic fungi were 3.64, 11.20, 11.33, 14.19 mg/L, respectively, and it was most significantly toxic against Rhizoctonia solani K眉hn. The research results provided theoretical basis for the development of environment friendly pesticide and comprehensive application of Zingiber corallinum Hance.
DE-SHUN YU, FEI-FEI YE, YANG XQ. Antioxidant activity of essential oil from Zingiber corallinum Hance[J]. ,2011,32(17):164-167.
A natural plant essential oil was extracted from Zingiber corallinum Hance as a common herb for Miao ethnic minority in Guizhou by supercritical carbon dioxide extraction.The antioxidant activity of the essential oil was evaluated by comparing changes in the peroxide value(POV) of concentrated fish oil during storage at 60-65 鈩 in the respective presence of the essential oil,vitamin E(VE) and butylated hydroxytoluene(BHT).Forty-four compounds were identified in the essential oil by GC-MS,including 13 monoterpenoids(25.12%),16 oxygen-containing monoterpenoids(26.43%),9 sesquiterpenes(8.82%) and 6 oxygen-containing sesquiterpenes,benzene derivatives or others(39.63%).The essential oil as a natural antioxidant was superior to VE and BHT.The fish oil can be protected effectively by adding 0.3% essential oil.
YANGC, ZHOU LL, WANG HY, et al.The inhibitory effect of Zingiber corallinum Hance essential oil on drug-resistant bacteria and evaluation of its acute toxicity[J]. , 2011,17(5):139-146.
BACKGROUND: The excessive and irregular use of antibiotics could result in the generation and diffusion of drug-resistant bacteria. The aim of this study was to investigate the inhibitory effect of Zingiber corallinum Hance essential oil (ZCHO) on drug-resistant bacteria, especially on drug-resistant Acinetobacter baumannii. MATERIAL/METHODS: Susceptibility testing was used to evaluate the effect of ZCHO on growth inhibition of drug-resistant bacteria by paper disk method. Mice orally administered with ZCHO were used to observe acute toxicity and to determine median lethal dose (LD6368) of ZCHO. Broth dilution method was used to determine minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of ZCHO on drug-resistant Acinetobacter baumannii. RESULTS: ZCHO exhibited an obvious inhibitory effect not only on gram-negative drug-resistant bacteria including Escherichia coli, Klebsiella pneumoniae, Enterobacter cloacae and Acinetobacter baumannii, but also on gram-positive drug-resistant bacteria including Staphylococcus aureus, Staphylococcus epidermidis and Staphylococcus haemolyticus. The ZCHO containing 79% terpinen-4-ol revealed better bacteriostatic effect than ZCHO with 34% terpinen-4-ol. The LD6368 of ZCHO was 1790.427 mg/kg. The MIC and MBC of ZCHO on drug-resistant Acinetobacter baumannii were 1457.81 mg/L. CONCLUSIONS: ZCHO has obvious bacteriostasis and bactericidal effects, especially against drug-resistant Acinetobacter baumannii. Therefore, ZCHO is a promising natural bioactive component with antibacterial effect and satisfactory safety due to its low toxicity.
LOOVEREN MV, GOOSSENSH, GROUP T A S. Antimicrobial resistance of Acinetobacter spp. in Europe[J]. ,2004,10(8):684-704.
Bacteria of the genus Acinetobacter are ubiquitous in nature. These organisms were invariably susceptible to many antibiotics in the 1970s. Since that time, acinetobacters have emerged as multiresistant opportunistic nosocomial pathogens. The taxonomy of the genus Acinetobacter underwent extensive revision in the mid-1980s, and at least 32 named and unnamed species have now been described. Of these, Acinetobacter baumannii and the closely related unnamed genomic species 3 and 13 sensu Tjernberg and Ursing (13TU) are the most relevant clinically. Multiresistant strains of these species causing bacteraemia, pneumonia, meningitis, urinary tract infections and surgical wound infections have been isolated from hospitalised patients worldwide. This review provides an overview of the antimicrobial susceptibilities of Acinetobacter spp. in Europe, as well as the main mechanisms of antimicrobial resistance, and summarises the remaining treatment options for multiresistant Acinetobacter infections.
HWANG TS, HAN HS, CHOI HK, et al.Differential, stage-dependent expression of Hsp70, Hsp110 and Bcl-2 in colorectal cancer[J]. ,2003,18(6):690-700.
The presence of hypoxic cells in solid tumors has been suggested to contribute to the malignant progression of various tumors. Recently, we reported an activation of heat shock transcription factor (HSF) and expression of heat shock proteins (Hsp) in murine tumor cells by hypoxia.To search for a possible role of Hsp in the malignant progression of human tumors, we analyzed the expression profiles of Hsp family proteins in weakly and highly metastatic human colorectal cancer (CRC) cell lines. We also analyzed the expression profiles of Bcl-2 family proteins because the altered expression of these proteins has been demonstrated in various solid tumors.In the present paper we showed among various Hsp and Bcl-2 family proteins that the expression of Hsp70 and Hsp110 was elevated in highly metastatic CX-1 and HT-29 cells, while the expression of Bcl-2 was elevated in weakly metastatic MIP-101 and Clone A cells. Subsequent immunohistochemical analysis of 81 primary human CRC tissues demonstrated that the expression of Hsp70 and Hsp110 was highly correlated with the advanced clinical stages and positive lymph node involvement. The expression of Bcl-2, in contrast, was correlated to the early clinical stage and negative lymphovascular invasion.Taken together, our study demonstrated for the first time a differential, stage-dependent expression of Hsp70, Hsp110 and Bcl-2 in CRC. We suggest that the molecular mechanisms underlying the differential expression of Hsp and Bcl-2 family members deserves a more rigorous future study, the results of which might offer novel modes of rationale and strategy to predict and manipulate the malignant progression of colorectal cancers.
CAOX, WANG AH, JIAO RZ, et al.Surfactin induces apoptosis and G2/M arrest in human breast cancer MCF-7 cells through cell cycle factor regulation[J]. ,2009,55(3):163-171.
Surfactin, purified from Bacillus subtilis natto TK-1, inhibited proliferation of human breast cancer MCF-7 cells in a dose- and time-dependent manner, with IC 50 at 24, 48, and 72聽h of 82.6, 27.3, and 14.8聽M, respectively. Surfactin-induced cell death was considered to be apoptotic by observing the typical apoptotic morphological change by acridine orange/ethidium bromide staining and Transferase-mediated dUTP Nick End-labeling assay. [Ca 2+ ]i measurement revealed that surfactin induced a sustained increase in concentration of intracellular [Ca 2+ ]i. Flow cytometric analysis also demonstrated that surfactin caused time-dependent apoptosis of MCF-7 cells through cell arrest at G 2 /M phase. Western blot revealed that surfactin induced accumulation of the tumor suppressor p53 and cyclin kinase inhibitor p21 waf1/cip1 , and inhibited the activity of the G 2 -specific kinase, cyclin B1/p34 cdc2 . Based on our findings, surfactin inhibited proliferation in MCF-7 cells by inducing apoptosis and the elevation of [Ca 2+ ]i may play an important role in the apoptosis. The mechanism which surfactin caused G 2 /M arrest seems to be through cell cycle factor regulation.
DEACIUC IV, FORTUNATOF, D'SOUZA N B, et al. Modulation of caspase-3 activity and Fas ligand mRNA expression in rat liver cells in vivo by alcohol and lipopolysaccharide[J]. ,1999,23(2):349-356.
The purpose of this study was to determine if exacerbation of apoptosis precedes liver injury during chronic exposure of rats to alcohol. After 7 weeks of feeding an alcohol- or dextrin-containing liquid diet, the animals were treated with gram-negative bacterial lipopolysaccharide (1 mg x kg(-1) body weight, intravenously) or sterile saline and sacrificed 3 hr after the treatment. Alanine:2-oxoglutarate aminotransferase (ALT) and lactate:NAD oxidoreductase [lactate dehydrogenase (LDH)] were measured in plasma. The caudate lobe of the liver was resected for histology, while the rest of the organ was perfused with collagenase to isolate hepatocytes, Kupffer cells (KCs), and sinusoidal endothelial cells (SECs) by centrifugal elutriation. Hepatocyte mitochondria were isolated by differential centrifugation of the cell homogenate. Reduced and oxidized glutathione (GSH and GSSG) in isolated hepatocytes and hepatocyte mitochondria, and malondialdehyde in hepatocytes were assayed. Caspase-3 activity and Fas ligand mRNA expression were determined in hepatocytes, KCs, and SECs. Plasma ALT and LDH activity, liver histology, GSH, GSSG and their ratio, and malondialdehyde content were not affected by alcohol treatment Caspase-3 activity was significantly increased in alcohol-treated rats in all three cell types, with the lowest response observed in hepatocytes and the highest in KCs. Fas ligand mRNA expression, which had the highest level in SECs, followed by KCs and hepatocytes, was not affected by alcohol administration. Lipopolysaccharide had the following effects: an increase in ALT in both pair- and alcohol-fed rats, and LDH only in alcohol-fed rats, a decrease in GSH + GSSG levels in both mitochondria and hepatocytes, an elevation of malondialdehyde content in hepatocytes, a raise in caspase-3 activity in all groups and cell types, and an augmentation of Fas ligand expression in hepatocytes and KCs, but not in SECs. These data suggest that, during chronic alcohol consumption, an exacerbated apoptosis precedes alcohol-induced liver injury.
LU HF, SUE CC, YU CS, et al.Diallyl disulfide (DADS) induced apoptosis undergo caspase-3 activity in human bladder cancer T24 cells[J]. ,2004,42(10):1543-1552.
Diallyl disulfide (DADS), one of the major components of garlic (Allium sativum), is well known to have chemopreventative activity against human cancer such as colon, lung and skin. But the exact mechanism of the action is still unclear. In this study, we investigated how DADS––induced cell cycle arrest and apoptosis in T24 human bladder cancer cells in vitro. Apoptosis induction, cell viability, cell cycle arrest, caspases-3, -9 activity and gene expression were measured to determine their variation by flow cytometric assay, western blot, and determination of caspase-3 activity, PCR and cDNA microarray. There are significant differences in cell death (decreased viable cells then increased the amounts of apoptosis) of T24 cells that were detected between DADS (5–75 μM) treated and untreated groups. A significant increase was found in apoptosis induction when cells were treated with DADS (50 μM) compared to without DADS treated groups. DADS also promoted caspase-3 activity after exposure for 1, 3, 6, 12, and 24 h, which led to induce apoptosis. DADS also increased the product of intracellular hydrogen peroxide. Furthermore, the DADS-induced apoptosis on T24 cells was blocked by the broad-spectrum caspase inhibitor, z-VAD-fmk and antioxidant (catalase). DADS also increased cyclin E and decreased CDK2 gene expression which may lead to the G2/M arrest of T24 cells.
IRAJ JA, MOJTABAS, FATEMEHV, et al.Evaluation of insulin and ascorbic acid effects on expression of Bcl-2 family proteins and caspase-3 activity in hippocampus of STZ-induced diabetic rats[J]. ,2009,29(1):133-140.
We assessed the expression of Bcl-2 family members at both mRNA and protein levels as well as the Caspase-3 activity, in order to investigate the occurrence of apoptosis in hippocampus of STZ-induced diabetic rats. We selected twenty-four Wistar rats; half of them were made diabetic by intraperitoneal injection of a single 60 mg/kg dose of streptozotocin (STZ, IP), while the others received normal saline and served as controls. The expressions of Bcl-2, Bcl-x(L), and Bax mRNA and proteins were measured using RT-PCR and western blotting, respectively. Caspases-3 activity was determined by using the Caspase-3/CPP32 Fluorometric Assay Kit. The result showed that mRNA and protein levels of Bcl-2 and Bcl-x(L) were lower in hippocampus of diabetic group than that of the control group, whereas expressions of Bax in hippocampus of diabetic rats were higher than that of controls at both mRNA and protein levels (P < .01). Hyperglycemia was found to raise 6.9-fold hippocampal caspase-3 activity in diabetic group compared with control group (P < .001). Therefore, the induction of diabetes is associated with increased ratios of Bax/Bcl-2, Bax/Bcl-x(L), and increased caspase-3 activity in hippocampus which shows that apoptosis is favored in hippocampal region.
MOSSER DD, CARON AW, BOURGETL, et al.The chaperone function of hsp70 is required for protection against stress-induced apoptosis[J]. ,2000,20(19):7146-7159.
Cellular stress can trigger a process of self-destruction known as . Cells can also respond to stress by adaptive changes that increase their ability to tolerate normally lethal conditions. Expression of the major heat-inducible protein protects cells from heat-induced . has been reported to act in some situations upstream or downstream of , and its protective effects have been said to be either dependent on or independent of its ability to inhibit activation. Purified has been shown to block procaspase processing in vitro but is unable to inhibit the activity of active caspase 3. Since some aspects of function can occur in the absence of its chaperone activity, we examined whether lacking its ATPase domain or the C-terminal EEVD sequence that is essential for was required for the prevention of . We generated stable cell lines with -regulated expression of , , and chaperone-defective mutants lacking the ATPase domain or the C-terminal EEVD sequence or containing AAAA in place of EEVD. Overexpression of or protected cells from heat shock-induced by preventing the processing of procaspases 9 and 3. This required the chaperone function of since mutant proteins did not prevent procaspase processing or provide protection from . activation was inhibited by both and and by each of the domain mutant proteins. The chaperoning activity of is therefore not required for inhibition of activation, and inhibition was not sufficient for the prevention of . Release of from mitochondria was inhibited in cells expressing full-length but not in cells expressing the protein with ATPase deleted. Together with the recently identified ability of to inhibit -mediated procaspase 9 processing in vitro, these data demonstrate that can affect the apoptotic pathway at the levels of both release and initiator and that the chaperone function of is required for these effects.
ELDER RT, BENKOZ, ZHAOY.HIV-1 VPR modulates cell cycle G2/M transition through an alternative cellular mechanism other than the classic mitotic checkpoints.[J]. ,2002,7(2):d349-d357.
Abstract HIV-1 Vpr induces cell cycle G2/M arrest in both human and fission yeast cells, suggesting a highly conserved activity of this viral protein. In this review, we summarize the current understanding of Vpr-induced G2 arrest based on studies from both mammalian cells and the fission yeast (Schizosaccharomyces pombe) model system. Fission yeast has proven to be an excellent model system to investigate cell cycle G2/M control of eukaryotic cells. Similarly, fission yeast has also been instrumental in defining the molecular mechanism underlying the G2 arrest induced by Vpr. We have compared the classic DNA-damage and DNA-replication checkpoint controls of the cell cycle G2/M transition to the G2 arrest conferred by Vpr. Based on the current findings, we hypothesize that Vpr induces cell cycle G2 arrest through an alternative novel cellular pathway(s) rather than through the classic mitotic checkpoint controls. A number of cellular proteins which may be involved in this new cellular pathway(s) have been identified and are discussed.
GC-MS Analysis of the essential oil of coral ginger (Zingiber corallinum Hance) rhizome obtained by supercritical fluid extraction and steam distillation extraction
Evaluation of insulin and ascorbic acid effects on expression of Bcl-2 family proteins and caspase-3 activity in hippocampus of STZ-induced diabetic rats