Cytotoxic Activity of Ethanolic Extract Aquilaria malaccensis Leaves Against MCF-7 Cells

@#Introduction: Aquilaria malaccensis, also known as “Pokok Karas” in Malaysia, is widely used in Southeast Asian countries for the treatment of joint pain, diarrhoea and inflammatory diseases, and has shown beneficial effects as an anticancer agent. The aim of this study was to investigate the effect of ethanol leaf extracts of A. malaccensis on MCF-7 cells. Methods: MTT-based cytotoxic and antiproliferative assay was used to determine the outcome of ethanolic extract toward MCF-7 cells. The mode of cell death was determined by the AO/PI double staining assay and the depolarisation of the mitochondria membrane potential. Results: IC50 value of the extract against MCF-7 cells treated for 72 hours was 4.1 ± 2.08 µg/mL, while the IC50 value for doxorubicin was 2.92 ± 0.12 µg/mL. The extract showed a lower cytotoxic effect against the NIH/3T3 cells and inhibited the growth of MCF-7 cells in a dose dependent manner. AO/PI double stain showed that the ethanolic extract of A. malaccensis leaves induced MCF-7 cells into apoptotic cell death. The present study showed that the ethanolic extract of A. malaccensis induced apoptosis through mitochondrial pathway as indicated by its ability to take up JC-1. Conclusion: The study found that ethanolic extract obtained from A. malaccensis leaves is cytotoxic on MCF-7 cells, resulting to apoptotic cell death of the cells.

The protective effect of water-soluble proteins from selenium-enriched Bifidobacterium longum on lipopolysaccharide-induced injury in rat IEC6 cells / 中国药理学通报

To investigate the protective effect of selenium-enriched Bifidobacterium longum preparation on rat IEC6 cells via the application of lipopolysaccharide (LPS)-induced IEC6 cell injury model. Methods The experiment was divided into five groups; negative control group treated with phosphoric acid equilibrium salt solution, model group treated with LPS, and low, medium, high concentration group in which IEC6 cells were treated with LPS plus 10, 30, 100 mg • L"1 water-soluble proteins from selenium-enriched Bifidobacterium longum respectively. After treatment with LPS for 24 hours, IEC6 cell viability, apoptosis, mitochondria membrane potential, and the expression of zonula occludens 1 (ZO-1) and Occludin were detected. Results LPS induced rat intestinal epithelial cell damage, such as the decrease of cell viability, the increase of cell apoptosis, the collapse of mitochondria membrane potential, and the decrease of cell tight junction protein expression. Water-soluble proteins from selenium-enriched Bifidobacterium longum inhibited LPS-induced IEC6 cell damage, decreased cell apoptosis and increased cell tight junction between cells. Conclusion Water-soluble proteins from selenium-enriched Bifidobacterium longum have protective effect on LPS-induced rat intestinal epithelial cell injury.

Regulation of Wnt5a in apoptosis of human lung adenocarcinoma A549 cells and its mechanism / 吉林大学学报(医学版)

Objective: To investigate the regulation effect of Wnt5a on the apoptosis of lung adenocarcinoma A549 cells, and to clarify its mechanism. Methods, The human lung adenocarcinoma cells were selected. The A549 cells treated with Wnt5a were used as treatment group, and the A549 cells treated with C culture solution were used as control group. The apoptotic body induced by Wnt5a was assessed with TUNEL assay; the apoptotic rates of the A549 cells in various groups were detected by Annexin V-FITC/PI double staining; the reactive oxygen species (ROS) levels in the A549 cells in various groups were determined with DCFH-DA fluorescence probe, and the mitochondria membrane potential was assessed with JC-1 staining method. Western blotting was used to analyze the expression levels of apoptosis-related proteins in the A549 cells in various groups. Results; Compared with control group, the apoptotic rates of the A549 cells in treatment group 12, 24, and 48 h after treatment were significantly increased (P<0.01); the ROS levels were increased (P<0.05); the mitochondria membrane potentials were decreased (P<0.05), the expressing amount of BAX was up-regulated and the expression amount of AIF was down-regulated. Conclusion: Wnt5a has regulation on the apoptosis of human lung adenocarcinoma cells and can promote the apoptosis of A549 cells through mitochondrial pathway.

Regulation of Wnt5a in apoptosis of human lung adenocarcinoma A549 cells and its mechanism / 吉林大学学报(医学版)

Objective:To investigate the regulation effect of Wnt5a on the apoptosis of lung adenocarcinoma A549 cells,and to clarify its mechanism.Methods:The human lung adenocarcinoma cells were selected.The A549 cells treated with Wnt5a were used as treatment group,and the A549 cells treated with C culture solution were used as control group.The apoptotic body induced by Wnt5a was assessed with TUNEL assay;the apoptotic rates of the A549 cells in various groups were detected by Annexin V-FITC/PI double staining;the reactive oxygen species (ROS)levels in the A549 cells in various groups were determined with DCFH-DA fluorescence probe,and the mitochondria membrane potential was assessed with JC-1 staining method.Western blotting was used to analyze the expression levels of apoptosis-related proteins in the A549 cells in various groups.Results:Compared with control group,the apoptotic rates of the A549 cells in treatment group 12,24,and 48 h after treatment were significantly increased(P<0.01);the ROS levels were increased(P<0.05);the mitochondria membrane potentials were decreased(P<0.05),the expressing amount of BAX was up-regulated and the expression amount of AIF was down-regulated.Conclusion:Wnt5a has regulation on the apoptosis of human lung adenocarcinoma cells and can promote the apoptosis of A549 cells through mitochondrial pathway.

The changes of mitochondrial membrane potential detected by confocal microscopy in spermatocyte induced by bisphenol A / 中国医学装备

Objective:To detect changes in mitochondrial membrane potential after sperm mother cell damage induced by bisphenol A by laser scanning confocal microscopy(LSCM) and underline its potential action mechanism.Methods: The cultured spermatogenic cells were divided into 3 groups, respectively. Then 0, 10μmol/L, 100μmol/L of Bisphenol A(BPA) were added into the culture, after 3 hours culture, fluorescence probe JC-1 was used to lable the three groups. The fluorescence intensity of JC-1 in mitochondrial was then detected by LSCM. LSCM software was used to analyze the fluorescence intensity. Change of the mitochondrial membrane potential was represented by the change of fluorescence colors (relative proportion of red and green fluorescence is commonly used to measure mitochondrial depolarization ratio). Results:The ratio between the red and green fluorescence in the control group, low dose group and high dose group had significant difference. Intracellular mitochondrial membrane potential of Bisphenol A treatment group was lower than that of the control group, while mitochondrial membrane potential of high dose group was lower than that of the low dose group. Conclusion: Bisphenol A could damage spermatocytes, and as the dose increased, the more serious the injury. The method could real-time monitor with high sensitivity the change of intracellular mitochondrial membrane potential.

Protective effect of trigonelline on rat cardiomyocytes during hypoxia and reoxygenation / 重庆医学

Objective To investigate the protective effects of trigonelline in neonatal rat cardiomyocytes during hypoxia /reoxygenation and its mechanism .Methods The neonatal rat cardiomyocytes was randomly divided into the normal control group , the hypoxia/reoxygenation group and the trigonelline group .Flow cytometry was used to determine the apoptosis and mitochondrial membrane potential .The levels of SOD and MDA were measured in different groups .Western blot method was used to measure the procaspase‐9 ,cleaved caspase‐9 ,procaspase‐3 and cleaved caspase‐3 protein level .Results Trigonelline could inhibit apoptosis (P<0 .05) ,enhanced activity of SOD (P<0 .05) ,reduce production of MDA (P<0 .05) ,stabilize the mitochondrial membrane potential (P<0 .05) and activate caspase‐9 and caspase‐3 in neonatal rat cardiomyocytes during hypoxia/reoxygenation .Conclusion Trigo‐nelline could protect myocardial cells from injury caused by hypoxia and reoxygenation ,and the mechanism may be associated with anti‐lipid peroxidation and stabilizing mitochondria membrane potential .

Anticancer effect of xanthono-pyridine derivative xp-16 on human gastric carcinoma MGC-803 cells / 中国药学杂志

OBJECTIVE: To investigate aticancer effect and potential mechanism of a new xanthono-pyridine derivative N, N'-(7-oxo-7H-chromenoquinoline-5, 9-diyl)-bis(2-(pyrrolidin-1-yl)acetamide) (XP-16) on human gastric carcinoma cell line MGC-803.

Fucodiphlorethol G Purified from Ecklonia cava Suppresses Ultraviolet B Radiation-Induced Oxidative Stress and Cellular Damage

Fucodiphlorethol G (6'-[2,4-dihydroxy-6-(2,4,6-trihydroxyphenoxy)phenoxy]biphenyl-2,2',4,4',6-pentol) is a compound purified from Ecklonia cava, a brown alga that is widely distributed offshore of Jeju Island. This study investigated the protective effects of fucodiphlorethol G against oxidative damage-mediated apoptosis induced by ultraviolet B (UVB) irradiation. Fucodiphlorethol G attenuated the generation of 2, 2-diphenyl-1-picrylhydrazyl radicals and intracellular reactive oxygen species in response to UVB irradiation. Fucodiphlorethol G suppressed the inhibition of human keratinocyte growth by UVB irradiation. Additionally, the wavelength of light absorbed by fucodiphlorethol G was close to the UVB spectrum. Fucodiphlorethol G reduced UVB radiation-induced 8-isoprostane generation and DNA fragmentation in human keratinocytes. Moreover, fucodiphlorethol G reduced UVB radiation-induced loss of mitochondrial membrane potential, generation of apoptotic cells, and active caspase-9 expression. Taken together, fucodiphlorethol G protected human keratinocytes against UVB radiation-induced cell damage and apoptosis by absorbing UVB radiation and scavenging reactive oxygen species.

XP-16, a new xanthono-pyridine derivative, induces apoptosis in human lung carcinoma A549 cells / 中国药理学通报

Aim To investigate the anticancer effect of a new xanthono-pyridine derivative N, N '-( 7-oxo-7H-chromeno[3,2-h] quinoline-5,9-diyl)-bis(2-( pyrroli-din-1-yl)acetamide) (XP-16) on human lung carcino-ma cell line A549 and the potential mechanism. Meth-ods Antiproliferative effect of XP-16 on A549 cells was evaluated by MTT assay, morphological examina-tion and colonial assay. Apoptosis detection was car-ried out using Hoechst 33258 and PI double-dyeing method. Intracellular Ca2+ concentration ( [ Ca2+] i ) and mitochondria membrane potential were detected by fluorospectrophotometer. A549 cells treated with XP-16 were collected for Bad and metallothionein 1 A ( MT-1 A ) transcript analysis by real-time reverse tran-scriptase-polymerase chain reaction ( qRT-PCR) . Re-sults XP-16 inhibited A549 cell proliferation in dose-and time-dependent manner. Typical apoptotic mor- phology such as chromatin aggregation and nuclear fragmentation was observed in A549 cells treated with XP-16 for 24 h, and the apoptosis was showed in a dose-dependent manner. After treated with XP-16, [ Ca2+] i and mitochondria membrane potential of A549 cells were decreased, and relative mRNA level of Bad and MT-1A was up-regulated. Conclusions XP-16 has anticancer effect on A549 cells through apoptosis, which might be associated with decreasing intracellular Ca2+ concentration and mitochondria membrane poten-tial. Up-regulation of MT-1A expression might be the result of decreased [ Ca2+] i .

Glutamine attenuates the production of nitric oxide production and the reduction of mitochondrial membrane potential of cultured rat hepatocytes induced by IL-1β / 中华急诊医学杂志

Objective To investigate the effects of different concentrations of glutnine(Gln) on the procfion of nitric oxide (NO)and reduction in mitochondrial membrane potential in rat heputocytesactivated in by interleukin(IL)-1β.Method The primary cultured rat hepatocytes with high-purity Wfffe isolated from 6-week-old male Sprague-Dawley raL, by flirt in situ circulatory collagenase peffusion method.After incubation for hours.hepatocytes were stimulated by saline,or IL-l(1 nmol/L)or IL-I(1 mnol/L)combined with gIne in concentrations rangiIfrom 2 to 10 mmol/L.The culture muln and hepatocytes were collected at 24 hours after stimulation.The concentrations of alanine aminotransferase(ALT)and NO in the lnllln were detected by biochemical methods.The mitochondrial membrane potential of the hepatocytes was detected with flow cytometry after incubated with fluorescem probe JC-1.Statistic package ofSPSS 11.5 was used for the data analysand significant differences between mean8 were evaluated byQ、4k analysis.Results The average concentration ofand NOinthe culturemedium afterIL-Istimulationwas 38.2U/L and 72.7tmaol/L,respectively,whichwere sis,cantly big,herthanthose ofcontrol group(7.4 U/L and41.7nol/L,respectively,P<0.01).1hemitochondrial membrane potential ofhepatocytes in IL-lgroup was much lowerthan that in control group(30.O%vs.62.8%.P<0.01).Gin inhibited NO production induced byⅡ,lp,Jeleasing and reduction in mitochondrial II brahetential ofcuhured rat hepatocytes in a dose-dependent HmIll.Conclusions Ghtamine,the most abundant free amino acid in the body,call attenuate hepatocye injury mechateel in vitro by pro-inflammatory cytokine mediated.nlis protective effectmay be associatedwiththeinhibition of NO production and thereby amelioration of mitochondrialfunction.

MITOCHONDRIAL CHANGES AND THEIR REGULATION INMURINE APOPTOTIC PERITONEAL MACROPHAGE / 解剖学报

Objective To study the changes in activity of NADPH oxidase, the effects of signal molecules on membrane potential and ROS production of mitochondria in apoptotic murine peritoneal macrophages. Methods Laser scanning confocal microscopy, flow cytometry and fluorescence labeling were used. Results 1 The macrophages treated with dexamethasone developed apoptosis quickly and presented concomitant apoptotic changes. 2 Mitochondria membrane depolarized quickly, the activity of NADPH oxidase declined sharply, and ROS production decreased rapidly. The erasers of ROS promoted macrophage apoptosis. 3 PKC favored, and cAMP inhibited the macrophage apoptosis and the rapid drop in ROS and mitochondrial membrane depolarization. cGMP and TPK which slightly inhibited macrophage apoptosis, had no effects on ROS. Conclusion 1 The activity of NADPH oxidase declined sharply, hence the ROS decreased rapidly, which promoted apoptosis in macrophages treated with dexamethasone. 2 The signal molecules affected apoptosis by modulating ROS decline and mitochondria depolarization. The results suggested that, mitochondria variations, especially the variations of ROS and membrane potential, mainly affected macrophage apoptosis.;