Gene Expression Levels Of Caspase-3 Research Articles

Hyperbaric oxygen mitigates KIM-1 and inflammatory cytokine levels in kidney transplantation

The aim of this study was to investigate the effect of hyperbaric oxygen (HBO2) administration applied during cold ischemic time to organs removed from donors before kidney transplantation. A total of 24 rats were divided into three groups: Group 1 was the control group, Group 2 received 60 minutes of HBO2 at 2.5 atmospheres absolute, and Group 3 received 120 minutes of 2.5 ATA HBO2. The renal artery was entered with a polyethylene catheter and perfused with a standard organ preservation solution. Falcon tubes containing organs obtained from rats in Groups 2 and 3 were placed in a box supported by ice blocks. The temperature was kept constant at 4 °C and the box was placed in a pressure tank with 2.5 ATA HBO2. HBO2 was applied for 60 and 120 minutes, respectively. Organ samples were harvested at the end of 24 hours for histopathological evaluation, immunohistochemical analysis of TNF-α and IL-18, TUNEL analysis for apoptosis, and gene expression levels of kidney injury molecule-1 (KIM-1) and caspase-3. In histopathological examinations, hematoxylin and eosin staining was performed and samples were evaluated for tubular necrosis and vacuolization criteria. Group 2 and Group 3 had significant decreases compared to Group 1 in this regard. Immunohistochemical staining was performed for TNF-α, IL-18, and apoptosis levels; significant decreases were found in Groups 2 and 3. There were significant decreases in Groups 2 and 3 for KIM-1 and caspase-3 gene expression levels compared to Group 1, as well. Thus, it was demonstrated that during the cold ischemic period before kidney transplantation, HBO2 administration to organs removed from donors can reduce apoptotic cell numbers, inflammatory cytokine release, and histopathological damage to the organs as well as decreasing the expression of the KIM-1 gene, which is an indicator of kidney damage.

Hyperbaric oxygen administration mitigates the expression of kidney injury molecule-1, inflammatory cytokines, and apoptotic markers during cold ischemic time in kidney transplantation

The aim of this study was to investigate the effect of hyperbaric oxygen (HBO2) administration applied during cold ischemic time to organs removed from donors before kidney transplantation. A total of 24 rats were divided into three groups: Group 1 was the control group, Group 2 received 60 minutes of HBO2 at 2.5 atmospheres absolute, and Group 3 received 120 minutes of 2.5 ATA HBO2. The renal artery was entered with a polyethylene catheter and perfused with a standard organ preservation solution. Falcon tubes containing organs obtained from rats in Groups 2 and 3 were placed in a box supported by ice blocks. The temperature was kept constant at 4 °C and the box was placed in a pressure tank with 2.5 ATA HBO2. HBO2 was applied for 60 and 120 minutes, respectively. Organ samples were harvested at the end of 24 hours for histopathological evaluation, immunohistochemical analysis of TNF-α and IL-18, TUNEL analysis for apoptosis, and gene expression levels of kidney injury molecule-1 (KIM-1) and caspase-3. In histopathological examinations, hematoxylin and eosin staining was performed and samples were evaluated for tubular necrosis and vacuolization criteria. Group 2 and Group 3 had significant decreases compared to Group 1 in this regard. Immunohistochemical staining was performed for TNF-α, IL-18, and apoptosis levels; significant decreases were found in Groups 2 and 3. There were significant decreases in Groups 2 and 3 for KIM-1 and caspase-3 gene expression levels compared to Group 1, as well. Thus, it was demonstrated that during the cold ischemic time before kidney transplantation, HBO2 administration to organs removed from donors can reduce apoptotic cell numbers, inflammatory cytokine release, and histopathological damage to the organs as well as decreasing the expression of the KIM-1 gene, which is an indicator of kidney damage.

Study on the impact of the drug (doxorubicin)-loaded platelet microparticles on cancer cells; the Daudi and Vero cell lines

Platelet-derived microparticles (PMPs) may be used as drug carriers for cancer treatments. Doxorubicin with fluorescent properties as a chemotherapeutic drug is available to treat many cancers. This study aims to compare different drug loading methods into PMPs and evaluate their effects on the viability and apoptosis of cancer cells. The platelet concentrates were taken from the Iranian Blood Transfusion Organization. PMPs were isolated on day 5 of platelet storage, and doxorubicin was loaded into PMPs through incubation, cell-penetrating peptides (CPP), and sonication methods. The rate of drug loading into PMPs was measured by the flow cytometry method. Finally, the effect of the drug-loaded microparticles on Daudi and Vero cells was investigated by determining the survival percentage (MTT) and caspase-3 gene expression levels (Real-time PCR). The average fluorescence light calculated in each of the incubation, sonication, and CPP methods was determined to be 79.09% ± 11.37%, 47.48% ± 25.12%, and 56.69% ± 23.24%, respectively. After 48 and 72 h of treating the Vero cells with drug-loaded microparticles, the survival rate decreased, and the expression of the caspase-3 gene was increased. In Daudi cells, similar results were obtained after 72 h of treatments. Also, the incubation method provided a higher rate of drug loading into PMPs. Therefore, this method can be used for drug loading in PMPs. As a result, as drug carriers, PMPs can lower cell viability in cancer cells.

Investigation of CYP2E1 and Caspase-3 Gene Expressions in COVID-19 patients

BackgroundCOVID-19 pandemic spread around the world like an infectious disease that presents waved effects on patients. Some patients needed ICU and respiratory support. Some patients only had flu-like symptoms. Cytokine storm and elevated ROS were serious problems for treatment. Apoptotic genes and CYP Family are part of these mechanisms. AimIn this study, our aim was to examine the gene expression CYP2E1 and Caspase-3 in patients with COVID-19 infection. Method60 COVID-19(+) patients (ICU and non-ICU patients) and 30 healthy volunteers were enrolled to study. To measure the level of gene expression qPCR was used. The 2-ΔΔCt method was utilized to analyze gene expression. ResultsThe expression of CYP2E1 and Caspase-3 genes showed a significant discrepancy between patients and healthy individuals. Caspase-3 expression increased (p=0,0041) but CYP2E1 expression decreased (p=0,0214) in COVID-19 patients compared to healthy individuals. Both levels of gene expression were lower in patients with affected lungs than patients with unaffected lungs (p<0,05). Laboratory findings including d-Dimer, LDH, platelet count, lymphocyte count were related to both gene expressions (p<0,05). We found no correlation between CYP2E1 and Caspase-3 expressions. ConclusionThe expression of Caspase-3 demonstrated apoptotic situations of patients but was not related to the CYP2E1 expression level. CYP2E1 gene expression is an important actor to metabolize endogens and xenobiotics however, COVID-19 patients demonstrated decreased CYP2E1 expression. CYP2E1 and Caspase-3 gene expression levels may be used as a diagnostic tool for COVID-19 patients.

The evaluation of both the expression and serum protein levels of Caspase-3 gene in patients with different degrees of SARS-CoV2 infection.

To evaluate the effects of Caspase‐3 (CASP3) gene expression and serum levels on preventing severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) infection. A total of 41 individuals (male: 21; female: 20) with SARS‐CoV‐2 infection were included in the current study. Hemograms were examined from patient blood samples, and CASP3 gene expression levels were detected. Also, human CASP3 levels were determined from the serum samples of patients. The mean age of patients was 56.220 ± 18.937 years. Significant differences were detected among all groups for CASP3 2−ΔΔCt (p = 0.014) and CASP3 concentration (p = 0.024). The relationship between CASP3 2−ΔΔCt levels and hemoglobin (p = 0.023), between CASP3 2−ΔΔCt levels and C‐reactive protein (CRP) (p = 0.001), between CASP3 2−ΔΔCt levels and ferritin (p = 0.003), between CASP3 2−ΔΔCtlevels and lactate dehydrogenase (p = 0.001), and between CASP3 2−ΔΔCt levels and SpO2 (p = 0.006) were statistically significant. Also, the relationship between CASP3 concentration levels and SpO2 was statistically significant (p < 0.046). The CASP3 gene and/or its products have an important function to prevent injury caused by SARS‐CoV‐2 infection. They play crucial roles in maintaining cellular homeostasis and viability. Perhaps CASP3 levels may provide information about the severity of the disease.

Fucoidan Modulated Oxidative Stress and Caspase-3 mRNA Expression Induced by Sulfoxaflor in the Brain of Mice.

The current study aimed to investigate the role of fucoidan in the oxidative and apoptotic effects of sulfoxaflor, a neonicotinoid sulfoximine insecticide, in the brain of Swiss albino mice (Mus musculus). Sulfoxaflor and fucoidan were administered to mice at doses of 15mg/kg/day (1/50 oral LD50) and 50mg/kg/day, respectively, by oral gavage for 24h or 7days. The tGSH, TBARSand protein levels, and GPx, GR, and GST enzyme activities were determined by spectrophotometric methods. Caspase-3 gene expression level was determined by RT-PCR. Data analysis showed that brains of sulfoxaflor-treated mice exhibited higher TBARS levels; GPx, GR, and GST enzyme activities; and caspase-3 expression levels, as well as lower levels of tGSH. Co-administration of fucoidan and sulfoxaflor reduced the TBARS levels, increased tGSH levels, and increased GPx, GR, and GST enzyme activities. Fucoidan also decreased the sulfoxaflor-induced up-regulation of caspase-3 mRNA expression. Results of the present study showed that sulfoxaflor caused oxidative stress by inducing lipid peroxidation and altering GSH-dependent antioxidants in the brain of mice. In addition, sulfoxaflor may trigger apoptotic cell death shown by the up-regulation of caspase-3. Fucoidan treatment modulated all the aforementioned alterations in the brain of mice. It was concluded that fucoidan might have antioxidant effects that support the GSH-dependent antioxidant system and can play a modulator role in oxidative stress and caspase-3 expression in the brain of sulfoxaflor treated-mice.

Effect of Hydroalcoholic Extract of Ephedra major, Momordica charantia, and Resveratrol on Cytotoxicity and Caspase-3 Genes Expression Level in MCF-7 Breast Cancer Cell Line

Background: To increase the therapeutic effect of drugs to combat diseases, combination therapy with current chemical drugs and new medicines derived from medicinal plants is necessary. Objectives: The present work aimed to investigate the effect of hydroalcoholic extract of two medicinal plants, Ephedra major and Momordica cacharantia (Carla), and resveratrol drug on cell viability and expression levels of caspase-3 gene in MCF-7 cell line. Methods: In this experimental study, the hydroalcoholic extraction of tested plants was done with a Soxhlet extractor. The MTT assay and real-time PCR were used to determine cell toxicity and caspase-3 gene expression levels, respectively. Results: The highest and lowest cytotoxic effects of plant extracts and resveratrol were observed at concentrations of 500 and 150 µg/mL, respectively. The highest level of the caspase-3 gene expression was observed after 72 h of incubation by different concentrations of plant extracts and resveratrol. Conclusions: It can be concluded that both plant extracts could influence cell viability in MCF-7 cells via the increase of cell toxicity and expression of caspase-3 gene. Thus, these species could be used in the pharmaceutical industry.

JNK Signaling Pathway Suppresses LPS-Mediated Apoptosis of HK-2 Cells by Upregulating NGAL.

Objective To explore the role of the c-Jun N-terminal kinase (JNK) signaling pathway in upregulated NGAL expression and its antiapoptotic mechanism in lipopolysaccharide (LPS)-mediated renal tubular epithelial cell injury. Methods In vitro, HK-2 cells were divided into five groups (Con, LPS 1 h, LPS 3 h, LPS 6 h, and LPS 12 h groups) based on the time of LPS (10 μM) treatment. NGAL and caspase-3 gene expression levels were detected by RT-PCR to assess dynamic changes. HK-2 cells were pretreated with SP600125 (20 μM) for 2 hours, followed by LPS (10 μM) stimulation for 3 hours. NGAL and caspase-3 gene expression levels were then determined. Results NGAL mRNA was increased significantly within 6 hours, and caspase-3 mRNA was increased within 3 hours after treatment (P < 0.05). Correlation analysis showed a high correlation between their expression (r = 0.448, P < 0.05). After pretreatment with SP600125, mRNA expression of NGAL in the LPS group was inhibited, while that of caspase-3 was increased significantly. The NGAL mRNA expression level in the SB + LPS group was decreased significantly compared with that in the LPS group, but it was slightly higher than that in the SP group (∼1.5 times of that in the Con group). However, caspase-3 mRNA expression was increased significantly in the SB + LPS group (P < 0.001) (3.5 times of that in the Con group). It also showed a significant increase compared with SP and LPS groups (P < 0.001 vs. SB group; P < 0.05 vs. LPS group). We also found that NGAL and caspase 3 proteins were increased significantly in LPS and SP + LPS groups, but SP600125 decreased the NGAL level by almost 35% and increased the caspase 3 level by 50% in the SP + LPS group compared with the LPS group (P < 0.05). Conclusions The JNK signaling pathway inhibits LPS-mediated apoptosis of renal tubular epithelial cells by upregulating NGAL.

Zinc Protects against MDMA-Induced Apoptosis of Sertoli Cells in Mouse via Attenuation of Caspase-3.

Background3,4-Methylenedioxymethamphetamine (MDMA) disrupts function of the endocrine system and dif- ferent organs such as heart, blood vessels, kidney, liver and nervous systems. This study was conducted to evaluate impact of MDMA on apoptosis and Zinc in the MDMA-induced apoptosis of cultured Sertoli cells by measuring Caspase-3 gene expression.Materials and MethodsIn this experimental study, Sertoli cells were incubated with MDMA (0, 0.5, 1, 3, 5 mM), Zinc (0, 8, 16, 32, 64 μM) and Zinc (8 μM) prior to adding MDMA (5 mM) for 24 and 48 hours. MTT assay was used for evaluating impacts of these conditions on the viability of Sertoli cells. Caspase-3 gene expression level was detected using quantitative reverse transcription PCR (qRT-PCR) in all of the tested groups.ResultsFinding showed that cellular viability was decreased and level of Caspase-3 mRNA was increased in MDMA treated cells. Additionally, pre-treatment with Zinc (8 μM) attenuated MDMA-induced apoptosis and down-regulated caspase-3. The mean of caspase-3 mRNA level (fold change ± SE) was 3.98 ± 1.18, 0.31 ± 0.28, and 1.72 ± 0.28 in re- spectively MDMA (5 mM), Zinc (8 μM), and Zinc+MDMA groups vs. control group. The mean of Caspase-3 mRNA (fold change) was not statistically different in the tested groups (P>0.05), unless MDMA (5 mM) group (P = 0.008).ConclusionWe suggest that MDMA toxicity could be involved in apoptosis of Sertoli cells. In addition, Zinc could reduce MDMA-induced apoptosis by down-regulation of Caspase-3 mRNA levels.

In vitro evaluation of the effects of capsaicin on normal and cancerous cells of human cartilage.

Chondrosarcoma is a common form of bone cancer which effects the fibrous connective tissue around a joint. It most commonly develops in legs, arms, shoulder blades, rib cage, and pelvis. Capsaicin is an active bitter compound found in red pepper, the fruit of the species Capsicum annuum, and it has been shown to have a lethal effect on different types of cancer. However, to date, investigation of its effect on human chondrosarcoma cells has remained limited. In the study presented here, we determined IC50 values of capsaicin for chondrosarcoma and chondrocyte cells in both fetal bovine serum (FBS)-containing and FBS-deprived media, and no statistically significant difference was found between the cell types. Besides, when the cells were cultured with capsaicin at their determined IC50 value for 24 h and their caspase-3 gene expression levels were detected by real-time polymerase chain reaction (RTPCR) and western blotting, it was demonstrated that the caspase-3 protein and mRNA levels were not altered in any cells upon capsaicin exposure, suggesting a caspase-independent pathway for cell death. Migration and invasion abilities of the cancerous cells, on the other hand, were observed to decrease dramatically when the cells were exposed to capsaicin (P < 0.05).

Combined effects of Lenvatinib and iodine-131 on cell apoptosis in nasopharyngeal carcinoma through inducing endoplasmic reticulum stress.

Nasopharyngeal carcinoma (NPC) is a type of malignant tumor characterized by high invasiveness, metastatic potential and worldwide incidence among patients with head and neck cancer. It has previously been demonstrated that Lenvatinib (LEB) is an efficient anti-cancer agent by multi-targeting of tyrosine kinase inhibitors. Iodine-131 (I-131) therapy has been accepted for the treatment of thyroid cancer and other carcinomas. In the present study, the combined effects of LEB and I-131 were investigated on NPC and the potential signal pathway mediated by LEB and I-131 on NPC cells was explored. Inhibitory effects of LEB and I-131 for NPC cells growth were investigated via MTT assay. Migration and invasion of NPC cells was studied by aggression assays following incubation of LEB and I-131. Apoptosis of NPC cells and tissues were analyzed via flow cytometry and TUNEL assay, respectively. Apoptosis-related gene expression levels in NPC cells following treatment with LEB and I-131 were determined by western blotting. Endoplasmic reticulum (ER) stress in NPC cells were analyzed in NPC cells and tumor tissues. Immunohistochemistry was used to analyze the efficacy of LEB and I-131 in NPC-tumor bearing mice. The results demonstrated that combined treatment of LEB and I-131 significantly inhibited growth, apoptosis, migration and invasion of NPC cells compared with single agent therapy. Apoptosis-related gene expression levels of caspase-3 and caspase-9 were upregulated by LEB and I-131, whereas B call lymphoma-2, and P53 were downregulated in NPC cells and tumor tissues. In addition, signal mechanism analysis demonstrated that combined treatment of LEB and I-131 promoted expression levels of activating transcription factor 6, inositol-requiring protein 1 (IER1), protein kinase RNA-like endoplasmic reticulum kinase (RERK), and C/EBP homologous protein in NPC cells. Furthermore, combined treatment of LEB and I-131 markedly inhibited in vivo growth of NPC and further prolonged survival of experimental mice compared with single agent and control groups. Immunohistochemistry indicated that c-jun N-terminal kinase and Caspase-3 were increased in NPS tumor tissues in xenograft models treated with LEB and I-131. Apoptotic bodies were also increased in tumors treated by LEB and I-131. In conclusion, these findings indicate that combined treatment of LEB and I-131 may inhibit NPC growth and aggression through upregulation of ER stress, suggesting combined treatment of LEB and I-131 may be a potential therapeutic schedule for the treatment of NPC.

Design and synthesis of novel pyrrolo[2,3-a]carbazoles: 7-Chloro-2-oxo-3a-(2′-oxo-2′,3′-dihydro-1′H-indol-3′-yl)-2,3,3a,4,5,10-hexahydro-pyrrolo[3,2-a]carbazole-1-carbonitrile as an efficient anticancer agent

Highly efficient poly functionalized pyrrolo[3,2-a]carbazoles via ring contraction through rearrangement and intramolecular Michael addition reaction using one pot multicomponent reaction (MCR) is reported for the first time. Free radical scavenging and anticancer activities were determined by DPPH and MTT assays respectively. Of these, compound 8d exhibited most potent activity against HCT-15 human colon cancer cell lines with an IC50 value of 9.9 μM and low toxicity toward normal human red blood cells. The morphological changes were visualized using scanning electron microscopy (SEM) technique, intracellular ROS generation measured by spectrofluorometer and gene expression levels of caspase-3, caspase-9 and Bcl-2 were determined using Semi quantitates PCR analysis for the target compound. Further, the structure activity relationships were also carried out. The results of the present study revealed that among pyrrolo[3,2-a]carbazole compounds, 7-chloro-2-oxo-3a-(2′-oxo-2′,3′-dihydro-1′H-indol-3′-yl)-2,3,3a,4,5,10-hexahydro-pyrrolo[3,2-a]carbazole-1-carbonitrile could be exploited as an excellent anticancer agent against colon cancer cells.

The effect of exposure to 1800 MHz radiofrequency radiation on epidermal growth factor, caspase-3, Hsp27 and p38MAPK gene expressions in the rat eye.

Radiofrequency electromagnetic fields (RF-EMF) may induce DNA damage and oxidative stress in human lens epithelial cells (LECs). We aimed to investigate the expression levels of heat shock protein 27 (Hsp27), p38 mitogen-activated protein kinase (p38MAPK), epidermal growth factor receptor (EGFR) and caspase-3 gene expression levels in rat eye that was exposed to 1800 MHz RF-EMF. Thirty-seven female Wistar albino rats were divided into three groups. The rats in the study group (n = 9) were exposed to 1800 MHz RF-EMF at an electric field 6.8 ± 0.1 V/m and 0.06 W/kg specific absorption rate (SAR) for 2 hours per day for eight weeks. Sham group (n = 9) was kept under similar conditions as the exposed group without exposure to RF-EMF. The rats in all three groups were sacrificed and their eyes were removed. Hsp27, p38MAPK, EGFR, caspase-3 gene expression levels were investigated in detail with real-time polymerase chain reactions (Real-Time PCR). caspase-3 and p38MAPK gene expression were significantly upregulated in the ocular tissues following exposure to RF-EMF (p < 0.05). According to our findings, eye cells recognize EMF as a stress factor, and in response, activate caspase-3 and p38MAPK gene expressions. These results confirm that RF-EMF can cause cellular damage in rat ocular cells (Tab. 2, Fig. 3, Ref. 37).

Dietary PUFA Increase Apoptosis in Stomach of Patients with Dyspeptic Symptoms and Infected with H. pylori

Drug-resistant strains of Helicobacter pylori and poor treatment response are the main reasons for the failure in eradicating it in patients. Polyunsaturated fatty acids (PUFA) have an inhibitory effect on bacterial growth. The aim of this study was to investigate the effect of PUFA in combination with standard triple therapy on apoptosis in H. pylori infected subjects with dyspeptic symptoms. This study was a double-blind clinical trial in which 34 H. pylori infected subjects with dyspeptic symptoms were randomly divided into two groups of 17 patients. The control group received standard triple therapy (amoxicillin, clarithromycin and omeprazole) and the experimental group received the standard therapy and PUFA for two weeks. Gene expression levels of caspase-3, BCL-2 and Bad proteins were studied with real-time PCR, while protein levels were quantified in frozen sections and using immunohistochemistry. Compared with the control group, a significant increase (p<0.01) was observed in the expression of caspase-3 and Bad genes and a significant reduction (p<0.05) in the expression of Bcl-2 gene. The protein level of active caspase-3 and Bad protein was significantly increased and the level of Bcl-2 protein was significantly decreased (p<0.05). The results of this study show that oral administration of PUFA in combination with the standard triple therapy increased apoptosis in H. pylori-infected patients with dyspeptic symptoms. This increase in apoptosis may partly reduce drug resistance in these patients. Our results suggest inclusion of a dietary PUFA containing fatty acid supplement may improve treatment of patients that are refractory to the standard triple therapy.

Effect Of Sprint Interval &amp; Eccentric Training On Gene Expression Levels Of Caspase3 &amp; Bax/bcl-2 In Rat Skeletal Muscle

PURPOSE: Apoptosis is a process of programmed cell death that occurs in different stress factors in the body. One of these stresses is exercise, exercise is a strong physiological stimulus which can influence a number of intracellular and extracellular signaling pathways. Till now, few researches shows the effect of sprint interval training and eccentric training on caspase3, bcl2 and bax gene expression changes. This study is exploring the effect of nine weeks of sprint interval training and eccentric training on caspase3, bcl2 and bax expression in soleus and SVL muscles METHODS: twenty four male Sprague Dawley rats purchased from Razi Institute were divided into three groups: control (n=8), sprint interval training (n=8) and eccentric training (n=8). Sprint interval training was included of one minute sprinting on animal treadmill with 2-4 minutes rest, 6-10 sets per session on 5-6 days a week, and eccentric downhill running was included running on -16° slope with 16 m/min for 90 minutes which in both the intensity of the trainings during the nine weeks gradually increased. The evaluation of gene expression was done by Real time PCR method RESULTS: Considering the low number of samples (n=8) and the normality of data rejected by Shapiro-Wilk test, Mann Whitney teat for differential between training and control groups were used. Also, caspase3 expression increased in all groups. All of these differences were not significant except for soleus muscle of eccentric training group. Also Bax/bcl-2 ratio increased in all groups and All of these differences were not significant. CONCLUSIONS: we conclude that nine weeks of Sprint interval training could lead to a small increase in caspase3 expression, Bax/bcl-2 ratio as the influential factor of apoptosis in all groups, except a large increase in caspase3 expression and ultimately strong apoptosis for soleus muscle in eccentric training group. Furthermore, it should be mentioned that the type of training and muscle could influenced the amount of gene expression levels of caspase3, bax/bcl2.

Involvement of regucalcin gene promoter region-related protein-p117, a transcription factor, in human obesity.

Regucalcin gene promoter region-related protein-p117 (RGPR-p117; gene symbol, rgpr-117) was identified in 2001 as a novel transcription factor that specifically binds to a nuclear factor I consensus motif, TTGGC(N)6CC in the promoter region of the regucalcin (rgn) gene. The human RGPR-p117 gene consists of 26 exons spanning ~4.1 kbp and is localized on chromosome 1q25.2. The nuclear translocation of cytoplasm RGPR-p117 is mediated via the protein kinase C-dependent signaling pathway. Overexpression of RGPR-p117 enhances the transcription activity of rgn, and a protective effect on cell death by inhibition of gene expression levels of caspase-3, caspase-8 and FADD proteins that possess the TTGGC motif in the promoter region of those genes was revealed. RGPR-p117 has a crucial role as a transcription factor. Notably, RGPR-p117 was shown to localize in the plasma membranes, mitochondria and microsomes (endoplasmic reticulum; ER). RGPR-p117, which is located in the ER, was also shown to have a role as an ER export factor implicated in the transports of proteins and lipids. As a result of this finding, it was proposed in 2007 that RGPR-p117 is renamed SEC 16 homolog B, endoplasmic reticulum export factor (SEC16B). Recently, there is increasing evidence that RGPR-p117/SEC16B may be involved in human obesity. Thus, the current review presents data regarding the involvement of RGPR-p117 in human obesity.

Scabraside D Extracted from Holothuria scabra Induces Apoptosis and Inhibits Growth of Human Cholangiocarcinoma Xenografts in Mice.

Scabraside D, a sulfated triterpene glycoside extract from sea cucumber Holothulia scabra, shows various biological activities, but effects on human cholangiocarcinoma cells have not previously been reported. In the present study, we investigated the activity of scabraside D against human cholangiocarcinoma (HuCCA) both in vitro and for tumor growth inhibition in vivo using a xenograft model in nude mice. Scabraside D (12.5-100 μg/mL) significantly decreased the viability and the migration of the HuCCA cells in a dose-dependent manner, with 50% inhibitory concentration (IC50) of 12.8 ± 0.05 μg/mL at 24 h. It induced signs of apoptotic cells, including shrinkage, pyknosis and karyorrhetic nuclei and DNA fragmentation on agarose gel electrophoresis. Moreover, by quantitative real-time PCR, scabraside D effectively decreased Bcl-2 while increasing Bax and Caspase-3 gene expression levels suggesting that the scabraside D could induce apoptosis in HuCCA cells. In vivo study demonstrated that scabraside D (1 mg/kg/day, i.p. for 21 days) significantly reduced growth of the HuCCA xenografts without adverse effects on the nude mice. Conclusively, scabraside D induced apoptosis in HuCCA cells and reduced the growth of HuCCA xenographs model. Therefore, scabraside D may have potential as a new therapeutic agent for cholangiocarcinoma.

Reversal effect of monoisoamyl dimercaptosuccinic acid (MiADMSA) for arsenic and lead induced perturbations in apoptosis and antioxidant enzymes in developing rat brain

Oxidative stress (OS) has been implicated in the pathophysiology of many neurodegenerative disorders. Several studies have shown that exposure to arsenic (As) and lead (Pb) produces oxidative stress, one of the most noted molecular mechanisms for the neurotoxicity of these metals. In the present study, we examined the effect of combined exposure to these metals (As and Pb) on the activity levels of antioxidant enzymes and apoptotic marker enzymes in brain regions (cerebral cortex, hippocampus and cerebellum) of rats at postnatal day (PND) 21, 28 and 3 months age and compared the toxicity levels with individual metals (As or Pb). Further, we also evaluated the therapeutic efficacy of a chelating agent, monoisoamyl dimercaptosuccinic acid (MiADMSA) against arsenic and lead induced developmental neurotoxicity. Pregnant rats were exposed to sodium meta-arsenite (50ppm) and lead acetate (0.2%) individually, and in combination (As=25ppm+Pb=0.1%) via drinking water throughout perinatal period (GD 6 to PND 21). MiADMSA (50mg/kg, orally through gavage) was given for three consecutive days to the PND 18 pups (i.e., PND 18 to PND 20). Exposure to metal mixture resulted in a significant decrease in the activity levels of antioxidant enzymes such as manganese-superoxide dismutase (Mn-SOD), Cu/Zn superoxide dismutase (Cu/Zn-SOD), catalase (CAT) and glutathione peroxidase (GPx) while the malondialdehyde (MDA) levels and mRNA expression levels of caspase-3 and caspase-9 were significantly increased in all the three brain regions. The observed alterations were greater with exposure to metal mixture than individual metals (As or Pb) and the changes were more prominent at PND 28 and greater in cerebral cortex than hippocampus and cerebellum. Interestingly, chelation therapy with MiADMSA showed significant recovery in antioxidant enzymes, lipid peroxidation and gene expression levels of caspase-3 and caspase-9. From these findings, it can be concluded that combined exposure to As and Pb showed an additive effect on antioxidant enzymes than individual metal exposure and chelation therapy with MiADMSA significantly reversed the As and Pb induced apoptosis and oxidative stress, a major contributing factor to neurotoxicity.

Antitumor effects of ginsenoside Rg3 on human hepatocellular carcinoma cells

The antitumor effects of ginsenoside Rg3 have been reported in several kinds of human malignant tumors. The purpose of this study was to investigate whether ginsenoside Rg3 can inhibit the growth of human hepatocellular carcinoma cell lines and to discuss the possible molecular mechanism(s). We cultured the human hepatocellular carcinoma cell lines, SMMC-7721 and HepG2. The cells were treated with different concentrations of ginsenoside Rg3 (0, 25, 50, 75 and 100 µg/ml), and the cell proliferation was detected by MTT assay at the 12, 24, 36 and 48 h time-points. Flow cytometry experiments were carried out to investigate the effect of Rg3 on cell apoptosis after the cells had been treated with Rg3 (50 and 100 µg/ml) for 24 and 48 h. The expression levels of caspase-3, bax and bcl-2 in Rg3-treated cells (100 µg/ml, 48 h), as well as normal cells were detected through real-time PCR experiments. MTT assay showed that the inhibition rate of cell proliferation in the Rg3 groups was significantly higher compared to the control groups in both the SMMC-7721 and HepG2 cell lines, and the inhibition rate increased with increasing Rg3 concentrations and duration of treatment. Flow cytometry analysis demonstrated that the Rg3 groups had a significantly higher cell apoptotic rate compared to the control groups in both the SMMC-7721 and HepG2 cell lines, and that the effect of Rg3 on cell apoptosis occurred in a concentration- and time-dependent manner, as was also shown by the MTT assay. Real-time PCR analysis showed that the gene expression levels of caspase-3 and bax were significantly enhanced in the Rg3 groups compared to the control groups in both the SMMC-7721 and HepG2 cell lines, but the gene expression level of bcl-2 was significantly inhibited. These results indicate that ginsenoside Rg3 can effectively inhibit the growth of human hepatocellular carcinoma cell lines by inhibiting cancer cell proliferation and promoting cancer cell apoptosis, and it may promote cancer cell apoptosis via the endogenous mitochondrial-mediated caspase-dependent apoptotic pathway.

The effects of caspase-3 siRNA on the neurobehavior of mice exposed to aluminum

To investigate the effects of caspase-3 siRNA on the neurobehavior of mice exposed to aluminum. Male KunMing mice (3 months old) were randomly divided into 4 groups by weight:blank control group (4 microl normal saline), Al group (4 microl 0.5% AlCl3), Al plus empty vector group(3 microl 0.5% AlCl3 plus control siRNA expression vector)and Al plus RNAi group (3 microl 0.5% AlCl3 plus targeted siRNA expression vector). All groups were treated by lateral cerebral ventricle micro-injection for 5 days. The neurobehavior was tested by the Morris water maze test, Open-field and Step-down tests for all treated mice. Pathological changes in hippocampus was observed by electron microscopy, the caspase-3 gene expression levels were detected using RT-PCR. The results of Step-down test indicated that as compared with control group, the latent time [LT, (44.67 +/- 10.60) s] in A1 group decreased significantly, the error number (3.63 +/- 0.52) in Al group increased significantly and the LT [(68.00 +/- 14.70) s] in Al plus empty vector group decreased significantly (P<0.05). the LT [(239.50 +/- 19.36) s] in Al plus RNAi group increased significantly and the error number in Al plus RNAi group decreased significantly, as compared with Al group (P<0.05). The results of Morris water maze test showed that as compared with control group, the LT in Al group increased significantly, and residence time in the former platform quadrant decreased significantly and the LT in Al plus empty vector group increased significantly (P<0.05). The LT in Al plus RNAi group was significantly longer than that in Al group (P<0.05). The results of open-field test demonstrated that as compared with control group, the time in the central grid in Al group and Al plus empty vector group increased significantly, the rearing number and the modification number in Al group and Al plus empty vector group decreased significantly (P< 0.05). As compared with Al group, the time in the central grid in Al plus RNAi group decreased, the inter-cell number, the rearing number and the modification number increased significantly (P<0.05). The results of electron microscopic examination exhibited that a slight change of hippocampal cells appeared in control group, the obvious pathological changes of hippocampal cells appeared in Al group and Al plus empty vector group, but the pathological changes of hippocampal cells in Al plus RNAi group significantly reduced as compared with Al group. The results of thionin staining indicated that the layers of neural cells of hippocampal CA3 were more clear and there was not obvious denatured injury of neural cells of hippocampal CA3 in control group. The number and Nissl body color of neural cells of hippocampal CA3 in Al group and Al plus empty vector group decreased significantly. After RNA interference, the number and Nissl body color of neural cells of hippocampal CA3 increased obviously. The expression levels of caspase-3 gene in Al group and Al plus empty vector group were 2.24 +/- 0.57 and 2.28 +/- 0.33, respectively, which were significantly higher than that (1.00 +/- 0.00) in control group (P<0.05). The expression level of caspase-3 gene in Al plus RNAi group was 0.44 +/- 0.08, which was significantly lower than those in Al group and control group (P<0.05). Aluminum can decrease the learning and memorizing ability, and inhibited the activity or exploration function of mice. It is suggested that Caspase-3 siRNA may reduce the neurotoxicity induced by aluminum to a certain extent.