Number Of Apoptotic Bodies Research Articles

Human endometrium-derived mesenchymal stem/stromal cells application in endometrial-factor induced infertility.

Endometrial-factor induced infertility remains one of the most significant pathology among all fertility disorders. Stem cell-based therapy is considered to be the next-generation approach. However, there are still issues about successfully retrieving human endometrium-derived mesenchymal stem/stromal cells (hEnMSCs). Moreover, we need to establish a better understanding of the effect of hEnMSCs on the endometrial recovery and the clinical outcome. According to these challenges we created a multi-step study. Endometrium samples were collected from females undergoing assisted reproductive technology (ART) procedure due to couple infertility. These samples were obtained using an endometrium scratching. The hEnMSCs were isolated from endometrium samples and characterized with flow cytometry analysis. Groups of endometrium injured female mice were established by the mechanical injury to uterine horns and the intraperitoneal chemotherapy. The hEnMSCs suspension was injected to some of the studied female mice at approved time intervals. Histological changes of mice uterine horns were evaluated after Masson's trichrome original staining, hematoxylin and eosin (H&E) staining. The fertility assessment of mice was performed by counting formed embryo implantation sites (ISs). The expression of fibrosis related genes (Col1a1, Col3a1, Acta2, and CD44) was evaluated by the reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Results showed that endometrium scratching is an effective procedure for mesenchymal stem/stromal cells (MSCs) collection from human endometrium. Isolated hEnMSCs met the criteria for defining MSCs. Moreover, hEnMSCs-based therapy had a demonstrably positive effect on the repair of damaged uterine horns, including a reduction of fibrosis, intensity of inflammatory cells such as lymphocytes and polymorphonuclear cells (PMNs) and the number of apoptotic bodies. The injured mice which recieved hEnMSCs had higher fertility in comparison to the untreated mice. Gene expression was reflected in histology changes and outcomes of conception. In conclusion, hEnMSCs demonstrated a positive impact on endometrium restoration and outcomes of endometrial-factor induced infertility. Further exploration is required in order to continue exploring the multifactorial associations between stem cell therapy, gene expression, endometrial changes and reproductive health, so we can identify individually effective and safe treatment strategies for endometrial-factor induced infertility, which is caused by mechanical effect or chemotherapy, in daily clinical practise.

Involvement of the peroxisome proliferator-activated receptor gamma (Pparγ) and matrix metalloproteinases-2 and -9 (Mmp-2 and -9) in the mechanism of action of di(2-ethylhexyl)phthalate (DEHP) in cultured mouse brain astrocytes and neurons

Di(2-ethylhexyl)phthalate (DEHP) is one of the most widely used phthalates in industry. It has been shown that, after entering the body, DEHP has the ability to cross the blood-placenta and blood-brain barriers. One of the proposed mechanisms of action of DEHP is the activation of peroxisome proliferator-activated receptors (PPARs). Many different functions of PPARγ in cells have been demonstrated, one of which is the modulation of the activation of matrix metalloproteinases (MMPs). The aim of this study was to investigate the role of Pparγ, Mmp-2, and Mmp-9 in the mechanism of action of DEHP. The experiments were performed on in vitro primary murine neurons and astrocytes. The results showed that DEHP has a pro-apototic effect on neurons, causing an increase in caspase-3 activity and in the number of apoptotic bodies. However, in astrocytes, the increase in caspase-3 activity was not related to the apoptosis process, as no increase in the formation of apoptotic bodies was observed. Moreover, DEHP increased the proliferation of astrocytes, which was confirmed by the increase in the amount and expression of the Ki-67 protein. In astrocytes, DEHP decreased the expression of the Pparγ and Mmp-9 proteins but increased the expression of the Mmp-2 protein. In DEHP neurons, it increased the expression of the Pparγ protein but decreased the expression of the Mmp-2 and Mmp-9 proteins.

The Effect of Chlordiazepoxide Consumption on the Hippocampus of Neonatal Rats During Pregnancy

Background: Chlordiazepoxide is an anti-anxiety drug commonly used by young people and pregnant women to reduce anxiety. The adverse effects of this drug on cholinergic nervous system function have been demonstrated. Therefore, in this study, the effect of chlordiazepoxide consumption during pregnancy was evaluated on the rats infant hippocampus. Materials and Methods: Nine pregnant Wistar rats were randomly divided (n=3 per group) into control, experimental (daily intraperitoneal injection of chlordiazepoxide at a dose of 10 mg/kg for 21 days), and vehicle (same amount of normal saline) groups. Two weeks after birth, the neonate brains were removed from the skull and prepared for Nissl and TUNEL stainings. The expressions of pro- and anti-apoptotic genes were evaluated. Results: The number of healthy neurons in different areas of the neonatal hippocampus in the experimental group was significantly reduced compared to control and vehicle groups, and the number of apoptotic bodies was increased (P<0.05). However, there was no significant difference between the numbers of these cells in the control and vehicle groups. The expression of pro-apoptotic genes in the experimental group increased significantly compared to the control group instead of the expression of anti-apoptotic genes decreased significantly. Conclusion: Chlordiazepoxide during pregnancy can cause neuronal damage in the hippocampus of neonatal Wistar rats.

CTPS cytoophidia formation affects cell cycle progression and promotes TSN‑induced apoptosis of MKN45 cells.

Cytidine triphosphate synthase (CTPS) forms filamentous structures termed cytoophidia in numerous types of cell. Toosendanin (TSN) is a tetracyclic triterpenoid and induces CTPS to form cytoophidia in MKN45 cells. However, the effects of CTPS cytoophidia on the proliferation and apoptosis of human gastric cancer cells remain poorly understood. In the present study, CTPS-overexpression and R294D-CTPS mutant vectors were generated to assess the effect of CTPS cytoophidia on the proliferation and apoptosis of gastric cancer MKN45 cells. Formation of CTPS cytoophidia significantly inhibited MKN45 cell proliferation (evaluated using EdU incorporation assay), significantly blocked the cell cycle in G1 phase (assessed using flow cytometry) and significantly decreased mRNA and protein expression levels of cyclin D1 (assessed by reverse transcription-quantitative PCR and western blotting, respectively). Furthermore, the number of apoptotic bodies and apoptosis rate were markedly elevated and mitochondrial membrane potential was markedly decreased. Moreover, mRNA and protein expression levels of Bax increased and Bcl-2 decreased markedly in MKN45 cells following transfection with the CTPS-overexpression vector. The proliferation rate increased, percentage of G1/G0-phase cells decreased and apoptosis was attenuated in cells transfected with the R294D-CTPS mutant vector and this mutation did not lead to formation of cytoophidia. The results of the present study suggested that formation of CTPS cytoophidia inhibited proliferation and promoted apoptosis in MKN45 cells. These results may provide insights into the role of CTPS cytoophidia in cancer cell proliferation and apoptosis.

Propofol and Sevoflurane Alleviate Malignant Biological Behavior and Cisplatin Resistance of Xuanwei Lung Adenocarcinoma by Modulating the Wnt/β-catenin Pathway and PI3K/AKT Pathway.

Recent studies have shown that propofol and sevoflurane, two common anesthetics, can prevent tumor development. The prevalence of lung adenocarcinoma in China is highest in Yunnan Xuanwei, and many patients with lung cancer in this area are often resistant to platinum-based treatments. The objective of the study was to investigate the effects of propofol and sevoflurane on malignant biological behavior and cisplatin resistance of Xuanwei lung adenocarcinoma. Herein, XWLC-05/R, a cisplatin-resistant cell line of XWLC-05 cells from Xuanwei lung adenocarcinoma, was constructed. The XWLC-05 cells and XWLC-05/R cells were treated with propofol and sevoflurane singly or as a combination and subjected to CCK-8 assay, clone formation tests, and flow cytometry analysis to assess the proliferation level of cells. The morphology and number of apoptotic bodies in XWLC-05 cells and XWLC-05/R were examined with a transmission electron microscope. The ANNEXIN V-FITC/PI and transwell assays were performed to evaluate apoptosis, invasion, and migration of the cells. Subsequently, we constructed a Xuanwei lung adenocarcinoma xenograft model to investigate the effects of propofol and sevoflurane on the tumorigenicity of XWLC-05 cells in vivo. Treatment with propofol and sevoflurane significantly inhibited proliferation, invasion, migration, and induced apoptosis of XWLC-05 and XWLC-05/R cells. These effects were more pronounced when propofol and sevoflurane were co-incubated with the cells. Moreover, both propofol and sevoflurane significantly inhibited Wnt/β- catenin and PI3K/AKT pathways. Moreover, the two drugs effects suppressed the malignant biological behavior of XWLC-05 cells and XWLC-05/R cells, and this effect was counteracted by 740Y-P (PI3K/AKT pathway activator) and Wnt pathway activator 1 (Wnt/β-catenin pathway activator). In vivo experiments confirmed that propofol and sevoflurane alleviated the tumorigenicity of XWLC-05 cells. In summary, this study shows, for the first time, that propofol and sevoflurane decrease the proliferation, invasion, migration and induce apoptosis of XWLC-05 cells and XWLC-05/R cells by impeding the Wnt/β-catenin and PI3K/AKT signaling pathways, thereby alleviating the development of Xuanwei lung adenocarcinoma in vivo. Moreover, these effects are more pronounced when the two drugs are combined.

Novel amphibian-derived antioxidant peptide protects skin against ultraviolet irradiation damage.

Given the adverse impact of ultraviolet irradiation on human skin, as well as currently limited interventions, the discovery of new molecules with anti-photodamage potency remains critical. In this research, we obtained a new bioactive peptide (named OS-LL11, amino acid sequence 'LLPPWLCPRNK') from Odorrana schmackeri. Results showed that OS-LL11 could directly scavenge free radicals and sustain the viability of mouse keratinocytes challenged by ultraviolet B (UVB) irradiation or hydrogen peroxide (H2O2) by decreasing the levels of lipid peroxidation, malondialdehyde, and reactive oxygen species while increasing the level of catalase, Keap-1, HO-1, GCLM, and NQO1. Interestingly, topical application of OS-LL11 protected mouse skin against UVB irradiation damage by up-regulating the levels of superoxide dismutase, glutathione, and nitric oxide, but down-regulating the levels of H2O2, IL-1α, IL-1β, IL-6, TNF-α, 8-OHdG, Bcl-2, and Bax, as well as the number of apoptotic bodies. Our research demonstrated the anti-photodamage activity of a novel amphibian-derived peptide and the potential underlying mechanisms related to its free radical scavenging ability and antioxidant, anti-inflammatory, and anti-apoptotic activities. This study provides a new molecule for the development of anti-skin photodamage drugs or cosmetics and highlights the prospects of amphibian-derived peptides in photodamaged skin intervention.

Structural and molecular markers of aging heart: Sechenov University classification

Abstract Background The process of aging at the level of the cells is connecting with arising of senescence cell, that are characterized by several possible signs, including damage to the mitochondria and contraction of telomeres, epigenetic changes, impaired regulation of protein metabolism, sensitivity to nutrients, mitochondrial dysfunction, stem cell collapse, impaired intercellular communication. Postmitotic cells may also develop senescence and are called amitosenescence cells (AMCs). Goal To investigate structural and molecular characteristic of aging heart with special identification of AMCs and creating classification of aging heart status. Methods Study was performed on 52 biopsies (endomyocardial and operative) of the heart and 48 autopsy cases (38 women and 62 men) with diagnosis of dilated cardiomyopathies, chronic ischemic heart disease, hypertensive cardiomyopathies, rheumatic heart disease and their combination. All patients were divided in two groups according to the age of patients: 1 group included 80 patients (average age 72.9±13.3 years) and 2 control group - 20 patients (average age 42.9±13.3 years). Serial paraffin sections were stained with hematoxylin and eosin, picrofuchsin; p16ink, Apocas, CD68 and CD 45 were detected by immunohistochemistry (DakoCytomation). We have calculated percent of AMC, Apo and RCB in 30 fields of vision on X400 magnification. Results AMCs were found in the myocardium of 77 aging patients and only in 2 young patients (premature aging?). AMCs were characterized by prominent degenerative changes, p16ink expression and accumulation of lipofuscin. Reduction of capillary bad (RCB), increased number of apoptotic bodies and presence of CD68+ macrophages and CD45+ lymphocytes were found in stroma. There were three aging heart status degree: 1 – presence less than 15% of AMCs, 0,1% of Apo and RCB –less than 15% (20 patients); 2 – presence 16–30% of AMCs, 0,2- 1% of Apo and RCB – 16–30% (35); 3 – presence more than 31% of AMCs, 1–2% of Apo and RCB – more than 31% (27). Conclusion Aging heart has increased number of amitosenecence cells that results severity and chronicity of heart diseases in aging patients. Funding Acknowledgement Type of funding sources: None.

Electroacupuncture improves cognitive function by inhibiting NF-κB activity in rats with chronic fatigue syndrome

To observe the effect of electroacupuncture (EA) on the expression of NF-κB p65 in hippocampus and the morphology of hippocampus in rats with chronic fatigue syndrome (CFS), so as to explore its mechanism in improving cognitive dysfunction of CFS. Forty-eight SD rats were randomly divided into control, model, EA and inhibitor groups (n=12 in each group). The CFS model was established by multi-factor compound stress stimulation method. Rats of the EA group received EA (50 Hz, 1 mA) at "Baihui" (GV20), Emotional Area I and bilateral Sensory Area for 30 min, once daily for 15 days. For rats in the inhibitor group, pyrrolidine dithiocarbamate (100 mg·kg-1·d-1) was injected intraperitoneally, once a day for 15 days. Learning and memory ability was evaluated by Morris water maze test. HE staining was used to observe the morphology of hippocampus. Western blot was used to determine the expression level of NF-κB p65 in hippocampus. After mode-ling, the general status score was increased （P<0.01）, the escape latency was prolonged(P<0.01), the times of crossing the platform was decreased(P<0.01), and the expression level of NF-κB p65 in hippocampus tissue was significantly increased (P<0.05) in the model group compared with the control group. Compared with the model group, the general status score was decreased （P<0.01）, the escape latency was shortened(P<0.01), the times of crossing the platform was increased(P<0.01), and the expression level of NF-κB p65 in hippocampus tissue was significantly decreased (P<0.05) in the EA and inhibitor groups. HE staining showed that in the model group, the hippocampal nerve cells were arranged disorderly, the structure was loose, and the number of apoptotic bodies and inflammatory cells was significantly increased. The degree of tissue damage of the EA and inhibitor groups was milder than that of the model group. EA can improve the cognitive function in CFS rats, which may be associated with its effect in inhibiting the expression of NF-κB and reducing the inflammation response in hippocampus.

Silymarin inhibits proliferation of human breast cancer cells via regulation of the MAPK signaling pathway and induction of apoptosis.

Silymarin is a purified mixture of four isomeric flavonoids extracted from the seeds and fruit of the milk thistle plant, Silybum marianus (L.). Silymarin exhibits a wide variety of biological effects and is commonly used in traditional medicine. Therefore, the anticancer effects of silymarin on human breast cancer cells were investigated to determine its pharmacological mechanisms in vitro and in vivo. The viability and proliferation of MDA-MB- 231 and MCF-7 breast cancer cells were investigated using MTT and wound healing assays. Silymarin decreased the viability and proliferation of MDA-MB-231 and MCF-7 cells in a concentration-dependent manner. The number of apoptotic bodies, as shown by DAPI staining, was increased in a concentration-dependent manner, indicating that silymarin induces apoptosis. Additionally, changes in the expression levels of apoptosis-related proteins were demonstrated in human breast cancer cells using western blotting. Silymarin increased the levels of Bax, cleaved poly-ADP ribose polymerase, cleaved caspase-9 and phosphorylated (p-)JNK, and decreased the levels of Bcl-2, p-P38 and p-ERK1/2. Furthermore, the inhibitory effects of silymarin on MCF-7 tumor growth were investigated. In mice treated with silymarin for 3 weeks (25 and 50 mg/kg), MCF-7 tumor growth was inhibited without organ toxicity. In MCF-7 tumors, silymarin induced apoptosis and decreased p-ERK1/2 levels, as assessed using a TUNEL assay and immunohistochemistry. These results indicated that silymarin inhibited breast cancer cell proliferation both in vitro and in vivo by modulating the MAPK signaling pathway. Therefore, silymarin may potentially be used as a chemo-preventive or therapeutic agent.

Goji berry (Lycium barbarum L.) juice reduces lifespan and premature aging of Caenorhabditis elegans: Is it safe to consume it?

Goji berry fruit is considered a healthy food. However, studies on its effects on aging and safety are rare. This study is the first to evaluate the effects of goji berry juice (GBJ) on oxidative stress, metabolic markers, and lifespan of Caenorhabditis elegans. GBJ caused toxicity, reduced the lifespan of C. elegans by 50%, and increased the reactive oxygen species (ROS) production by 45–50% at all tested concentrations (1–20 mg/µL) of GBJ. Moreover, the highest concentration of GBJ increased lipid peroxidation by 80% and altered the antioxidant enzymes. These effects could be attributed to a pro-oxidant effect induced by GBJ polyphenols and carotenoids. Moreover, GBJ increased lipofuscin, glucose levels, number of apoptotic bodies, and lipase activity. The use of mutant strains demonstrated that these effects observed in the worms treated with GBJ were not associated with the Daf-16/FOXO or SKN-1 pathways. Our findings revealed that GBJ (mainly the highest concentration) exerted toxic effects and promoted premature aging in C. elegans. Therefore, its consumption should be carefully considered until further studies in mammals are conducted.

Characterization and function study of a glutamyl endopeptidase homolog from Nocardia seriolae.

Glutamic endopeptidases (Glu), belonging to the class of serine proteases, are a subfamily of chymotrypsin-like proteolytic enzymes, which are regarded as important virulence factors in bacteria. However, the roles of glutamic endopeptidases of Nocardia seriolae in pathogenic process still remain uncertain. Here, a glutamic endopeptidase homolog from N. seriolae (GluNS) was cloned and its function was elucidated. GluNS encoded a 414-aa protein which shared 93% identity to N. concava. In the phylogenetic tree, the glutamic endopeptidases of genus Nocardia clustered together firstly and then clustered with Streptomyces species. Moreover, GluNS was identified to be a secreted protein of N. seriolae and localized in the mitochondria of FHM cells. The transient overexpression of GluNS significantly induced increase in caspase-3 activity and decrease in ΔΨm values in FHM cells. The number of apoptotic bodies was remarkably higher than that in control group. Taken together, GluNS overexpression induced apoptotic characteristics in FHM cells. This study provided new insights into the function of glutamic endopeptidase from N. seriolae.

Protective Effects of Oenothera biennis against Hydrogen Peroxide-Induced Oxidative Stress and Cell Death in Skin Keratinocytes.

Background: Oenothera biennis (evening primrose) produces bioactive substances with a diverse range of pharmacological functions. However, it is currently unknown whether extract prepared from the aerial parts of O. biennis (APOB) can protect the skin against oxidative stress. Objective: The aim of this study is to investigate the protective effects of APOB against oxidative stress-induced damage in human skin keratinocytes (HaCaT) and elucidate the underlying mechanisms. Methods: We pretreated HaCaT cells with various concentrations of APOB or the antioxidant N-acetyl-L-cysteine before applying H2O2. We then compared the cell viability, intracellular reactive oxygen species (ROS) production, and DNA and mitochondrial damage between pretreated and untreated control cells using a range of assays, flow cytometry, and Western blot analysis and also examined the reducing power and DPPH free radical scavenging activity of APOB. Results: APOB pretreatment significantly increased cell viability, effectively attenuated H2O2-induced comet tail formation, and inhibited H2O2-induced phosphorylation of the histone γH2AX, as well as the number of apoptotic bodies and Annexin V-positive cells. APOB was found to have high reducing power and DPPH radical scavenging activity and also exhibited scavenging activity against intracellular ROS accumulation and restored the loss of mitochondrial membrane potential caused by H2O2. APOB pretreatment almost totally reversed the enhanced cleavage of caspase-3, the degradation of poly (ADP-ribose)-polymerase (PARP), DNA fragmentation that usually occurs in the presence of H2O2, and increased the levels of heme oxygenase-1 (HO-1), a potent antioxidant enzyme that is associated with the induction of nuclear factor-erythroid 2-related factor 2 (Nrf2). Conclusions: APOB can protect HaCaT cells from H2O2-induced DNA damage and cell death by blocking cellular damage related to oxidative stress via a mechanism that affects ROS elimination and by activating the Nrf2/HO-1 signaling pathway.

CNOT2 Is Critically Involved in Atorvastatin Induced Apoptotic and Autophagic Cell Death in Non-Small Cell Lung Cancers.

Though Atorvastatin has been used as a hypolipidemic agent, its anticancer mechanisms for repurposing are not fully understood so far. Thus, in the current study, its apoptotic and autophagic mechanisms were investigated in non-small cell lung cancers (NSCLCs). Atorvastatin increased cytotoxicity, sub G1 population, the number of apoptotic bodies, cleaved poly (ADP-ribose) polymerase (PARP) and caspase 3 and activated p53 in H1299, H596, and H460 cells. Notably, Atorvastatin inhibited the expression of c-Myc and induced ribosomal protein L5 and L11, but depletion of L5 reduced PARP cleavages induced by Atorvastatin rather than L11 in H1299 cells. Also, Atorvastatin increased autophagy microtubule-associated protein 1A/1B-light chain 3II (LC3 II) conversion, p62/sequestosome 1 (SQSTM1) accumulation with increased number of LC3II puncta in H1299 cells. However, late stage autophagy inhibitor chloroquine (CQ) increased cytotoxicity in Atorvastatin treated H1299 cells compared to early stage autophagy inhibitor 3-methyladenine (3-MA). Furthermore, autophagic flux assay using RFP-GFP-LC3 constructs and Lysotracker Red or acridine orange-staining demonstrated that autophagosome-lysosome fusion is blocked by Atorvastatin treatment in H1299 cells. Conversely, overexpression of CCR4-NOT transcription complex subunit 2(CNOT2) weakly reversed the ability of Atorvastatin to increase cytotoxicity, sub G1 population, cleavages of PARP and caspase 3, LC3II conversion and p62/SQSTM1 accumulation in H1299 cells. In contrast, CNOT2 depletion enhanced cleavages of PARP and caspase 3, LC3 conversion and p62/SQSTM1 accumulation in Atorvastatin treated H1299 cells. Overall, these findings suggest that CNOT2 signaling is critically involved in Atorvastatin induced apoptotic and autophagic cell death in NSCLCs.

Silymarin induces inhibition of growth and apoptosis through modulation of the MAPK signaling pathway in AGS human gastric cancer cells.

Apoptosis is regarded as a therapeutic target because it is typically disturbed in human cancer. Silymarin from milk thistle (Silybum marianum) has been reported to exhibit anticancer properties via regulation of apoptosis as well as anti-inflammatory, antioxidant and hepatoprotective effects. In the present study, the effects of silymarin on the inhibition of proliferation and apoptosis were examined in human gastric cancer cells. The viability of AGS human gastric cancer cells was assessed by MTT assay. The migration of AGS cells was investigated by wound healing assay. Silymarin was revealed to significantly decrease viability and migration of AGS cells in a concentration-dependent manner. In addition, the number of apoptotic bodies and the rate of apoptosis were increased in a dose-dependent manner as determined by DAPI staining and Annexin V/propidium iodide double staining. The changes in the expression of silymarin-induced apoptosis proteins were investigated in human gastric cancer cells by western blotting analysis. Silymarin increased the expression of Bax, phosphorylated (p)-JNK and p-p38, and cleaved poly-ADP ribose polymerase, and decreased the levels of Bcl-2 and p-ERK1/2 in a concentration-dependent manner. The in vivo tumor growth inhibitory effect of silymarin was investigated. Silymarin (100 mg/kg) significantly decreased the AGS tumor volume and increased apoptosis, as assessed by the TUNEL assay, confirming its tumor-inhibitory effect. Immunohistochemical staining revealed elevated expression of p-JNK and p-p38 as well as reduced expression of p-ERK1/2 associated with silymarin-treatment. Silymarin was revealed to reduce tumor growth through inhibition of p-ERK and activation of p-p38 and p-JNK in human gastric cancer cells. These results indicated that silymarin has potential for development as a cancer therapeutic due to its growth inhibitory effects and induction of apoptosis in human gastric cancer cells.

Neuroprotective Effect of Pentoxifylline on 3,4-Methylenedioxymethamphetamine-Induced Apoptosis in CA1 Cells of Wistar Rat Hippocampus.

Background: 3,4-Methylenedioxymethamphetamine is psychoactive and hallucinogenic and has been shown to produce neurotoxicity both in animals and in humans. Recently, vasodilator drugs such as pentoxifylline (PTX) have been introduced as an alternative with neuroprotective effects. There is no study about the protective effect of PTX on hippocampal apoptosis due to high-dose administration of 3,4-Methylenedioxymethamphetamine (MDMA), so in this study, the protective effect of PTX on the hippocampus of male Wistar rats following high-dose of the drug has been investigated. Materials and Methods: Twenty-four male Wistar rats weighing 250-300 g were randomly divided into four groups: control, sham (MDMA injection), experimental (MDMA+PTX injection), and vehicle (MDMA+saline) groups. Two weeks later, the brains were removed and prepared for TUNEL and western blot techniques. Concomitantly the hippocampus was removed to study the change in Bcl-2 and BAX mRNA expression with quantitative real-time polymerase chain reaction. Results: Data showed that the number of apoptotic bodies significantly decreased in the experimental group compared to the other groups, except for in control. Also, further investigation revealed that BAX reduced considerably, while Bcl-2 mRNA expression increased dramatically after PTX treatment. Conclusions: Our results suggest that PTX may be a neuroprotective agent, and its neuroprotective potential may contribute to reducing the severity of lesions in the hippocampus following a high dose administration of MDMA.

Abstract 4303: TanshinoneIIA, an isolated compound from Salvia miltiorrhiza, inhibits cell growth in colorectal cancer via activation of p38MAPK signaling pathway

Abstract Tanshinone IIA (TSIIA) is an active ingredient extracted from the root of Danshen (Salvia miltiorrhiza Bunge), a Chinese traditional herb. Recent studies have indicated that TSIIA is a natural anti-tumor agent, however, the underlying mechanisms remain largely unknown. We studied the anti-tumor efficacy of TSIIA on human colon cancer cell lines in vitro and in vivo, and have further demonstrated that the molecular anti-cancer property of TanIIA is dependent on its inhibition of proliferation and induction of apoptotic cell death via p38MAPK signaling pathway. The results indicated that TanIIA were markedly more effective than control group in preventing tumor growth and increasing survival time. This phenomenon was accompanied by a decrease in cell growth in a concentration- and time-dependent manner. Hoechst 33258 stainning showed an increased number of apoptotic bodies in the cells after treatment with TanIIA. Flow cytometry analysis showed TanIIA could cause an obvious colon cacer cell apoptosis and arrest cells in the G0/G1 phase. After p38MAPK signaling pathway was blocked, the cell apoptotic rates and cell ratio of G0/G1 phase were decreased significantly. In addition, TanIIA increased p38MAPK phosphorylation, and up-regulated the protein expression of ATF-2, The ATF-2 expression was found to be significantly decreased in cells treated with p38MAPK-specific inhibitor. From our results, it is speculated that TanIIA may induce human colon cancer cells apoptosis and arrest cells in the G0/G1 phase involve up-regulation the expression of the ATF-2 mRNA via p38MAPK signal transduction pathway. Note: This abstract was not presented at the meeting. Citation Format: Yan Wang, Lihong Zhou, Qing Ji, Hua Sui, Xueqing Hu, Gang Cai, Qi Li. TanshinoneIIA, an isolated compound from Salvia miltiorrhiza, inhibits cell growth in colorectal cancer via activation of p38MAPK signaling pathway [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 4303.

MiR-657/ATF2 Signaling Pathway Has a Critical Role in Spatholobus suberectus Dunn Extract-Induced Apoptosis in U266 and U937 Cells.

Though Spatholobus suberectus Dunn (SSD) has been reported to have anti-virus, anti-osteoclastogenesis, and anti-inflammation activities, its underlying anti-cancer mechanism has never been elucidated in association with the role of miR-657 in endoplasmic reticulum (ER) stress-related apoptosis to date. SSD treatment exerted cytotoxicity in U266 and U937 cells in a dose-dependent manner. Also, apoptosis-related proteins such as PARP, procaspase-3, and Bax were regulated by SSD treatment. Furthermore, Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay revealed that a number of apoptotic bodies were increased by SSD. Interestingly, the ER stress-related proteins such as p-ATF2 and CHOP were elevated by SSD. Interestingly, reactive oxygen species (ROS) generation and cytotoxicity by SSD treatment were significantly reduced by N-Acetyl-L-cysteine (NAC). Among the microRNAs (miRNAs) regulated by SSD treatment, miR-657 was most significantly reduced by SSD treatment. However, an miR-657 mimic reversed SSD-induced apoptosis by the attenuation of the expression of p-ATF2, CHOP, and PARP cleavage. Overall, these findings provide scientific evidence that miR657 is an onco-miRNA targeting the ER stress signal pathway and SSD induces apoptosis via the inhibition of miR-657, ROS, and the activation of p-ATF2 and CHOP as a potent anti-cancer agent for myeloid-originated hematological cancer.

Apoptosis in trophoblast in pre-eclampsia and intrauterine growth retardation: A light microscopy study

Background: Pre-eclampsia and Intrauterine growth retardation (IUGR) affects 3% and 10% of all pregnancies. Apoptosis, a programmed cell death is known to occur as a part of both physiological and pathological process. This study was done to calculate apoptotic index in placental trophoblasts of pre-eclampsia, IUGR and compare it with normal term placenta. The aim of this study is to calculate apoptotic index in cytotrophoblast and syncytiotrophoblast in preeclampsia and IUGR and compare it with placenta from normal pregnancyMethods: A total of fifty four placentas were included in the study. 18/54 cases were from pre-eclampsia patients, 18/54 cases were from IUGR patients, 18/54 were age matched controls. Hematoxylin and eosin stained sections from representative were screened under oil immersion using Olympus microscope (100X) for apoptotic bodies and apoptotic index was calculated by counting the total number of apoptotic bodies and dividing it by total number of nuclei counted in the trophoblasts multipled by 100. Data was entered into Microsoft excel data sheet and was analyzed using SPSS 22 version software. ANOVA was the test of significance used. P value<0.05 was considered significant. Result: Apoptotic bodies and index were maximum in cytotrophoblast and syncytiocytotrophoblast of preeclampsia as compared with IUGR and normal placentas. Statistically significant difference was observed in apoptotic index in cytotrophoblast and apoptotic index in Syncytiotrophoblast between three groups.Conclusion: Increased number of apoptotic bodies and index were seen in trophoblast of pre-eclampsia cases as compared with IUGR and normal placentas which may be related to onset of maternal inflammatory syndrome.DOI:10.21276/APALM.1693

Ethanol extract from Cnidium monnieri (L.) Cusson induces cell cycle arrest and apoptosis via regulation of the p53‑independent pathway in HepG2 and Hep3B hepatocellular carcinoma cells.

Cnidiummonnieri (L.) Cusson is a frequently used traditional Chinese medicine that treats gynecological diseases and carbuncles. However, the mechanism of action of C. monnieri remains to be fully elucidated. The present study examined the cell cycle arrest and apoptotic effects resulting from ethanol extract of C. monnieri (CME) in HepG2 (wild‑type p53) and Hep3B (p53‑null) hepatocellular carcinoma cells. An MTT assay was used to confirm the anti‑proliferative effect of CME. The cells were stained with Hoechst 33342 or propidium iodide. It was demonstrated that proliferation of HepG2 cells was suppressed by CME. Cell cycle arrest occurred in the G1 phase following treatment with CME and the number of apoptotic bodies was increased. The expression levels of cell cycle‑associated proteins, including protein kinaseB (Akt), glycogen synthase kinase‑3β (GSK‑3β), p53, cyclin E and cyclin‑dependent kinase2 (CDK2) were determined by western blot analysis. The protein levels of phosphorylated (p)‑Akt, p‑GSK‑3β, p‑MDM2 and cyclinE were decreased, whereas the protein levels of p53, p21 and p‑CDK2 (Thr14/Tyr15) were increased following treatment with CME. Furthermore, treatment or co‑treatment with LY294002 (phosphoinositide‑3‑kinase/Akt inhibitor) or Pifithrin‑α (p53 inhibitor) with CME resulted in CME‑induced G1 arrest which occurred through the p53‑independent signaling pathway in hepatocellular carcinoma cells. In conclusion, CME induces G1 arrest and apoptosis via the Akt/GSK‑3β signaling pathway which is regulated by MDM2‑induced degradation of p21, rather than p53.

Evaluation of apoptosis in skin biopsies of patients of borderline leprosy and lepra type 1 reaction.

Background:The role of apoptosis is not clear in leprosy and lepra reactions.Objectives:To evaluate frequency of apoptosis in skin lesions of borderline leprosy and Type 1 lepra reaction.Methods:Sixty patients with borderline leprosy (30 with clinically diagnosed Type 1 reaction (T1R) (Group I) and 30 without clinical evidence of reaction (Group II)) were analyzed in this prospective study. Apoptosis was detected by two different methods for comparison, that is, histopathologic examination (HPE) and deoxyribonucleic acid (DNA) fragmentation and electrophoresis. Quantification of apoptotic bodies/10 high power fields (HPF) was also done.Results:Out of 30 cases, apoptosis was detected in 29 cases in Group I and 24 cases in Group II by HPE (P = 0.103), whereas, with the use of DNA electrophoresis it was detected in 24 cases in Group I and 18 cases in Group II (P = 0.091). On quantitative estimation it was found that number of apoptotic bodies are higher in Group I in comparison to Group II (2.77 vs 1.99), which is statistically significant.Conclusions:There was moderate agreement (κ = 0.47) between the two methods of apoptosis detection. Apoptosis was seen more in patients with T1R both qualitatively (statistically nonsignificant) and quantitatively (statistically significant). Clinical significance of this novel finding is that apoptosis can be used as one of the variables for diagnosis of T1R to increase detection rate.