Methylglyoxal is a highly reactive precursor which forms advanced glycation end products (AGEs). AGEs and methylglyoxal are known to induce various diseases such as diabetes, vascular disorders, Diabetes Mellitus (DM), and neuronal disorders. Juglans regia L is an important food commonly used worldwide, having nutritious components, including phenolic compounds. Since ancient times, Juglans regia L have been differently applied by various countries for health and in diverse diseases, including arthritis, asthma, skin disorders, cancer, and diabetes mellitus. However, the effect of diabetes-induced renal damage against AGEs remains unclear. This study evaluates the anti-glycation and renal protective effects of ethanol extract of Juglans regia L against methylglyoxal-induced renal tubular epithelial cell death. Exposure to methylglyoxal resulted in reduced cell viability in NRK-52E cells, but co-treatment with Juglans regia L extracts significantly increased the cell viability. In addition, we examined the anti-glycation effect of Juglans regia L extracts. Compared to the positive control aminoguanidine and Alagebrium, treatment with Juglans regia L extracts significantly inhibited the formation of AGEs, collagen cross-linking, and breaking collagen cross-linking. Taken together, our results indicate that Juglans regia L is a potential therapeutic agent for regulating diabetic complications by exerting anti-glycation and renal protective activities.

Bovine mammary epithelial (MAC-T) cells are commonly used to study mammary gland development and mastitis. Lipopolysaccharide is a major bacterial cell membrane component that can induce inflammation. Autophagy is an important regulatory mechanism participating in the elimination of invading pathogens. In this study, we evaluated the mechanism underlying bacterial mastitis and mammary cell death following lipopolysaccharide treatment. After 24 h of 50 μg/mL lipopolysaccharide treatment, a significant decrease in the proliferation rate of MAC-T cells was observed. However, no changes were observed upon treatment of MAC-T cells with 10 μg/mL of lipopolysaccharide for up to 48 h. Thus, upon lipopolysaccharide treatment, MAC-T cells exhibit dose-dependent effects of growth inhibition at 10 μg/mL and death at 50 μg/mL. Treatment of MAC-T cells with 50 μg/mL lipopolysaccharide also induced the expression of autophagy-related genes ATG3, ATG5, ATG10, ATG12, MAP1LC3B, GABARAP-L2, and BECN1. The autophagy-related LC3A/B protein was also expressed in a dose-dependent manner upon lipopolysaccharide treatment. Based on these results, we suggest that a high dose of bacterial infection induces mammary epithelial cell death related to autophagy signals.

Despite evidence that bacteria-sensing Toll-like receptors (TLRs) are activated in salivary gland tissues of Sjogren syndrome (SS) patients, the role of oral bacteria in SS etiopathogenesis is unclear. We previously reported that two SS-associated oral bacteria, Prevotella melaninogenica (Pm) and Rothia mucilagenosa (Rm), oppositely regulate the expression of major histocompatibility complex class I (MHC I) in human salivary gland (HSG) cells. Here, we elucidated the mechanisms underlying the differential regulation of MHC I expression by these bacteria. The ability of Pm and Rm to activate TLR2, TLR4, and TLR9 was examined using TLR reporter cells. HSG cells were stimulated by the TLR ligands, Pm, and Rm. The levels of MHC I expression, bacterial invasion, and viability of HSG cells were examined by flow cytometry. The hypoxic status of HSG cells was examined using Hypoxia Green. HSG cells upregulated MHC I expression in response to TLR2, TLR4, and TLR9 activation. Both Pm and Rm activated TLR2 and TLR9 but not TLR4. Rm-induced downregulation of MHC I strongly correlated with bacterial invasion and cell death. Rm-induced cell death was not rescued by inhibitors of the diverse cell death pathways but was associated with hypoxia. In conclusion, Pm upregulated MHC I likely through TLR2 and TLR9 activation, while Rm-induced hypoxia-associated cell death and the downregulation of MHC I, despite its ability to activate TLR2 and TLR9. These findings may provide new insight into how oral dysbiosis can contribute to salivary gland tissue damage in SS.

The role of transient receptor potential vanilloid receptor-1 (TRPV1) has been primarily investigated in pain sensory neurons. Relatively, little research has been performed in testicular cells. TRPV1 is abundantly expressed in Leydig cells of young adult mice. This study was conducted to determine the role of the TRPV1 channel in Leydig cells. TRPV1 modulators and testosterone were treated to the mouse Leydig cell line TM3 cells for 24 h. Capsaicin, a TRPV1 activator, dose-dependently induced cell death, whereas capsazepine, a TRPV1 inhibitor, inhibited capsaicin-induced cell death. Testosterone treatment reduced capsaicin-induced cell death. High concentrations of testosterone decreased TRPV1 mRNA and protein expression levels. However, TRPV1 modulators did not affect testosterone production. These results showed that capsaicin induced cell death of Leydig cells and that testosterone reduced capsaicininduced cell death. Our findings suggest that testosterone may regulate the survival of Leydig cells in young adult mice by decreasing the expression level of TRPV1.

In this study, to analyze whether artificial regulation of apoptosis in the development of somatic cells can affect the stable growth and development of cells, 20 alpha-hydroxysteroid dehydrogenase (20α-HSD) and rapamycin were treated to induce apoptosis and autophagy in the both skin and muscle cells. Respectively, and 3-methyladenine was supplemented to inhibit cell death. Our results show that stimulation with rapamycin activated autophagy in both types of cells, but increased apoptosis more than autophagy in the case of skin cells. These results indicate that there was a difference in the expression of survival factors and cell development depending on the type of cell. In particular, in the expression of autophagy-related gene (MAP1LC3A) was higher than that of Casp-3, an apoptosis factor. Furthermore, cell development was the highest in all cell groups cultured by artificially inducing autophagy, however the lowest in the apoptosis-inhibited group. Especially, the noteworthy result of this study was that when apoptosis was induced using 20α-HSD, it was possible to induce apoptosis in both skin and muscle cells. Therefore, the main point of this study is that apoptosis induced during cell culture plays a pivotal role in cell remodeling.

Cancer is the second leading cause of death worldwide and currently there are many approaches developing towards cancer treatment. Cancer treatments like chemotherapy and radiation therapy are often painful and have adverse effects. The mechanism of apoptosis is a complex process and it involves different pathways in its mechanism of action. Apoptosis can be caused by signals within the cell such as stress, or by extrinsic signals such as ligands binding to cell surface death receptors. The programmed cell death plays a important role in the several physiological and pathological processes. It plays important role in homeostasis. Flavonoids have gained importance as anticancer agents promoting cytotoxicity and apoptosis in cancer cells. Flavonoids are present in many medicinal plants which are a kind of ubiquitous natural products and essential active ingredients. They have strong biological activities with high efficiency and low toxicity, possessing good preventive and cure effects on different tumor forms. Flavonoids such as Scutellarein, Pectolinarigenin and Naringin have reported to possess significant anti-cancer effects on different cancer cell lines till date. In this review, we provide a summary about the anti-cancer effect of the three flavonoids and its mechanisms of action that can be used in future for understanding their potent anti-tumor properties.

Cognitive impairment is considered to be key research topics in the field of neurodegenerative diseases and in understanding of learning and memory. In the present study, we investigated neuroprotective effects of Schisandra chinensis (SC) and Ribes fasciculatum (RF) extracts in hydrogen peroxide-induced neuronal cell death in vitro and scopolamine-induced cognitive impairment in Sprague Dawley® (SD) rat in vivo. Apoptotic cell death in neuroblastic PC12 cell line was induced by hydrogen peroxide for 1 hour at 100 μM. However, mixture of SC and RF treatment prevented peroxide induced PC12 cell death with no neurotoxic effects. For in vivo experiment, the effect of SC and RF extracts on scopolamine-induced cognitive impairment in SD rat was evaluated by spontaneous alternation behavior in Y-Maze test. After 30 min scopolamine injection, the scopolamine-induced rats presented significantly decreased % spontaneous alteration and acetylcholine level, compared to non-induced group. However, treatment of SC+RF extracts rescued the reduced % spontaneous alteration with acetylcholine concentration from hippocampus in scopolamineinduced rats. These results suggested that mixture of SC and RF extract may be a potential natural therapeutic agent for the prevention of cognitive impairment.

The purpose of this study was to investigate diethylnitrosamine (DEN)-induced liver damage in zebrafish. Zebrafish larvae were divided into five groups after seventy-two hours fertilization: group 1 (G1) as control, group 2 (G2) as probe control, groups 3, 4, and 5 (G3, G4, and G5) as DEN treated at doses of 25, 50, and 100 μg/mL, respectively. At twenty-two hours after DEN treatment, groups 2, 3, 4, and 5 were treated with ApoFlamma H 675 at a dose of 100 μM/zebrafish. They were examined by fluorescence stereomicroscope at twenty-four hours after DEN treatment. After fixation, the zebrafish were processed, embedded, sectioned and stained with hematoxylin and eosin (HE) and terminal deoxynucleotide transferase dUTP nick end labeling (TUNEL) staining. Fluorescence intensity of the livers of G3, G4, and G5 was significantly increased compared with those of G1 (p<0.01). Furthermore, fluorescence intensity of the livers of G3 and G5 was significantly increased compared with those of G2 (p<0.05 and p<0.01). HE staining showed cell deaths in the livers of DEN-treated zebrafish and TUNEL staining confirmed cell death in the same location. Taken together, in vivo fluorescence bioimaging detected cell death in the liver of DEN-treated zebrafish. This outcome was confirmed with histopathological examination. The results of this study provide confidence for using zebrafish as a liver carcinogenesis model.

In the present study of this experiment was to understand the expression of apoptotic gene expression in the ovary of miniature pigs and pigs on the 15th day of estrus. Also the compare and analyze of programmed cell death type(Apoptosis and autophagy) expression pattern during mature oocyte on the miniature and normal pig cells. Analysis of mRNA gene expression of ovary in miniature and normal pigs on the 15th day of estrus showed that the expression of genes related to Autophagy (ATG13, MAP1LC3, Beclin1) was high in normal pigs but the expression of ATG1 and ATG5 genes was low. In addition, the expression of genes related to apoptosis (Casp-3, BAX) was high in the mini pigs, and the gene related to the LH hormone was high in the miniature pigs, whereas the expression of the gene related to the FSH hormone was high in the normal pigs. On the other hand, the result of muture oocyte on the miniature and normal pig cells is the expression of Casp-3 protein was moust high from treatment of FL+rapa (FSH+LH and Rapamycin) of the oocyte on the miniature pig cell. However, MAP1LC3 expression was higher in the oocytes of treatment of rapanycine treatment on the nomal pig cells. There was no gene expression in cumulus cells of matured oocytes in mini pig cells, whereas MAP1LC3 expression was higher in oocyte cumulus cells matured in normal pig cells. It was confirmed that the miniature and normal pigs showed different programmed cell death patterns, In the case of oocytes matured in miniature pig cells, MAP1LC3 gene expression was found to be low in spite of treatment with Autophagy regulator.

알츠하이머병의 발병원인과 기전에 대하여 많은 연구가 진행되었으나, 아직까지 완전히 밝혀지지 않고 있다. 이에 본 연구에서는 STZ로 유도된 세포독성으로부터 노인성 질병의 예방과 항산화 효과로 잘 알려진 비타민 C를 이용하여 SH-SY5Y 신경세포 내 보호 기전을 살펴보고자 하였다. 본 연구 결과에 의하면, STZ를 SH-SY5Y 신경세포에 처리하였을 때, 세포사멸이 유도되는 것을 확인하였으며, 비타민C를 전처리함으로써 세포생존도가 증가하는 것을 확인하였다. 비타민 C가 신경세포를 보호하는 기전을 알아보기 위하여 세포자멸사 과정에서 신호전달을 통해 다양한 유전자를 발현시키는 MAPKs의 인산화를 살펴보았다. 비타민 C 처리시 ERK의 인산화가 증가하며, 세포증식을 유도하는 것으로 확인하였으며, ERK와는 반대로 염증, 세포사멸과 관련된 JNK의 인산화는 비타민 C에 의해 인산화가 억제되는 것으로 확인되었다. 또한 STZ는 Bax 단백질의 발현 증가와 Bcl-2 단백질의 발현을 감소시켰으며, apoptosis antibody array로 확인한 결과, Cytochrome C가 유도됨을 확인하였다. 이뿐만 아니라, 비타민 C 처리에 의해 Bcl-2가 증가하여 세포사멸이 억제되는 것을 확인하였으며, 항산화 효소인 Sod-1의 발현을 증가하는 것 또한 확인하였다. 이는 비타민 C가 항산화 방어체계를 강화시켜 STZ에 의한 세포손상을 보호하는 것으로 사료된다. 본 연구를 통하여 STZ로 유도된 SH-SY5Y 신경세포 손상 에서 비타민 C가 여러 기전을 통하여 세포자멸사를 억제하여 세포를 보호하는 효과가 있는 것을 확인할 수 있었다. 그러나 연구는 일부 기전들만을 확인한 것으로 추후 연구에서는 다양한 비타민 C의 농도 및 처리시간에 따른 기전의 차이를 분석하고, 실험동물을 통한 검증 및 스트렙토조토신에 의한 아밀로이드베타와 타우의 관련성을 확인하고자 한다.

Actinidia arguta (Actinidiaceae), which is commonly referred to as hardy kiwifruit, has been reported to possess anti-inflammatory, anti-allergic and antioxidative properties. The protective effect of the leaves and stems of A. arguta against amyloid β protein (Aβ) (25-35)-induced cultured neuronal cell death and memory impairment was investigated in the current study. Exposure of cultured cortical neurons to 10 μM Aβ (25-35) for 24 h induced significant neuronal death as assessed by a 3-[4,5-dimethylthiazol- 2-yl]-2,5-diphenyl-tetrazolium bromide (MTT) assay and Hoechst 33342 staining. However, A. arguta (10 and 50 μg/ ml) prevented Aβ (25-35)-induced apoptotic neuronal death in cultured cortical neurons. A. arguta also inhibited the 100 μM H2O2-induced decrease of the MTT reduction rate in cultured neurons. Memory impairment was produced by intracerebroventricular microinjection of 15 nmol Aβ (25- 35) and examined using the passive avoidance test in ICR mice. Chronic treatments with A. arguta (50 and 100 mg/ kg, 14 days, p.o.) significantly prevented memory impairment induced by Aβ (25-35), and A. arguta inhibited the Aβ (25-35)-induced increase of cholinesterase activity in the brains of memory impaired mice. These results suggest that A. arguta might be able to inhibit Aβ (25-35)-induced neuronal death and memory impairment via antioxidative and anti-cholinesterase effects and that A. arguta could have a therapeutic role for preventing the progression of neurodegeneration in Alzheimer’s disease.

Amyloid-β protein (Aβ) is known to increase free radical production in neuronal cells, leading to cell death by oxidative stress. The purpose of this study was to evaluate the protective effects of PineXol® on Aβ25-35 induced neuronal cell death. Rat pheochromocytoma (PC-12) cells were pre-treated with 100 μg/mL of PineXol® for 2 h. The cells were exposed to single dose of 30 μM Aβ25-35 for 24 h. Cell death was assessed by a cell count kit-8 (CCK-8) assay, lactate and dehydrogenase (LDH) release assay. An Apoptotic process was analyzed by a protein expression of the Bcl-2 family using western blotting. Cell viability increased in PC-12 cells treated with both Aβ25-35 and PineXol®, compared to the control group. PineXol® induced a decrease of the Bcl-2 protein expression (p<0.05), while Bax and Sod1 increased (p<0.05), indicating attenuation of Aβ25-35 induced apoptosis. These results suggest that PineXol® may be a good candidate for the prevention of Alzheimer’s disease(AD).

The nature of molecular mechanisms governing embryo development is largely unknown, but recent reports have demonstrated that proper execution of programmed cell death is crucial for this process. The main objective of this study is to examine the mode of programmed cell death during nuclear transfer embryos development in porcine. In particular, the relative employment of two major pathways in programmed cell death; e.g. apoptosis (type I) and autophagy (type II) was compared. Oocytes use in the study was matured in vitro in the presence of 10% FBS maturation medium. After nuclear transfer embryos were cultured for each programmed cell death control factor [Cysteamine(Cyst : 0.4mM), 3-methyladenine(3MA : 2.5mM) and Rapamycin(RP : 100nM)] in TCM-199 medium supplemented with 0.1% BSA. In this study results of among the blastocysts development in 3MA; PCNA, MAP1LC3A and ATG5 RNA gene expression level increased in the order of IVF<Cyst < 3MA < RP. However Casp-3 and TNF-r RNA gene expression level decreased in the order of IVF < 3MA and RP< Cyst. The expression of mTOR showed a pattern opposite to that of MAP1LC3A. That is, its expression was the lowest in Cyst group. And next experiments analysis of MMP expression patterns. Analysed this MMPs enzyme activation to evaluate the effectiveness of high quality brastocyst culture in porcine. In this results of the enzymatic activity of MMP-2 and MMP-9 was assessed in culture, the level of active MMP-9 was higher expression in the medium of each 3MA and RP treatment group, with the level of another treatment group being relatively higher. These results suggest that the autophagy activation culture medium is more effective for stable and innovative nuclear transfer embryos development.

Background: Periodontitis is generally a chronic disorder characterized by the breakdown of tooth-supporting tissues. P. gingivalis, a Gram-negative anaerobic rod, is one of the major pathogens associated with periodontitis. Frequently, P. gingivalis infection leads to cell death. However, the correlation between P. gingivalis–induced cell death and periodontal inflammation remains to be elucidated. Among cell deaths, the death of immune cells appears to play a significant role in inflammatory response. Thus, the aim of this study was to examine P. gingivalis–induced cell death, focusing on autophagy and apoptosis in THP-1 cells. Methods: Human acute monocytic leukemia cell line (THP-1) was used for all experiments. Autophagy induced by P. gingivalis in THP-1 cells was examined by Cyto ID staining. Intracellular autophagic vacuoles were observed by fluorescence microscopy using staining Acridine orange (AO); and 3-methyladenine (3-MA) was used to inhibit autophagy. Total cell death was measured by LDH assay. Cytokine production was measured by an ELISA method. Results: P. gingivalis induced autophagy in an MOI-dependent manner in THP-1 cells, but 3-MA treatment decreased autophagy and increased the apoptotic blebs. P. gingivalis infection did not increase apoptosis compared to the control cells, whereas inhibition of autophagy by 3-MA significantly increased apoptosis in P. gingivalis-infected THP-1 cells. Inhibition of autophagy by 3-MA also increased total cell deaths and inflammatory cytokine production, including IL-1β and TNF-⍺. Conclusion: P. gingivalis induced autophagy in THP-1 cells, but the inhibition of autophagy by 3-MA stimulated apoptosis, leading to increased cell deaths and pro-inflammatory cytokines production. Hence, the modulation of cell deaths may provide a mechanism to fight against invading microorganisms in host cells and could be a promising way to control inflammation.

Taxol is an anti-cancer agent that stabilizes the microtubules of cancer cells, resulting in inhibition of mitosis and thus preventing the proliferation of cancer cells. However, many anti-cancer agents including taxol work on normal cells as well as cancer cells, resulting in side effects such as immunosuppression. A marine algae-derived sulfated polysaccharide, fucoidan, an anti-cancer agent, also showed immunostimulating effects. This study investigated the effects of fucoidan on taxol-treated spleen cells. Spleen cells were treated with taxol in a concentration-dependent manner and in combination with fucoidan. MTT assay and flow cytometry analysis were performed to measure the viability and activity of treated cells. Two assays demonstrated that taxol induced the death of spleen cells. Fucoidan clearly inhibited the cell death induced by taxol. In addition, fucoidan enhanced the production of nitric oxide in spleen cells, which was decreased by taxol. Taken together, taxol can induce the cell death of spleen cells, a major type of immune cells and fucoidan protects spleen cells from taxol-induced cell death. This finding suggests that taxol and fucoidan can be used in combination for lowering the immunosuppressive effects of taxol.

The nature of molecular mechanisms governing embryonic cell block is largely unknown, but recent reports have demonstrated that proper execution of programmed cell death is crucial for this process. The main objective of this study is to determine effects of programmed cell death on porcine oocytes development in vitro after parthenogenesis. Among the blastocysts matured in 3MA, MAP1LC3A and ATG5 RNA gene expression level increased in the order of Cyst < 3MA < RP. However, Casp-3 and TNF-r RNA gene expression level decreased in the order of RP < 3MA < Cyst. Expression of mTOR within the RP-cultured blastocyst was the most highly to the inner cell mass, while 3MA-cultured blastocyst showed very lowest expression in inner cell mass. The expression of mTOR showed a pattern opposite to that of MAP1LC3A. That is, its expression was the lowest in Cyst group. When the enzymatic activity of MMP-2 and MMP-9 was assessed in culture, the level of active MMP-9 was higher expression in the medium of each RP treatment group, with the level of another treatment group being relatively higher. Analyses of TIMP-2 and TIMP-3 revealed that their expression was higher in groups that did not receive RP treatment. More specifically, the level of TIMP-2 was not affected by Cyst treatment, while the level of TIMP-3 was higher in 3MA and RP treatment group. There was highly cell division activation efficiency of parthenogenesis on cultured system of RP supplement IVC medium. Therefore, these results suggest that embryo development was significantly increased in conditional culture medium with active autophagy as compared to common cultured condition. Further investigation of this distinction may enable the development of innovative improvements for the production of porcine somatic cell nuclear transfer.

Autophagy is conserved response to starvation by which cells catabolize their components to create an internal supply of essential nutrients. Ceramide is known to induce autophagy in many cells through down-regulation of amino acid and glucose transporters. The mechanism of starvation induced-autophagy in mouse embryo remains unclear. In order to understand the mechanism by which starvation regulates autophagy, in this study, we investigated nutrient transporters expression and the effect of c2-ceramide on the in vitro development, apoptosis and autophagy via starvation in mouse embryo. Glucose transporters (Glut1 and Glut 3), high levels of transcript were expressed from 1 to 2 cells and gradually decreased through the morula and blastocyst (BL) stages. Amino acid transporters (LAT-1 and 4F2hc) gradually decreased from the zygote to the BL stage. Furthermore, the expression of nutrient transporters (Glut1, 3, LAT-1 and 4F2hc) were significantly reduced at the BL stage after ceramide treatment. Especially, mTOR expression after ceramide treatment of embryos was significantly higher than controls. Ceramide treated embryos exhibited significantly reduced developmental rates and total cell numbers, and increased apoptotic cell death at the BL stage. Consequently, we next evaluated the effect of ceramide treatment on mitochondrial number and morphology. There was a significant decrease in the average mtDNA copy number and the mitochondrial area in ceramide treated BL stage embryos. Both the expression of autophagy-related genes, Lc3, Gabarap, Atg4A and Atg4B, and the synthesis of LC3 were significantly induced at the BL stage. These results suggest that autophagy under starvation condition influences the in vitro development and apoptosis and autophagy, and may play a role in early mouse embryogenesis.