The cerebellum is known to control balance, equilibrium, and muscle tone. If the cerebellum becomes damaged, the body is unable to retain its balancing functions or involuntary muscle movement. This is why, in stroke patients, there is a high risk of functional disability, as well as a myriad of other disabilities secondary to stroke. Ischemia was induced in SD mice by occluding the common carotid artery for 5 minutes, after which blood was reperfused. Needle electrode electrical stimulation(NEES) was applied to acupuncture points, at 12, 24, and 48 hours post-ischemia on the joksamri. Protein expression was investigated through caspase-3 antibody immuno-reactive cells in the cerebral nerve cells and Western blotting. The results were as follows: The number of caspase-3 reactive cells in the corpus cerebellum 12 and 24 hours post-ischemia was significantly (p<.05) smaller in the NEES group compared to the GI group. caspase-3 expression 12 and 24 hours post-ischemia was significantly(p<.05) smaller in the NEES group compared to the GI group. Based on these results, NEES seems to have a significant effect on Caspase-3 in the cerebellum in an ischemic state at 12 and 24 hours post ischemia, NEES delays the occurrence of early stage apoptosis-inducing Caspase-3, delaying and inhibiting apoptosis. Further systematic studies will have to be conducted in relation to the application of this study’s results on stroke patients.
Adult stem cell transplantation has been increased every year, because of the lack of organ donors for regenerative medicine. Therefore, development of reliable and safety cryopreservation and bio-baking method for stem cell therapy is urgently needed. The present study investigated safety of dimethyl sulfoxide (DMSO) such as common cryoprotectant on porcine bone marrow derived mesenchymal stem cells (pBM-MSCs) by evaluating the activation of Caspase-3 and -7, apoptosis related important signal pathway. pBM-MSCs used for the present study were isolated density gradient method by Ficoll-Paque Plus and cultured in A-DMEM supplemented 10% FBS at in 5% incubator. pBM-MSCs were cryopreserved in A-DMEM supplemented either with 5%, 10% or 20% DMSO by cooling rate at /min in a Kryo 360 (planner 300, Middlesex, UK) and kept into . Survival rate of cells after thawing did not differ between 5% and 10% DMSO but was lowest in 20% DMSO by 0.4% trypan blue exclusion. Activation of Caspase-3 and -7 by Vybrant FAM Caspase-3 and -7 Assay Assay Kit (Molecular probes, Inc.OR, USA) was analyzed with a flow cytometer. Both of cryopreserved and control groups (fresh pBM-MSCs) were observed after the activation of Caspase-3 and -7. The activation did not differ between 5% and 10% DMSO, but was observed highest in 20% DMSO. Therefore 5% DMSO can be possibly used for cell cryopreservation instead of 10% DMSO.
당뇨망막병증은 서구에서 성인들의 실명을 일으키는 원인이다. 당뇨병이 있을 때 고혈당증은 여러 세포 형태에서 세포자연사를 유도하지만 그 기작은 명확하게 밝혀지지 않았다. 본 연구의 목적은 인간망막 내피세포에서 고혈당 포도당이 세포자연사에 미치는 영향에 대하여 알아보았다. 망막 내피세포는 5, 25, 50 mM 포도당이 포함된 IMDM배지에서 37℃, 5% CO2조정된 항온기에서 24, 36, 48시간 동안 배양하였다. 여러 농도의
Cordyceps militarisis well known as a traditional herbal ingredient, which has been used for patients suffering from cancer in oriental medicine. In this study we have investigated the biochemical mechanisms of anti-proliferative effects by C. militarisextract(CME) in human breast cancer MDA-MB-231 cells. It was found that CME treatment induced chromatin condensation, mitochondrial energization, annexin V staining and sub-G1 phase DNAcontent. These indicators of apoptosis correlate with the mitochondrial dependent pathway, which results in the activation of caspase-3 activity. Both the cytotoxic effect by CME treatment were significantly inhibited by z-DEVD-fmk, a caspase-3 inhibitor,demonstrating the important role of caspase-3 in the observed cytotoxic effect. Cotreatment of CME and LY294002, resulted in significantly induction of apoptosis. These results indicate that caspase-3 is a key regulator of apoptosis in response to CME in human breast cancer MDAMB- 231 through downregulation of Akt, and that the C. militaris extract may therefore have therapeutic potential against human breast cancer.
Cordyceps militaris is well known as a traditional herbal ingredient, which has been used for patients suffering from cancer in oriental medicine. In this study we have investigated the biochemical mechanisms of anti-proliferative effects by C. militaris extract(CBE) in human breast cancer MDA-MB-231 cells. It was found that CBE treatment induced chromatin condensation, mitochondrial energization, annexin V staining and sub-G1 phase DNA content. These indicators of apoptosis correlate with the mitochondrial dependent pathway, which results in the activation of caspase-3 activity. Both the cytotoxic effect by CBE treatment were significantly inhibited by z-DEVD-fmk, a caspase-3 inhibitor, demonstrating the important role of caspase-3 in the observed cytotoxic effect. Co-treatment of CBE and LY294002, resulted in significantly induction of apoptosis. These results indicate that caspase-3 is a key regulator of apoptosis in response to CBE in human breast cancer MDA-MB-231 through down regulation of Akt, and that the C. militaris extract may therefore have therapeutic potential against human breast cancer.
Cordyceps militaris is well known as a traditional herbal ingredient, which has been used for patients suffering from cancer in oriental medicine. In this study we have investigated the biochemical mechanisms of anti-proliferative effects by C. militaris extract(CBE) in human breast cancer MDA-MB-231 cells. It was found that CBE treatment induced chromatin condensation, mitochondrial energization, annexin V staining and sub-G1 phase DNA content. These indicators of apoptosis correlate with the mitochondrial dependent pathway, which results in the activation of caspase-3 activity. Both the cytotoxic effect by CBE treatment were significantly inhibited by z-DEVD-fmk, a caspase-3 inhibitor, demonstrating the important role of caspase-3 in the observed cytotoxic effect. Co-treatment of CBE and LY294002, resulted in significantly induction of apoptosis. These results indicate that caspase-3 is a key regulator of apoptosis in response to CBE in human breast cancer MDA-MB-231 through down regulation of Akt, and that the C. militaris extract may therefore have therapeutic potential against human breast cancer.
Agaricus blazeiMurill is an edible mushroom distributed in Brazil and presently cultivated in other areas, including Korea, Japan, and China. Its chemical components, including steroids and lectin and various polysaccharides have been widely studied. For this, we used U937/vector and U937/Bcl-2 cells, which were generated by transfection of the cDNA of the Bcl-2 gene. As compared with U937/vector, U937/Bcl-2 cells exhibited a 4-fold greater expression of Bcl-2. Treatment with 0.5 or 4 mg/ml A. blazei Murill for 24 h produced morphological features of apoptosis in U937/vector cells, respectively. This was associated with caspase-3 activation and PARP degradation. In contrast, A. blazei Murill-induced caspase-3 activation and PARP degradation and apoptosis were significantly inhibited by z-DEVD-fmk in U937 cells. Bcl-2 overexpressing cells exhibited sustained caspase-3 activation and expression levels of the Bcl-2 proteins during A. blazei Murill-induced apoptosis. In addition, these findings indicate that Bcl-2 inhibits A. blazei Murill-induced apoptosis by a mechanism that interferes with Bcl-2 degradation and activity of caspase-3 that is involved in the execution of apoptosi.
In most tissues, apoptosis plays a pivotal role in normal development and in regulation of cell number. Therefore inappropriate apoptosis is revealed in a variety of diseases. This study was carried out to investigate the effects of acupuncture and needle electrode electrical stimulation on the change of caspase-3, 9 and neuronal nitric oxide synthase (nNOS) immunoreactive cells in the sprague dawley rats (SD rat). In immobilized SD rats (n=5), enhanced caspase-3 and caspase-9 expression were detected in the reticular part of substantia nigra, and enhanced nNOS was detected in the dorsolateral periaqueductal gray (DL-PAG) of midbrain and the paraventricular nucleus (PVN) of the hypothalamus using immunohistochemistry. Following the immobilization, acupuncture (n=5) and needle electrode electrical stimulation (n=5, 2 Hz) was applied at Hg (LI4) acupoint of SD rats, respectively. The stress-induced enhancement in the expression of caspase-3, 9 and nNOS were The present results demonstrate that and needle electrode electrical stimulation are effective in the modulation of expression of caspase-3, 9 and nNOS induced by immobilization.
Tributyltin (TBT) used world-wide in antifouling paints toy ships is a wide-spread environmental pollutant. At low doses, antiproliferative modes of action have been shown to be involved, whereas at higher doses apoptosis seems to be the mechanism of toxi
포유류 난포의 폐쇄 과정은 매우 정교한 내분비적 조절작용에 의해 일어나며, 이 과정중에 발생하는 난포 내 과립세포의 퇴화는 핵응축 현상을 동반하는 것으로 알려져 있다. 본 연구는 핵응축 현상과 관련하여 돼지 난소 내 폐쇄난포의 과립세포가 퇴화 시 동반되는 세포 사멸이 아포토시스의 과정에 의해서 일어나는지의 여부와 아포토시스 관련 주요 단백질 분해 효소인 캐스파제-3과 연관된 세포 사멸 기전과 관련이 있는지에 대해서 조사하고자 하였다. 돼지 난소로부터 정