• Animal cells and systems
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• 제5권4호
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• pp.267-274
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• 2001

Calpains are a family of cysteine proteases existing primarily in two forms designated by the $Ca^{2+}$ concentration needed for activation in vitro, $\mu$-calpain (calpain-I) and m-calpain (calpain-II). The physiologica1 roles of calpains remain unclear. Many groups have proposed a role for calpains In apoptosis, but their patterns of activation are not well characterized. Calpains have been implicated in neutrophil apoptosis, glucocorticoid-induced thymocyte apoptosis, as well as many other apoptotic pathways. Calpain activation in apoptosis is usually linked upstream or downstream to caspase activation, or in a parallel pathway alongside caspase activation. Calpains have been suggested to be involved in DNA fragmentation (via endonuclease activation), but also as effector proteases that cleave cellular proteins involved in DNA repair, membrane associated proteins and other homeostatic regulatory proteins. Recently, our laboratory demonstrated $\mu$-calpain activation in NAD(P)H: quinone oxidoreducatse 1 (NQO1)-expressing cells after exposure to $\beta$-lapachone, a novel quinone and potential chemo- and radio-therapeutic agent. Increased cytosolic $Ca^{2+}$ in NQO1-expressing cells after $\beta$-lapachone exposures were shown to lead to $\mu$-calpain activation. In turn, $\mu$-calpain activation was important for substrate proteolysis and DNA fragmentation associated with apoptosis. Upon activation, $\mu$-calpain translocated to the nucleus where it could proteolytically cleave PARP and p53. We provided evidence that $\beta$-lapachone-induced, $\mu$-calpain stimulated, apoptosis did not involve any of the known caspases; known apoptotic caspases were not activated after $\beta$-lapachone treatment of NQO1-expressing cells, nor did caspase inhibitors have any effect on $\beta$-1apachone-induced cell death. Elucidation of processes by which $\beta$-1apachone-stimulated $\mu$-calpain activation and calpains ability to activate endonucleases and induce apoptosis independent of caspase activity will be needed to further develop/modulate $\beta$-lapachone for treatment of human cancers that over-express NQO1.

• Journal of Ginseng Research
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• 제47권1호
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• pp.144-154
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• 2023

Background: As the major pathophysiological feature of obstructive sleep apnea (OSA), chronic intermittent hypoxia (CIH) is vital for the occurrence of cardiovascular complications. The activation of calpain-1 mediates the production of endothelial reactive oxygen species (ROS) and impairs nitric oxide (NO) bioavailability, resulting in vascular endothelial dysfunction (VED). Ginsenoside Rg1 is thought to against endothelial cell dysfunction, but the potential mechanism of CIH-induced VED remains unclear. Methods: C57BL/6 mice and human coronary artery endothelial cells (HCAECs) were exposed to CIH following knockout or overexpression of calpain-1. The effect of ginsenoside Rg1 on VED, oxidative stress, mitochondrial dysfunction, and the expression levels of calpain-1, PP2A and p-eNOS were detected both in vivo and in vitro. Results: CIH promoted VED, oxidative stress and mitochondrial dysfunction accompanied by enhanced levels of calpain-1 and PP2A and reduced levels of p-eNOS in mice and cellular levels. Ginsenoside Rg1, calpain-1 knockout, OKA, NAC and TEMPOL treatment protected against CIH-induced VED, oxidative stress and mitochondrial dysfunction, which is likely concomitant with the downregulated protein expression of calpain-1 and PP2A and the upregulation of p-eNOS in mice and cellular levels. Calpain-1 overexpression increased the expression of PP2A, reduced the level of p-eNOS, and accelerated the occurrence and development of VED, oxidative stress and mitochondrial dysfunction in HCAECs exposed to CIH. Moreover, scavengers of O₂^·-, H₂O₂, complex I or mitoKATP abolished CIH-induced impairment in endothelial-dependent relaxation. Conclusion: Ginsenoside Rg1 may alleviate CIH-induced vascular endothelial dysfunction by suppressing the formation of mitochondrial reactive oxygen species through the calpain-1 pathway.

• Parasites, Hosts and Diseases
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• 제52권5호
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• pp.459-469
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• 2014

Entamoeba histolytica is a tissue-invasive protozoan parasite causing dysentery in humans. During infection of colonic tissues, amoebic trophozoites are able to kill host cells via apoptosis or necrosis, both of which trigger IL-8-mediated acute inflammatory responses. However, the signaling pathways involved in host cell death induced by E. histolytica have not yet been fully defined. In this study, we examined whether calpain plays a role in the cleavage of pro-survival transcription factors during cell death of colonic epithelial cells, induced by live E. histolytica trophozoites. Incubation with amoebic trophozoites induced activation of m-calpain in a time- and dose-dependent manner. Moreover, incubation with amoebae resulted in marked degradation of STAT proteins (STAT3 and STAT5) and NF-${\kappa}B$ (p65) in Caco-2 cells. However, $I{\kappa}B$, an inhibitor of NF-${\kappa}B$, was not cleaved in Caco-2 cells following adherence of E. histolytica. Entamoeba-induced cleavage of STAT proteins and NF-${\kappa}B$ was partially inhibited by pretreatment of cells with a cell-permeable calpain inhibitor, calpeptin. In contrast, E. histolytica did not induce cleavage of caspase-3 in Caco-2 cells. Furthermore, pretreatment of Caco-2 cells with a calpain inhibitor, calpeptin (but not the pan-caspase inhibitor, z-VAD-fmk) or m-calpain siRNA partially reduced Entamoeba-induced DNA fragmentation in Caco-2 cells. These results suggest that calpain plays an important role in E. histolytica-induced degradation of NF-${\kappa}B$ and STATs in colonic epithelial cells, which ultimately accelerates cell death.

• 한국축산식품학회지
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• 제25권3호
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• pp.310-315
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• 2005

본 연구는 한우고기의 품질에 영향을 미치는 사후 대사변화를 구명하고자 사후 1, 3, 9, 24시간별에 따라 한우 거세우 배최장근의 pH, 도체온도, R-value 및 단백질분해효소 활성측정을 수행하였다. 도체 pH와 온도는 사후 시간이 경과함에 따라 유의적으로 감소하였고(p<0.05), 사후 3시간에 6.50($31.99^{\circ}C$)을 나타내었다. $R_{248}$과 $R_{250}$ 값은 사후 9시간부터 유의적으로 증가하였지만, $R_{258}$ 값은 사후 9시간부터 감소하였다(p<0.05). Calpain I 활성은 사후 3시간부터 유의적으로 감소하였고, calpastatin 활성은 사후 9시간부터 유의적으로 감소하였다(p<0.05). pH와 도체온도는 $R_{258}$, calpain I, calpastatin에서 0.95이상의 높은 정의 상관관계를 나타내었지만, $R_{248}$과 $R_{250}$에서는 -0.97 이상의 부의 상관관계를 보였다. 이상의 결과를 종합하면, 사후9시간 이후에 대사 변화가 급격히 발생하는 것으로 나타났고, pH와 도체온도의 상관도가 높아 잠재적인 연화능력의 예측이 가능할 것으로 사료되지만, 사후 3시간부터 9시간 사이의 대사변화에 대하여 좀 더 심도 깊은 연구할 필요가 있을 것으로 생각된다.

• Asian-Australasian Journal of Animal Sciences
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• 제26권3호
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• pp.443-454
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• 2013

Tenderness is the most important meat quality trait, which is determined by intracellular environment and extracellular matrix. Particularly, specific protein degradation and protein modification can disrupt the architecture and integrity of muscle cells so that improves the meat tenderness. Endogenous proteolytic systems are responsible for modifying proteinases as well as the meat tenderization. Abundant evidence has testified that calpains (CAPNs) including calpain I (CAPN1) and calpastatin (CAST) have the closest relationship with tenderness in livestock. They are involved in a wide range of physiological processes including muscle growth and differentiation, pathological conditions and post-mortem meat aging. Whereas, Calpain3 (CAPN3) has been established as an important activating enzyme specifically expressed in livestock's skeletal muscle, but its role in domestic animals meat tenderization remains controversial. In this review, we summarize the role of CAPN1, calpain II (CAPN2) and CAST in post-mortem meat tenderization, and analyse the relationship between CAPN3 and tenderness in domestic animals. Besides, the possible mechanism affecting post-mortem meat aging and improving meat tenderization, and current possible causes responsible for divergence (whether CAPN3 contributes to animal meat tenderization or not) are inferred. Only the possible mechanism of CAPN3 in meat tenderization has been confirmed, while its exact role still needs to be studied further.

• 대한한의학회지
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• 제23권3호
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• pp.145-163
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• 2002

Objectives: The purpose of this investigation was to evaluate the effects of Woohwangcheongsim-won and to study the mechanism for neuronal death protection in hypoxia with Embryonic day 20 (E20) cortical cells of a rat (Sprague Dawley). Methods: E20 cortical cells were dissociated in neurobasal media and grown for 14 days in vitro (DIV). On 14 DIV, Woohwangcheongsim-won was added to the culture media for 24 hrs or 72 hrs. On 17 DIV, cells were given a hypoxic shock and further incubated in normoxia for another three days. On 20 DIV, Woohwangcheongsim-won's effects for neuronal death protection were evaluated by LDH assay, propidium iodide stain and phospho-H2AX immunostain and the mechanisms were studied by Bcl-2, Bak, Bax, caspase family, PKCα, ca1pain I. Results & Conclusions : 1. This study indicated that Woohwangcheongsim-won's effects for neuronal death protection in hypoxia were confirmed by LDH assay, propidium iodide stain and phospho-H2AX immunostain in culture method of Embryonic day 20(E20) cortical neuroblasts. 2. Woohwangcheongsim-won's mechanisms for neuronal death protection in hypoxia are to reduce the membrane damage fraction, to restrain DNA truncate, to restrain inflow of cytochrome c into cellularity caused by Bak diminution, to reduce the caspase cascade intiator caspase-8 and the effector caspase-3, to reduce the calpain I activity and to increase PKCand its activity in the membrane fraction. (J Korean Oriental Moo 2002;23(3):145～163)

• Asian-Australasian Journal of Animal Sciences
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• 제24권1호
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• pp.9-16
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• 2011

The calpain I (CAPN1) gene is an important marker for meat tenderness and marbling score in the bovine, but there were no studies to determine whether the CAPN1 gene had an association with other meat quality traits. In this study, we examined the relation between genetic polymorphisms of the CAPN1 gene and some meat quality traits in Yanbian Yellow Cattle of China. By PCRSSCP and gene sequencing in 321 unrelated Yanbian yellow cattle, twenty seven single nucleotide polymorphisms (SNPs) were detected in CAPN1, two existed SNPs in exon 8 and exon 17 resulted in the change of AA at F311S and M599V, respectively, and the otherpolymorphisms were at intron 7, 8, 14, 16 and 17. There were different preponderant genotypes at the corresponding gene locus and all genotypes were not associated with tenderness but other meat traits. This is the first study of the relationship between CAPN1 and meat quality besides tenderness in Yanbian yellow cattle of China.

• 대한약리학회지
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• 제29권2호
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• pp.165-174
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• 1993

칼슘이온은 신경섬유 성장의 중요한 조절인자이나 그 정확한 작용기전은 불명확하다. 세포골격 단백은 in vivo 및 in vitro에서 칼슘의존성 단백분해효소(칼파인)에 의해 신속히 분해되므로, 칼슘이온에 의한 신경섬유의 퇴행에 있어서 칼파인의 관련성을 추구하기위하여, 배양된 해마신경세포에서 칼슘이온 ionophore인 A23187에 의한 신경섬유의 성장억제가 칼파인의 억제제인 EST 및 MDL 28170에 의해 차단되는지를 조사하였다. A23187은 100nM의 농도에서 축삭에는 영향이 없이 수상돌기의 퇴행을 유발하였으나, 300 nM의 농도에서는 축삭의 성장을 억제하였다. EST(5 혹은 20 uM) 및 MDL 28170(20 uM)은 100 nM A23187의 수상돌기에 대한 작용과 300 nM A23187의 축삭에 대한 작용을 효과적으로 차단하였다. EST는 A23187에의한 세포내 칼슘이온의 증가를 차단하지 못하였다. 이상의 결과는 해마추상체세포에서 칼슘에 의한 신경섬유의 퇴행이 칼파인에 의해 매개됨을 시사한다.

• 한방재활의학과학회지
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• 제21권4호
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• pp.49-65
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• 2011

Objectives: This study was performed to investigate the effects of Gagamsosokmyeong-tang(Jiajianxiaoxuming-tang) on the monosodium iodoacetate(MIA) induced early state osteoarthritis in rats. Methods: Osteoarthritis was induced by injection of MIA(0.25 mg) into knee joints of rats. Osteoarthritis rats were divided into control(n=8) and treated=8 group respectively, Control group was taken distilled water and treated group was taken extracts of Gagamsosokmyeong-tang(Jiajianxiaoxuming-tang) by orally for 20 days. Body weight was measured at 0, 5, 10, 15, 20 days after MIA injection. At the end of experiment, gross and histopathological examination on the articular cartilages of the knee joints were performed. Proteoglycan(PG) content of articular cartilages were analysed by safranine O staining method. The content of tumor necrosis $factor-{\alpha}(TNF-{\alpha})$, $interleukin-1{\beta}(IL-1{\beta})$ in synovial fluids were analysed by enzyme-inked inmunosorbent assay(ELISA) method. And also cycloxygenase-2(COX-2), matrix metalloproteinase 3(MMP-3), inducible nitric oxide synthase(iNOS), calpain immunochistochemical examination on the knee joints were performed. Results: PG content in articular cartilages of the treated group was significantly increased compared with control group. Histopathological osteoarthritic score of the treated group was significantly decreased compared with control group. $TNF-{\alpha}$ content in synovial fluids, expression of iNOS and calpain in synovial membrane of the treated group were significantly decreased compared with control group. But body weight, $1L-1{\beta}$ content in synovial fluids, expression of iNOS and MMP-3 of the treated group were not significantly changed compared with control group. Conclusions: On the basis of these results, we conclude that Gagamsosokmyeong-tang(Jiajianxiaoxuming-tang) has anti-arthritic effects on the MIA induced early stage osteoarthritis in rats.

• Asian-Australasian Journal of Animal Sciences
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• 제19권8호
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• pp.1085-1089
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• 2006

The objectives of this study were to confirm a location of the calpastatin (CAST) gene in chromosome 2 and to detect associations of genetic variations with economic traits in the porcine CAST gene as a candidate gene for growth and meat quality traits in pigs. Calpastatin is a specific endogenous inhibitor of calpains. The calpain protease system is ubiquitous, and is involved in numerous growth and metabolic processes. Three single nucleotide variations were identified within a 1.6 kb fragment of the porcine CAST gene and these polymorphisms were used for genetic linkage mapping. Linkage and QTL mapping were performed with the National Livestock Research Institute (NLRI) reference families using eight microsatellites and SNP makers in the CAST gene. The porcine CAST gene was mapped adjacent to the markers, SW395 and SW1695 on SSC2 with LOD scores of 15.32 and 8.50, respectively. According to the QTL mapping, a significant association was detected at 82 cM between SW395 and CAST-Hinf I for weight at the age of 30 weeks. In addition, an association study was performed with the $F_2$ animals of NLRI reference families for Hinf I, Msp I and Rsa I polymorphisms in the CAST gene. Two polymorphisms, CAST-Rsa I and CAST-Hinf I, showed significant correlation for growth traits at p<0.01 and p<0.05, respectively.