• Applied Biological Chemistry
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• 제48권3호
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• pp.212-216
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• 2005

Thapsigargin은 동물조직에서 ER/SR-type $Ca^{2+}-ATPase$의 선택적 저해제로서, 토마토 뿌리조직으로부터 분리한 마이크로솜에서 ATPase의 특성을 조사하기 위하여 사용되었다. Thapsigargin은 마이크로솜 ATPase 활성을 농도의존적으로 저해하였으며, $10\;{\mu}M$ 농도에서 총활성의 약 30%를 저해하였다. 이것은 뿌리조직에서 $Ca^{2+}-ATPase$의 활성이 매우 낮다는 것을 고려할 때, thapsigargin이 뿌리조직의 주된 ATPase 활성인 원형질막 및 액포막의 $H^+-ATPase$ 활성을 저해할 가능성을 보인다. Thapsigargin의 효과는 ${NO_3}^-$를 사용하여 액포막 $H^+-ATPase$ 활성을 저해하였을 때 현저하게 감소하였다. 그러나, thapsigargin의 효과는 원형질막의 $H^+-ATPase$ 활성에는 영향을 미치지 않아, thapsigargin이 토마토 뿌리조직에서 액포막 $H^+-ATPase$를 선택적으로 저해함을 보여준다.

• 대한의생명과학회지
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• 제15권4호
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• pp.273-279
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• 2009

Cordycepin (3'-deoxyadenosine) is an adenosine analogue isolated from Cordyceps militaris, and it has been used as an anti-cancer and anti-inflammation ingredient in traditional Chinese medicine. We investigated the effects of cordycepin on human platelet aggregation induced by thapsigargin, and determined the cytosolic free $Ca^{2+}$ levels ($[Ca^{2+}]_i$), an aggregation-stimulating factor. Cordycepin significantly inhibited thapsigargin-induced platelet aggregation. Its inhibitory effect was continually sustained at the maximal aggregation concentration of thapsigargin. The thapsigargin-induced $[Ca^{2+}]_i$ were clearly reduced by cordycepin in the presence of exogenous $CaCl_2$ or extracellular $Ca^{2+}$-chelator (EDTA). These results suggest that cordycepin inhibited thapsigargin-induced $Ca^{2+}$-influx from extracellular domain and thapsigargin-induced $Ca^{2+}$-mobilization from intracellular $Ca^{2+}$ storage. Accordingly, our data demonstrated that cordycepin may have a beneficial effect on platelet aggregation-mediated thrombotic diseases by inhibiting a $[Ca^{2+}]_i$-elevation.

• 동의생리병리학회지
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• 제23권6호
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• pp.1368-1378
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• 2009

Unfolded protein response (UPR) is an important genomic response to endoplasmic reticulum (ER) stress. The ER response is characterized by changes in specific proteins, induction of ER chaperones and degradation of misfolded proteins. Also, the pathogenesis of several diseases like Alzheimer's disease, neuronal degenerative diseases, and diabetes reveal the role of ER stress as one of the causative mechanisms. Borneolum has been used for neuronal disease in oriental medicine. In the present study, the protective effect of borneolum on thapsigargin-induced apoptosis in rat C6 glial cells. Treatment with C6 glial cells with 5 uM thapsigargin caused the loss of cell viability, and morphological change, which was associated with the elevation of intracellular $Ca^{++}$ level, the increase in Grp78 and CHOP and cleavage of pro-caspase 12 Furthermore, thapsigargin induced Grp98, XBP1, and ATF4 protein expression in C6 glial cells. Borneolum reduced thapsigargin-induced apoptosis through ER pathways. In the ER pathway, borneolum attenuated thapsigargin-induced elevations in Grp78, CHOP, ATF4, and XBP1 as well as reductions in pro-caspase 12 levels. Also, our data showed that borneolum protected thapsigargin-induced cytotoxicity in astrocytes from rat (P3) brain. Taken together, our data suggest that borneolum is neuroprotective against thapsigargin-induced ER stress in C6 glial cells and astrocytes. Accordingly, borneolum may be therapeutically useful for the treatment of thapsigargin-induced apoptosis in central nervous system.

• Bulletin of the Korean Chemical Society
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• 제32권9호
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• pp.3469-3472
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• 2011

• BMB Reports
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• 제33권3호
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• pp.202-207
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• 2000

The depletion of intracellular calcium stores by thapsigargin treatment evoked extracellular calcium-dependent membrane currents in Xenopus laevis oocytes. These currents have been compared to those evoked by microinjection of a calcium influx factor (CIF) purified from Jurkat T lymphocytes. The membrane currents elicited by thapsigargin treatment (peak current, $163{\pm}60$ nA) or CIF injection (peak current, $897{\pm}188$ nA) were both dependent on calcium entry, based on their eradication by the removal of extracellular calcium. The currents were, in both cases, attributed primarily to well-characterized $Ca^{2+}-dependent$ $Cl^-$ currents, based on their similar reversal potentials (-24 mV vs. -28 mV) and their inhibition by niflumic acid (a $Cl^-$ channel blocker). Currents induced by either thapsigargin treatment or CIF injection exhibited an identical pattern of inhibitory sensitivity to a panel of lanthanides, suggesting that thapsigargin treatment or CIF injection evoked $Cl^-$ currents by stimulating calcium influx through pharmacologically identical calcium channels. These results indicate that CIF acts on the same calcium entry pathway activated by the depletion of calcium stores and most lanthanides are novel pharmacological tools for the study of calcium entry in Xenopus oocytes.

• Journal of Microbiology and Biotechnology
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• 제17권7호
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• pp.1134-1138
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• 2007

Cordycepin (3'-deoxyadenosine) is an adenosine analog, isolated from Cordyceps militaris, and it has been used as an anticancer and anti-inflammation ingredient in traditional Chinese medicine. We investigated the effects of cordycepin (3'-deoxyadenosine) on human platelet aggregation, which was induced by thapsigargin, a tumor promoter, and determined the cytosolic free $Ca^{2+}$ levels ($[Ca^{2+}]_i$) (an aggregation-stimulating molecule) and cyclic-guanosine monophosphate (cGMP) (an aggregation-inhibiting molecule). Cordycepin inhibited thapsigargin-induced platelet aggregation in a dose-dependent manner, and it clearly reduced the levels of $[Ca^{2+}]_i$, which was increased by thapsigargin ($1\;{\mu}M$) or U46619 ($3\;{\mu}M$). Cordycepin also increased the thapsigargin-reduced cGMP levels. Accordingly, our data demonstrated that cordycepin may have a beneficial effect on platelet aggregation-mediated thrombotic diseases through the $[Ca^{2+}]_i$-regulating system such as cGMP.

• The Korean Journal of Physiology and Pharmacology
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• 제20권4호
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• pp.387-397
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• 2016

Neurofibrillary tangles (NFTs) of microtubule-associated protein tau are a pathological hallmark of Alzheimer's disease (AD). Endoplasmic reticulum (ER) stress has been known to be involved in the pathogenesis of AD. However, the exact role of ER stress in tau pathology has not yet been clearly elucidated. In present study, the possible relationship between tau pathology and ER stress was examined in terms of sorcin, which is a calcium binding protein and plays an important role in calcium homeostasis. Our previous yeast two hybrid study showed that sorcin is a novel tau interacting protein. Caspase-3-cleaved tau (T4C3) showed significantly increased tau-sorcin interaction compared to wild type tau (T4). Thapsigargin-induced ER stress and co-expression of constitutively active $GSK3{\beta}$ ($GSK3{\beta}-S9A$) also exhibited significantly increased tau-sorcin interactions. T4C3-expressing cells showed potentiated thapsigargin -induced apoptosis and disruption of intracellular calcium homeostasis compared to T4-expressing cells. Overexpression of sorcin significantly attenuated thapsigargin-induced apoptosis and disruption of calcium homeostasis. In contrary, siRNA-mediated knock-down of sorcin showed significantly increased thapsigargin-induced apoptosis and disruption of calcium homeostasis. These data strongly suggest that sequestration of sorcin by aberrant forms of tau compromises the function of sorcin, such as calcium homeostasis and cellular resistance by ER stress, which may consequently result in the contribution to the progression of AD.

• IMMUNE NETWORK
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• 제12권6호
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• pp.253-260
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• 2012

${\alpha}$-Mangostin is a xanthon derivative contained in the fruit hull of mangosteen (Garcinia mangostana L.), and the administration of ${\alpha}$-Mangostin inhibited the growth of transplanted colon cancer, Her/CT26 cells which expressed Her-2/neu as tumor antigen. Although ${\alpha}$-Mangostin was reported to have inhibitory activity against sarco/endoplasmic reticulum $Ca^{2+}$ ATPase like thapsigargin, it showed different activity for autophagy regulation. In the current study, we found that ${\alpha}$-Mangostin induced autophagy activation in mouse intestinal epithelial cells, as GFP-LC3 transgenic mice were orally administered with 20 mg/kg of ${\alpha}$-Mangostin daily for three days. However, the activation of autophagy by ${\alpha}$-Mangostin did not significantly increase OVA-specific T cell proliferation. As we assessed ER stress by using XBP-1 reporter system and phosphorylation of $eIF2{\alpha}$, thapsigargin-induced ER stress was significantly reduced by ${\alpha}$-Mangostin. However, coadministration of thapsigargin with ${\alpha}$-Mangostin completely blocked the antitumor activity of ${\alpha}$-Mangostin, suggesting ER stress with autophagy blockade accelerated tumor growth in mouse colon cancer model. Thus the antitumor activity of ${\alpha}$-Mangostin can be ascribable to the autophagy activation rather than ER stress induction.

• Molecules and Cells
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• 제21권3호
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• pp.356-359
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• 2006

The transcription factor E2F1 coordinates cell cycle progression and induces apoptosis in response to DNA damage stress. Aside from DNA damage, the role of E2F1 in the endoplasmic reticulum (ER) stress signaling pathways is unclear. We found that $E2F1^{-/-}$ murine embryonic fibroblasts (MEFs) are resistant to apoptosis triggered by the ER stress inducer thapsigargin. In addition, E2F1 deficiency results in enhanced phosphorylation of eukaryotic translation initiation factor $2{\alpha}$ ($elF2{\alpha}$). These results therefore indicate that E2F1 deficiency increases phosphorylation of $elF2{\alpha}$ in response to ER stress triggered by thapsigargin, and suggest that the reduction in ER stress-induced apoptosis in E2F1-deficient cells is related to the high level of $elF2{\alpha}$ phosphorylation.

• 대한의생명과학회지
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• 제27권3호
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• pp.170-176
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• 2021

Thapsigargin (TG), a sarco/endoplasmic reticulum (ER) Ca²⁺-ATPase (SERCA) inhibitor, has been widely used as an agonist for platelet aggregation for decades. In this study, we investigated the effect of TG on the release of lactate dehydrogenase (LDH) for platelets and elucidated its mechanism. Platelet LDH release and platelet aggregation were increased by TG treatment; 1,000 nM of TG induced the complete lysis of platelets. Other agonists such as collagen (2.5 ㎍/mL), thrombin (0.1 U/mL), and ADP (10 mM) did not induce significant platelet LDH release despite platelet aggregation. Finally, we investigated the effects of pharmacological inhibitors on TG-induced platelet aggregation and LDH release. SP600125, a JNK inhibitor, and LY294002, a PI-3K inhibitor, inhibited TG-induced platelet LDH release but not platelet aggregation. Forskolin, an adenylyl cyclase activator, also inhibited LDH release without affecting platelet aggregation by TG. These results suggest that the TG-induced platelet aggregation was accompanied by LDH release but regulated by a different signaling pathway.