• The Korean Journal of Physiology
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• v.10 no.1
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• pp.1-5
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• 1976

The trypanocidal drug suramin, an impermeant polyanion, has been shown to be a powerful inhibitor of the calcium uptake and calcium-stimulated ATPase activity of sarcoplasmic reticulum (Fortes et al., 1974). In view of this finding, an attempt was made to investigate the effect of suramin on $Ca^{++}$ transport in resealed red cells and on $Ca^{++}$-activated ATPase in red blood cell membrane fragments (RBCMF). The results obtained are summarized as follows. 1. $Ca^{++}$ outflux from the resealed RBC was inhibited by suramin and the inhibitory action of suramin is proportional to the concentration of drug added inside the RBC preparation. When suramin is added both inside and outside the RBC preparation simultaneously, the magnitude of the inhibitory effect was more pronounced, suggesting that suramin inhibits both active $Ca^{++}-^{45}Ca$ exchange diffusion across the RBC membrane. 2. Suramin inhibits the $Ca^{++}$-activated ATPase of the RBCMF and the effect of inhibition by the drug was also concentration dependent. From the above results, it may be concluded that suramin inhibits $Ca^{++}$ transport across RBC membrane by inhibiting $Ca^{++}$-activated ATPase activity which has been known to be linked with active $Ca^{++}$ transport.

• Animal cells and systems
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• v.1 no.4
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• pp.641-646
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• 1997

Lysophosphatidic acid (LPA; 1-acyl-glycerol-3-phosphate) has been known as an intercellular phospholipid messenger with a wide range of biological activities. In this study, the effect of LPA on both the proliferation and differentiation of rat E63 myoblasts has been investigated. In the serum-free Insulin-Transferrin-Selenium (ITS) media, the proliferation of E63 cells was largely restricted. Addition of LPA into the ITS media strongly promoted the cell proliferation and resulted in two to four fold increase of cell number. Furthermore, it appeared to increase the percent fusion in a dose-dependent manner up to 15 ug/ml. The synthesis of myosin heavy chain (MHC) was increased by LPA as well. These results indicate that LPA is able to promote both cell proliferation and differentiation in rat E63 myoblasts. Suramin, known to have uncoupling activity on growth factor-receptor interaction, was tested for antagonistic activity in myoblast proliferation and differentiation. Myoblasts grown in the ITS medium containing LPA were able to proliferate well even in the presence high concentration of suramin whereas myoblast differentiation was completely blocked by 30 ug/ml of suramin. The inhibitory effect of suramin on the myoblast differentiation was completely reversible by removing the suramin. This result indicates that the intracellular signaling pathway of LPA leading to cell proliferation might be distinct from that leading to cell differentiation on E63 myoblasts. Also, the antagonistic effect of suramin suggests that the differentiation activity elicited by LPA might be mediated by a specific G protein-coupled receptor.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.12 no.4
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• pp.1770-1774
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• 2011

This paper was to characterize the transfer of diacylglycerol(DAG) from lipophorin to Manduca sexta larval fat bodies. $[^3H]$-DAG-labeled Lp($[^3H]$-DAG-Lp) was incubated with the larval fat bodies under different times and the time of DAG transfer was determined. Incubation of fat bodies with $[^3H]$-DAG-Lp resulted in accumulation of DAG and TAG in the tissue. The transfer of $[^3H]$-DAG was inhibited in the presence of suramin and unlabeled lipophorin, which would be consistent with a lipophorin receptor. The effects of suramin may be complex because it can change membrane properties when bound to the lipophorin receptor and affect the rate of DAG transfer. To investigate the lipid uptake via receptor-mediated endocytosis, we treated with endocytosis inhibitors, ammonium chloride and chloroquine. The results show that the transfer process of lipid by lipophorin and fat bodies is receptor-mediated endocytosis.

• Biomolecules & Therapeutics
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• v.14 no.1
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• pp.25-29
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• 2006

We tested effects of bioactive lysophospholipids including lysophosphatidic acid (LPA), lysophosphatidylcholine (LPC), sphingosylphosphorylcholine (SPC), and sphingosine I-phosphate (S1P) on membrane potential in C6 glioma cells to understand action mechanism of the lysophospholipids. Membrane potential was estimated by measuring fluorescence change of DiBAC-loaded glioma cells. LPA largely increased membrane potential and the increase was gradually diminished. LPC also increased the membrane potential, however, the increase sustained. SPC induced smaller increase of membrane potential than LPC. SIP was not able to change the membrane potential. We tested effects of suramin and pertussis toxin on lysophospholipid-induced membrane potential increase. However, there wasn't any effect. The membrane potential increase was partially diminished in $Na^+$-free media, suggesting $Na^+$ influx as a component of membrane potential changes. Thus, involvement of $Na^+$ influx in the increase of membrane potential by lysophospholipids and independence of suramin-sensitive GPCRs and pertussis toxin-sensitive G proteins are found in this study.

• The Korean Journal of Physiology and Pharmacology
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• v.2 no.4
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• pp.419-425
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• 1998

To clarify the effect of extracellular ATP in cultured C6 glioma cells, ATP-induced cytosolic free calcium ($[Ca^{2+}]_i$) mobilization and cell proliferation were investigated. ATP-induced $[Ca^{2+}]_i$ increased in a dose-dependent manner $(10^{-7}\;M{\sim}10^{-3}\;M)$. ATP-induced $[Ca^{2+}]_i$ increases were slightly slowed in extracellular calcium-free conditions especially in sustained phase. ATP-induced $[Ca^{2+}]_i$ increment was also inhibited by the pretreatment of U73122, a phospholipase C (PLC) inhibitor, in a time-dependent manner. Suramin, a putative $P_{2Y}$ receptor antagonist, dose-dependently weakened ATP-induced $[Ca^{2+}]_i$ mobilization. Significant increases in cell proliferation were observed at 2, 3, and 4 days after ATP was added. Stimulated cell proliferation was also observed with adenosine at days 2 and 3. This cell proliferation was significantly inhibited by the treatment with suramin. Ionomycin also stimulated cell proliferation in a concentration-dependent manner. In conclusion, we suggest that extracellular ATP stimulates C6 glioma cell proliferation via intracellular free calcium mobilization mediated by purinoceptor.

• Proceedings of the KAIS Fall Conference
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• 2006.11a
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• pp.287-289
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• 2006

곤충 혈림프에서 존재하는 리포포린은 선택적으로 지질을 지질 사용및 저장기관으로 운반한다. 본 연구는 유충지방체, 성충난소 및 정소로 지질의 운반과 유충지방체 및 성충난소로 리포포린 자체가 흡수되는 과정을 조사하였다. 이들의 기능을 조사하기 위해 FITC-labeled 리포포린과 DiI-labeled 리포포린을 사용하였다. 유충지방체, 성충난소 및 정소를 DiI-labeled 리포포린과 배양한 결과 리포포린으로 부터 각 기관으로 지질을 운반함을 알 수 있었고, 또한 receptor-mediated endocytosis 억제제인 suramin, unlabeled 리포포린과 배양한 결과는 리포포린에서 각 기관으로 운반되는 지질의 양이 현저하게 감소함을 알 수 있었다. 또한, 유충지방체와 성충난소를 FITC-labeled 리포포린과 배양한 결과 위에서 언급한 지질 뿐만 아니라 리포포린 자체도 각 기관의 에너지원으로 사용하기 위해 흡수된다는 사실을 알 수 있었으며, suramin과 unlabeled 리포포린과 배양한 결과 리포포린 자체가 흡수되는 양이 현저하게 감소함을 알 수 있었다. 위 실험결과로부터 리포포린에 의한 지질의 운반과정과 리포포린 자체의 흡수과정이 receptor-mediated endocytosis로 이루어짐을 알 수 있었다.

• The Korean Journal of Pesticide Science
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• v.5 no.3
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• pp.41-46
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• 2001

This study was conducted to develop a high throughput system for screening acetolactate synthase(ALS) inhibitors, and to detect basic mother molecules for developing new novel herbicide candidates. The high throughput screening (HTS) method using 96-well plate and microplate reader was developed. This method is 8 times more effective than basic technique in one cycle per person. Futhermore, considering for less than 1/10 volume of materials required for ALS test and enzyme kinetics with 16 times faster speed compared to those of former procedure, this HTS method has more than 100 times higher efficacy than basic system in a consecutive procedure. We discovered 11 new ALS inhibitors such as 2-oxoglutaric acid, aminooxyacetic acid, azelaic acid, citric acid, cyanuric fluoride, itaconic acid, malonic acid, niclosamide, oxalic acid, glyoxylic acid, and suramin from 107 commercial plant-specific inhibitors using this technique. We hope these results might be useful to discover lead compounds for developing new novel herbicide candidate.

• Journal of Acupuncture Research
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• v.20 no.1
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• pp.177-190
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• 2003

Objective : This study was intended to investigate the analgesic effects of electroacupuncture(EA) on mechanical allodynia according to the frequency and intensity of EA. Also to know if mechanical allodynia and the analgesic effects of EA is related to the sympathetci nervous system and/or the purinergic system. Methods : mechanical allodynia-induced rats were produced by resecting S1-S2 nerve. The zusanli(ST36) was used for acupoint and the rats were divided into 4 groups. Each group was given different stimuli[low frequency low intensity-EA(LFLI-EA), low frequency high intensity-EA(LFHI-EA), high frequency low intensity-EA(LFHI-EA), high frequency high intensity-EA(HFHI-EA)]. Futhermore, to make sympathectomy6-OHDA and phentolamine were administered intraperitonially and the concentration of norepinephrine(NE) were measured. As a ATP blocker, suramin was applied for this study. Results : Comparing to control group, each of the 4 groups(LFLI-EA, LFHI-EA, HFLI-EA, HFHI-EA) showed a significant reduction of response frequency of mechanical allodynia. LFHI-EA was more effective than that of LFLI-EA. The LFHI-EA group also had longer lasting effects from the stimulation than the other groups. Sympathectomy didn't show any reduction of response frequency of mechanical allodynia.(Each n=6, n=4). Nor did both sympathectomy and ATP block. The response frequency wasn't reduced by sympathectomy or by sympathectomy and ATP block, but was significantly reduced with LFHI-EA Conclusions : These results suggest that EA has a significant analgesic effect on mechanical allodynia which has no connection with NE and/or ATP.

• Archives of Pharmacal Research
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• v.24 no.6
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• pp.552-556
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• 2001

Histamine and arachidonic acid (AA) release was measured using the P2-purinoceptor antaongists, phospholipase $A_2{\;}(PLA_2)$ and cyclooxygenase (COX)/lipoxygenase (LOX) inhibitors to determine whether or not ATP-induced histamine release is associated with arachidonic acid (AA) release in rat peritoneal mast cells. ATP increased histamine release in a dose dependent manner, whereas adenosine did not. PPADS (a selective P2X-purinoceptor antagonist) and suramin (a nonselective P2X,2Y-purinoceptor antagonist) inhibited ATP-induced histamine release in a dose dependent manner. However, RB-2 (a P2Y-purinoceptor antagonist) did not block ATP-induced histamine release. Manoalide and oleyloxyethyl phosphorylcholine (OPC), secretory PLA$_2$ inhibitors, also inhibited ATP-induced histamine release dose-dependently. Both COX inhibitors (ibuprofen and indomethacin) and LOX inhibitors (baicalein and caffeic acid) inhibited ATP-induced histamine in a dose dependent manner. ATP significantly increased [$^3H$]AA release by 54%. PPADS and suramin significantly inhibited ATP-induced [3H]Ph release by 81% and 39%, respectively. ATP-induced histamine release was significantly inhibited by a variety of protein kinase inhibitors, such as bisindolmaleimide, genistein, methyl 2,5-dihydroxycinnamate, W-7 and trifluoperazine. Overall, the results suggest that ATP-induced histamine release is in part related to the PLA2-mediated AA metabolism and P2X-purinoceptors.

• International Journal of Oral Biology
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• v.31 no.4
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• pp.141-148
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• 2006

The effects of adenosine triphosphate(ATP) on salivary glands have been recognized since 1982. The presence of purinergic recepetors(P2Rs) that mediate the effects of ATP in various tissues, including parotid and submandibular salivary gland, has been supported by the cloning of receptor cDNAs and the expression of the receptor proteins. P2Rs have many subtypes, and the activation of these receptor subtypes increase intracellular $Ca^{2+}$, a key ion in the regulation of the secretion in the salivary gland. The apical pores of taste buds in circumvallate and foliate papillae are surrounded by the saliva from von Ebner salivary gland(vEG). Thus, it is important how the secretion of vEG is controlled. This study was designed to elucidate the roles of P2Rs on salivary secretion of vEG. Male Sprague-Dawley rats (about 200 g) were used for this experiment. vEG-rich tissues were obtained from dissecting $500-1,000\;{\mu}m$ thick posterior tongue slices under stereomicroscope view. P2Rs mRNA in vEG acinar cells were identified with RT-PCR. To observe the change in intracellular $Ca^{2+}$ activity, we employed $Ca^{2+}-ion$ specific fluorescence analysis with fura-2. Single acinar cells and cell clusters were isolated by a sequential trypsin/collagenase treatment and were loaded with $10\;{\mu}M$ fura -2 AM for 60 minutes at room temperature. Several agonists and antagonists were used to test a receptor specificity. RT-PCR revealed that the mRNAs of $P2X_4$, $P2Y_1$, $P2Y_2$ and $P2Y_3$ are expressed in vEG acinar cells. The intracellular calcium activity was increased in response to $10\;{\mu}M$ ATP, a P2Rs agonist, and 2-MeSATP, a $P2Y_1$ and $P2Y_2R$ agonist. However, $300\;{\mu}M\;{\alpha}{\beta}-MeATP$, a $P2X_1$ and $P2X_3R$ agonist, did not elicit the response. The responses elicited by $10\;{\mu}M$ ATP and UTP, a $P2Y_2R$ agonists, were maintained when extracellular calcium was removed. $10\;{\mu}M$ suramin, a P2XR antagonist, and reactive blue 2, a P2YR antagonist, partially blocked ATP-induced response. However, when extracellular calciums were removed, suramin did not abolish the responses elicited by ATP. These results suggest that P2Rs play an important role in salivary secretion of vEG acinar cells and the effects of ATP on vEG salivary secretion may be mediated by $P2X_4$, $P2Y_1$, $P2Y_2$, and/or $P2Y_3$.