• 한국식품영양과학회지
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• 제36권3호
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• pp.305-310
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• 2007

복분자, 산수유 및 토사자를 일정한 비율로 배합하여 열수추출로 얻어진 KH-305를 일반쥐에 투여해서 해면체 평활근 이완에 관련된 세포 내 신호전달체계 NO-cGMP pathway에 관여하는 NOS, 혈액내의 testosterone, BVSMCs cell에서 cGMP농도를 측정하여 음경발기 지속 및 촉진에 미치는 영향을 보았으며 음경조직의 활성산소제거와 관련하여 SOD/Mn, SOD/Cu의 단백질 발현정도를 측정하였다. KH-305는 NO-pathway에 관여하는 NOS의 발현증가, 낮은 농도에서의 cGMP농도 증가, testosterone의 수치를 증가시킴으로써 발기유지 및 촉진시키고, 동시에 음경조직내의 활성산소 및 NO 합성에서 나타나는 독성을 조절하여 주는 SOD발현이 증가됨으로써 활성산소에 의한 음경피로도를 경감시켜 음경해면체 평활근의 이완장애를 일으키는 발기부전 증상을 개선시킬 것으로 생각된다.

• Archives of Pharmacal Research
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• 제18권6호
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• pp.434-439
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• 1995

In pancreatic cells, NO formation is associated with increased levels of cGMP and endocrine/exocrine secretion. In the present study, the role of NO in the regulation of exocrine secretion was investigated in rat pancreatic tissues. Treatment of rat pancreatic tissue with sholecystokinin-pancreozymin (CCK-PZ) resulted in an significant increase in arginine conversion to citruline, the amount of nitrite/nitrate, the release of amylase, and the level of cGMP. Furthermore, CCK-PZ stimulated increase of amylase release and conversion of arginine to citrulline transformation were counteracted by the inhibitor of NO synthase, $N^G-nitro-L-arginine$ methyl ester. The results on the time course of CCK-PZ-induced citrulline formation within the first seconds of simulation. The kinetics of citrulline accumulation correlate well with those of cGMP rise, which further confirms the conclusion that NO mediates the response to CCK-PZ by cGMP.

• 한국축산식품학회지
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• 제20권2호
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• pp.159-165
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• 2000

Glycomacropeptide(GMP) was purified from cheese whey which is obtaining as a byproduct in cheese producing. Cheese whey was first concentrated 10 times with a ultrafiltration aparratus, and then heated at 95℃ for 5 min. The concentrated fraction was centrifuged at 20,000×g for 30 min to remove fat layer. The supernatant layer enriched GMP protein was fractionated by ion exchange chromatography on DEAE-Sepharose Fast Flow column. GMP was bound to DEAE resin and eluted with 0.1∼0.25 M NaCl when using a linear NaCl gradient from 0 M to 0.5 M. The purified GMP gave a single band of 24 kDa which seems to be trimer molecular weight in SDS-PAGE, and migrated to the same molecular weight with control GMP obtained commercially. Its amino acid composition were consistent with that of standard GMP. About 0.71 g of GMP was recovered from 1 L of cheese whey. These results indicate that glycomacropeptide could be simply purified from cheese whey by using ultrafiltration and DEAE column chromatography. 본 연구에서는 치즈 제조시의 부산물인 치즈 유청으로부터 glycomacropeptide(GMP)를 정제하려고 하였다. 치즈 유청을 한외여과 장치를 이용하여 10배로 농축하여 공시시료로 하였고, 95℃에서 5분간 가열한 후 20,000rpm에서 30분간 원심분리하였다. 원심분리 후 혼입된 지방층을 제거하였고, GMP를 포함하고 있는 상청액은 DEAE-Sepharose Fast Flow 크로마토그래피 칼럼에 주입하였다. GMP는 DEAE 수지의 관응기에 결합하였으며, 0에서 0.5M의 NaCl 직선농도 구배를 실시하였을 때 약 0.1∼0.25M에서 용출되었다. 정제한 GMP는 SDS-PAGE에서 단일한 band를 나타냈으며, 분자량은 24kDa으로 trimer 형태로 상업적으로 제공받은 대조 GMP와 같은 분자량 위치로 영동하였다. 정제한 GMP의 아미노산 조성은 대조GMP의 결과와 비교하였을 때 Ser, Val이 약간 적었으며 Gly은 반대로 2배의 함량을 나타냈으나 전체적인 조성비에서는 거의 일치하였다. 단백질의 회수율은 유청 1L에서 약 0.71g의 GMP를 효과적으로 분리한 것으로 나타났다. 이 결과로부터 한외여과와 DEAE-Sepharose Fast Flow chromatography를 이용하여 GMP를 산업적으로 대량 정제할 수 있는 것이 확인되었다.

• 대한수의학회지
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• 제43권4호
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• pp.615-624
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• 2003

Vascular smooth muscle relaxation is modulated by an increase in cGMP subsequent to nitric oxide (NO) production by endothelial cells. The effects of cAMP and cGMP phosphodiesterase (PDE) inhibitors were investigated in phenylephrine-precontracted rat aorta rings by using the specific inhibitors of PDE I, III, IV and V as relaxing agents (calmodulin-activated PDE inhibitors, IBMX and $W_7$, type I; cAMP-specific PDE inhibitors, milrinone, type IV; Ro 20-1724, type III and cGMP-specific PDE inhibitor, zaprinast, type V). All the PDE inhibitors produced a concentration-dependent relaxation in the ring with intact endothelium (+E). Except for milrinone, all the PDE inhibitors-induced relaxations were inhibited by removal of extracellular $Ca^{2+}$, $N^G$-nitro-L-arginine, $N^G$-nitro-L-arginine methyl ester, methylene blue (MS) or nifedipine. The specific PDE I and PDE IV inhibitors both produced endothelium-independent relaxations which were inhibited by MS in -E rings. However, zaprinast had no effect in -E rings. Except for milrinone, sodium nitroprusside (a NO donor)-induced relaxation was significantly augmented by all PDE inhibitors in +E rings. The results suggest that I) the vasorelaxant properties of IBMX, $W_7$, Ro 20-1724 and zaprinast are dependent on endothelium or on interaction with $Ca^{2+}$ regulation, 2) each PDE is differently distributed in vascular tissues (endothelial and smooth muscle cells), 3) the vasodilations of PDE inhibitors are due to the increase of cAMP and cGMP formation through inhibition of cAMP- and cGMP-PDE and 4) the vasodilation action of milrinone does not involve in endothelial-cyclic nucleotide system.

• 약학회지
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• 제59권1호
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• pp.40-46
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• 2015

The term "GMP" firstly came on the 1962 amendment of the Federal Food, Drug and Cosmetic (FD&C) Act and the US FDA established and officially announced the Good Manufacturing Practice Regulation for the first time in the world in 1963. In 1969, the World Health Organization published the GMP regulation and recommended that member states adopt the GMP regulation and implement the "GMP Certification Scheme" for international commerce of finished pharmaceutical products. As a result, GMP requirements have become important ones that have to be complied with in the manufacture of pharmaceutical products. The Korean GMP regulation was announced as the official notification by the Ministry of Health and Social Affairs in 1977. The KGMP regulation was voluntarily adopted by pharmaceutical companies at the early stage, but it had become mandatory. In addition, various kinds of GMP regulations have been established to cover active pharmaceutical ingredients, biological products and others, in addition to finished pharmaceutical products. Taking account of technological development and changes in the pharmaceutical environments, the KGMP regulation was fully amended and harmonized with GMP requirements of developed countries. In this way, the KGMP has developed to keep up with international trends and standards, leading to accession to the Pharmaceutical Inspection Cooperation Scheme (PIC/S).

• Journal of Microbiology and Biotechnology
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• 제18권7호
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• pp.1227-1234
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• 2008

Pseudomonas aeruginosa is an opportunistic human pathogen that produces and secretes exopolysaccharides (EPS), in which cells are embedded to form a highly organized community structure called biofilm. Here, we characterized the role of cyclic diguanylate (c-di-GMP) and EPS (PEL) overproduction in the wspF mutant phenotypes of P. aeruginosa PA14 (wrinkly appearance, hyperadherence, impaired motilities, and reduced virulence in acute infections). We confirmed that the elevated c-di-GMP level plays a key role in all the wspF mutant phenotypes listed above, as assessed by ectopic expression of a c-di-GMP-degrading phophodiesterase (PvrR) in the wspF mutant. In contrast, PEL EPS, which is overproduced in the wspF mutant, was necessary for wrinkly appearance and hyperadherence, but not for the impaired flagellar motilities and the attenuated virulence of the wspF mutant. These results suggest that c-di-GMP affects flagellar motility and virulence, independently of EPS production and surface adherence of this bacterium.

• Journal of Nutrition and Health
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• 제41권8호
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• pp.691-700
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• 2008

본 연구는 착상 후 단계의 쥐 배아와 난황낭을 대상으로 구리 결핍이 NO 하부 신호전달체계에 영향을 주는지를 알아보기 위한 것으로, 연구 결과는 다음과 같이 요약할 수 있다. 첫째, 구리 결핍은 정상적인 배아 및 난황낭 발달을 억제하고, NO의 생물학적 이용도와 아세틸콜린에 대한 NO dose-response를 낮추었다. 둘째, 구리 결핍은 NO의 하부 신호전달 물질인 cGMP 수준을 감소시켰으나, NO/cGMP 하부 신호전달체계 표적 중 하나인 P-VASP에는 영향을 미치지 않았다. 셋째, 구리 결핍 배양액에 NO donor를 첨가하는 것은 구리 결핍 배아와 난황낭의 기형 발생 빈도를 구리 정상군과 비슷한 수준으로 개선시켰다. 넷째, NO donor 첨가는 구리 결핍군에서 감소되었던 cGMP의 농도를 유의적으로 증가시켰지만, P-VASP에는 영향을 미치지 않았다. 상기 연구 결과들은 구리 결핍으로 인한 NO의 생물학적 이용도의 감소가 기형발생의 주요 발생 기전이라는 것을 뒷 받침하고 있다. 또한, 임상적으로 임신 기간 중 적절한 구리 섭취의 중요성을 강조한다.

• 생명과학회지
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• 제16권7호
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• pp.1225-1228
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• 2006

cAMP와 복합체를 형성한 cAMP 수용성 단백질은 DNA와 결합하여 ${\sim}90$도 정도의 예리한 DNA bending을 유도한다. 그러나 이전의 논문[5]에 의하면 돌연변이 CRP:cGMP 복합체는 돌연변이 CRP:cAMP 복합체보다 아크릴아미드 겔에서 상대적으로 빠른 이동속도를 보였다. CRP와 cyclic nucleotide 존재하에서 DNA의 구조 변화를 알아보기 위하여 6가지 준비된 DNA조각들을 사용하여 몰 고리화 인자(molar cyclization factor)[13]를 측정하였다. 이들 자료를 사용하여 nonlinear regression analysis를 통하여 cGMP 존재하에서 돌연변이 CRP는 DNA bending을 형성하지 않으나 CAMP 존재하에서 나선 꼬임과 같은 DNA 구조 변화없이 DNA bending을 형성한다.

• 대한약리학회지
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• 제31권3호
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• pp.291-297
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• 1995

본 연구의 목적은 protein kinase C의 활성물질인 phorbol ester의 수축에 대한 cGMP 및 cAMP의 조절기전을 명확히 하기 위하여 흰쥐의 대동맥을 재료로 실험을 수행하였다. Sodium nitroprusside는 guanylyl cyclase를 활성화시켜 cGMP를, forskolin은 adenylyl cyclase를 활성화시켜서 cAMP를 증가시키는 것으로 보고되어 있으므로 위의 두 약물을 선택하였다. Phorbol ester는 시간경과와 함께 지속적인 수축을 발생하였으며 30분경 안정상태에 도달하였다. 동시에 20-kDa myosin light chain (MLC)의 인산화도 증가하였으며 30분경 최대치를 나타내었다. Sodium nitroprusside와 forskolin은 phorbol ester에 의한 수축을 농도의존적으로 억제하였으나 sodium nitroprusside가 forskolin보다 더욱 민감하게 억제하였다. Phorbol ester는 $^{45}Ca^{2+}$의 유입을 증가시켰고 sodium nitroprusside와 forskolin은 이 증가된 $^{45}Ca^{2+}$을 유의하게 억제하였다. Phorbol ester에 의하여 증가된 MLC의 인산화는 sodium nitroprusside 및 forskolin 각각의 최대농도로 억제되었다. 이상과 같은 결과로 볼때 아마도 cGMP와 cAMP는 phorbol ester에 의한 수축을 $^{45}Ca^{2+}$ 유입억제에 이은 MLC 인산화 억제에 의하여 이완작용을 나타내는 것으로 추측되며 cGMP가 cAMP보다 protein kinase C 매개의 수축조절에 더 중요하게 작용하리라 추측된다.

• The Korean Journal of Physiology and Pharmacology
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• 제10권3호
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• pp.149-154
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• 2006

Selective inhibition of phosphodiesterase (PDE) 5 opened a new therapeutic approach for cardiovascular disorders. Therefore, the effect of PDE5 inhibition on the cardiac function should thoroughly be defined. The purpose of the present study was to define the effects of sildenafil, a selective inhibitor of PDE5, on the atrial cGMP efflux, atrial dynamics, and the release of atrial natriuretic peptide (ANP). By perfusing rabbit left atria to allow atrial pacing, changes in atrial stroke volume and pulse pressure, transmural extracellular fluid translocation, cGMP efflux, and ANP secretion were measured. SIN-I, an NO donor and soluble (s) guanylyl cyclase (GC) activator, and C-type natriuretic peptide (CNP), an activator of particulate (p) GC activator, were used. Sildenafil increased basal levels of cGMP efflux slightly but not significantly. Sildenafil in a therapeutic dose increased atrial dynamics (for atrial stroke volume, $2.84{\pm}1.71%$, n=12, vs $-0.71{\pm}0.86%$, n=21; p<0.05) and decreased ANP release ($-9.02{\pm}3.36%$, n=14, vs $1.35{\pm}3.25%$, n=23; p < 0.05), however, it had no effect on the SIN-1- or CNP-induced increase of cGMP levels. Furthermore, sildenafil in a therapeutic dose accentuated SIN-1-induced, but not CNP-induced, decrease of atrial pulse pressure and ANP release. These data indicate that PDE5 inhibition with sildenafil has a minor effect on cGMP levels, but has a distinct effect on pGC-cGMP- and sGC-cGMP-induced contractile and secretory function.