• Journal of Ginseng Research
• /
• 제37권3호
• /
• pp.361-370
• /
• 2013

A lysine histidine transporter (LHT) cDNA was isolated and characterized from the roots of Panax ginseng, designated PgLHT. The cDNA is 1,865 bp with an open reading frame that codes for a protein with 449 amino acids and a calculated molecular mass of 50.6 kDa with a predicted isoelectric point of 8.87. Hydropathy analysis shows that PgLHT is an integral membrane protein with 9 putative membrane-spanning domains. Multiple sequence alignments show that PgLHT shares a high homology with other plant LHTs. The expression profile of the gene was investigated by real-time quantitative polymerase chain reaction during various chemical treatments. PgLHT was up-regulated in the presence of abscisic acid, salicylic acid, methyl jasmonate, NaCl, and amino acids. To further explore the function of PgLHT gene, full-length cDNA of PgLHT was introduced into P. ginseng by Agrobacterium rhizogenes A4. The overexpression of PgLHT in the hairy roots led to an obviously increase of biomass compared to the controls, and after addition of the amino acids, the overexpressed-PgLHT hairy roots grew more rapidly than untreated controls during early stage of the culture cycle. The results suggested that the PgLHT isolated from ginseng might have role in the environmental stresses and growth response.

• Natural Product Sciences
• /
• 제18권4호
• /
• pp.211-214
• /
• 2012

Since 15-hydroxyprostaglandin dehydrogenase (15-PGDH) is the key metabolic enzyme of prostaglandin $E_2$ ($PGE_2$), inhibition of 15-PGDH is supposed to facilitate various physiological functions by increasing $PGE_2$. Methanol extract of Artemisia argyi (AAME) inhibited 15-PGDH ($IC_{50}$: $13.13{\mu}g/mL$) with relatively low cytotoxicity ($IC_{50}$: $415.00{\mu}g/mL$) and elevated extracellular $PGE_2$ levels in HaCaT cells. Real-time PCR analysis showed that AAME decreased significantly mRNA expression of PG transporter (PGT) in HaCaT cells. These results indicate that AAME could be applicable to functional materials as a 15-PGDH inhibitor and PGT expression inhibitor for the upregulation of extracellular $PGE_2$ level.

• Journal of Plant Biotechnology
• /
• 제49권3호
• /
• pp.171-177
• /
• 2022

The precise homeostatic regulation of local auxin accumulation in xylem precursors of cambium stem cell tissues is one of the most important mechanisms for plant vascular patterning and radial secondary growth. Walls are thin (WAT1), a novel intracellular auxin transporter, contributes directly to the auxin accumulation maxima in xylem precursors. According to recent research, the auxin signaling activated pathway-related gene network was significantly enriched during the secondary growth of Panax ginseng storage roots. These imply that during P. ginseng root secondary growth, specific signaling mechanisms for local auxin maxima in the vascular cambial cells are probably triggered. This study identified four WAT1-like genes, PgWAT1-1/-2 and PgWAT2-1/-2, in the P. ginseng genome. Their expression levels were greatly increased in nitratetreated storage roots stimulated for secondary root growth. PgWAT1-1 and PgWAT2-1 were similar to WAT1 from Arabidopsis and tomato plants in terms of their subcellular localization at a tonoplast and predicted transmembrane topology. We discovered that overexpression of PgWAT1-1 and PgWAT2-1 was sufficient to compensate for the secondary growth defects observed in slwat1-copi loss of function tomato mutants. This critical information from the PgWAT1-1 and PgWAT2-1 genes can potentially be used in future P. ginseng genetic engineering and breeding for increased crop yield.

• Asian-Australasian Journal of Animal Sciences
• /
• 제30권5호
• /
• pp.643-652
• /
• 2017

Objective: Prostaglandins (PGs) function in various reproductive processes, including luteolysis, maternal pregnancy recognition, conceptus development, and parturition. Our earlier study has shown that PG transporters ATP-binding cassette, subfamily C, member 4 (ABCC4) and solute carrier organic anion transporter family, member 2A1 (SLCO2A1) are expressed in the uterine endometrium in pigs. Since several other PG transporters such as ABCC1, ABCC9, SLCO4C1, and SLCO5A1 are known to be present in the uterine endometrium, this study investigated the expression of these PG transporters in the porcine uterine endometrium and placenta. Methods: Uterine endometrial tissues were obtained from gilts on day (D) 12 and D15 of the estrous cycle and days 12, 15, 30, 60, 90, and 114 of pregnancy. Results: ABCC1, ABCC9, SLCO4C1, and SLCO5A1 mRNAs were expressed in the uterine endometrium, and levels of expression changed during the estrous cycle and pregnancy. Expression of ABCC1 and ABCC9 mRNAs was localized mainly to luminal and glandular epithelial cells in the uterine endometrium, and chorionic epithelial cells during pregnancy. Conceptuses during early pregnancy and chorioallantoic tissues from mid to late pregnancy also expressed these PG transporters. $Estradiol-17{\beta}$ increased the expression of ABCC1 and SLCO5A1, but not ABCC9 and SLCO4C1 mRNAs and increasing doses of $interleukin-1{\beta}$ induced the expression of ABCC9, SLCO4C1, and SLCO5A1 mRNAs in endometrial explant tissues. Conclusion: These data showed that several PG transporters such as ABCC1, ABCC9, SLCO4C1, and SLCO5A1 were expressed at the maternal-conceptus interface, suggesting that these PG transporters may play an important role in the establishment and maintenance of pregnancy by regulating PG transport in the uterine endometrium and placenta in pigs.

• 약학회지
• /
• 제48권4호
• /
• pp.247-253
• /
• 2004

We investigated the hypoglycemic effect of formula containing Euonymus alatus (EA) and Mori Folium (MF) in multiple low dose (MLD) streptozotocin (STZ)-induced diabetic rats. In order to iduce hyperglycemic state 25 mg/kg of STZ was injected intraperitoneally for 5 consecutive days. SD rats were randomly divided into diabetic control and treatment groups. Treatment groups were administered with either 250 mg/kg of EA and 250 mg/kg of MF (E1Ml), or 500 mg/kg of EA mixed with same dose of MF (E2M2) for 3 weeks. Blood glucose levels and body weights were measured every 5th or 6th day. E1Ml and E2M2 both significantly reduced food intake, water intake, and fasting blood and urine glucose levels as compared to those in diabetic control group in a dose dependent manner. Body weight in diabetic control group was increased slightly after 3 weeks. Treatment group, however, showed gradual increase in body weights during 3 week-period. While plasma insulin levels of the diabetic control group were decreased to the level of 387$\pm$14 pg/ml from 534$\pm$36 pg/ml, those levels in E1Ml and E2M2-treated groups were both markedly increased by 13% and 26%, respectively. Urine glucose levels in E1Ml and E2M2-treated groups were also remarkably reduced by 17 and 26% compared to the levels of diabetic control group. While expression of membrane-bound glucose transporter-4 (GLUT-4) protein in skeletal muscle was reduced by 45% in diabetic control compared to the normal control, GLUT-4 protein expressions in E1Ml and E2M2-treated groups were augmented by 2 and 3.5 times compared to the diabetic control, respectively. Pancreatic HE staining experiments showed that E2M2-treated group revealed much less infiltrated mononuclear cells, indicating that E2M2 efficiently blocked insulitis induced by multiple low dose streptozotocin. Taken together, we conclude that formula containing EA and MF may prevent or delay the development of hyperglycemia through overexpression of GLUT-4 protein in skeletal muscle and prevention of insulitis.

• 한국약용작물학회지
• /
• 제22권6호
• /
• pp.457-462
• /
• 2014

Prostaglandin (PG) $E_2$ is an important mediator of skin wound healing without excessive scarring and gastric ulcer healing. However, $PGE_2$ has a short lifetime in vivo because it is metabolized rapidly by 15-hydroxyprostaglandin dehydrogenase (15-PGDH). Ethanol extract of Eriobotryae folium (EFEE) elevated intracellular and extracellular $PGE_2$ levels in HaCaT cells and inhibited 15-PGDH ($ED_{50}$ : $168.4{\mu}g/mL$) with relatively low cytotoxicity ($IC_{50}$ : $250.0{\mu}g/mL$). Real-time PCR analysis showed that mRNA expression of cyclooxygenase (COX)-1 and COX-2 enzymes were increased and prostaglandin transporter (PGT) was decreased in HaCaT cells by EFEE. Moreover, wound healing effect of EFEE ($168.4{\mu}g/mL$) was comparable to that of TGF-${\beta}1$ (300 pg/mL) as a positive control. These results demonstrate that EFEE may be valuable therapeutic materials for the treatment of $PGE_2$ level dependent diseases.

• 대한핵의학회지
• /
• 제38권2호
• /
• pp.180-189
• /
• 2004

다약제내성이 발현된 암세포에서 세포내의 항암제를 세포외로 배출시키는 기전을 체내에서 비침습적인 방법으로 영상화 할 수 있는 SPECT와 PET는 악성종양의 진단과 평가에 중요한 역할을 할 것으로 판단되나, 아직까지도 Pgp와 MRP의 운반능을 적절하게 평가하는 핵의학적 영상방법을 정립하는데는 극복해야할 문제점들이 많다. 지금까지의 MDR영상에 관한 연구들은 대부분이 $^{99m}Tc$-표지 방사성의약품을 이용한 연구였으나, PET의 임상 응용이 증가함에 따라 보다 특이적이고 쉽게 응용될 수 있는 PET용 방사성 추적자의 개발도 이루어져야 할 것이다. $^{99m}Tc$-MIBI의 암 세포내 일방향(unidirectional) 섭취는 음성인 세포막 전하와 세포내 소립체 기질 전하에 의하여 결정되므로, MIBI의 섭취는 다른 지용성 양전하를 띤 막전위 추적자들과 유사하게 작용한다. $^{99m}Tc$-표지 방사성의약품은 암조직의 혈류 증가나 소립체 용적이나 활성도가 증가하면 섭취가 증가할 수 있어 보다 특이적인 MDR추적제의 개발이 필요한 것이다. 최근 Lorke 등은 약제감수성 및 내성 인체대장암세포인 $HT-29^{par}$ 세포와 $HT-29^{mdrl}$ 세포를 이용한 연구에서, 두세포 모두에서 $^{18}F$-FDG의 섭취가 있었고, MDR이 발현된 세포와 종양에서 $^{18}F$-FDG 섭취가 훨씬 낮았고, MIBI는 MDR이 없는 모세포에서도 매우 낮았음을 보고한 바 있다. 이 세포는 전자현미경검사에서 사립체가 풍부하지 않은 세포였다. 그러므로 이러한 결과로 보아 $^{99m}Tc$-MIBI 영상에서 종양이 보이지 않거나 섭취가 미약하다고 해도 MDR이 발현되었다고 단정할 수는 없게 된다. 즉 MDR의 발현유무를 정확하게 감별할 수 있기 위하여는 저항이 없는 세포에 MIBI가 충분하게 섭취되어야 한다는 것이 필수적인 요건이며, 종양세포 종류에 따라서는 FDG가 MDR의 marker가 될 수 있다는 것이다. Pgp 수송체는 ATP의존성 약제배출 펌프이므로 MDR세포는 에너지가 많이 필요하여, MDR 세포는 당분해율(rate of glycolysis)이 증가되어 있고 HT-29 mdrl 종양세포에서는 포도당 이동과정의 변화로 FDG 섭취가 감소되었다. 또한 Pgp가 점차 증가됨과 함께 plasma membrane transporter인 GLUT-1 level이 감소된다. 이러한 결과는 다약제내성의 영상화가 지금까지의 예상보다 보다 복합적이고 다양하므로 보다 많은 연구가 필요할 것이라는 점을 시사한다. 최근 시도되고 있는 생체광학 영상을 이용한 다약제내성 유전자 및 Pgp 발현 연구는 아직 시작단계이나, 분자 생물학적 영상법의 발전과 함께 MRI 기술등에도 이용될 수 있으므로 향후 많은 연구가 있을 것으로 기대된다.

• 대한수의학회지
• /
• 제36권4호
• /
• pp.783-794
• /
• 1996

이온운반계는 생체의 각기 다른 세포의 성장을 조절하는 성장조절인자들의 효과를 매개하는데 깊은 관련이 있는 것으로 보고되고 있다. 신장 근위세뇨관에서 솔변 연 $Na^+/H^+$ 상호운반계는 사구체에서 여과된 나트륨의 재흡수와 수소이온의 분비를 조절하는 중요한 기능을 수행한다. 이 연구는 초대배양된 신장 근위세뇨관세포의 나트륨 운반을 Insulin-like Growth Factor-I(IGF-I)이 어떤 경로를 통하여 조절하는지를 알아보고자 실시하였다. 결과는 아래와 같다. 1. 초대배양된 신장 근위세뇨관세포에서 $Na^+$ uptake는 시간의존적으로 증가되었으며, 30분동안 $Na^+$ uptake를 실시한 결과 세포외 NaCl 농도의존적으로 $Na^+$ uptake를 유의성있게 감소시켰다(대조군; $40.11{\pm}1.76$, 140mM군; $17.82{\pm}0.94pmole\;Na^+/mg\;protein/min$). 2. $Na^+$ uptake는 iodoacetic acid(IAA, $1{\times}10^{-4}M$) 또는 valinomycin($5{\times}10^{-6}M$)처리시 대조군에 비해 각각 $50.51{\pm}4.4%$와 $57.65{\pm}2.27%$ 억제되었으며, ouabain($5{\times}10^{-5}M$)을 처리한 경우는 $140.23{\pm}3.37%$ 증가되었다. IGF-I($1{\times}10^{-5}M$)으로 배양한 세포를 actinomycin D($1{\times}10^{-7}M$)와 cycloheximide($4{\times}10^{-5}M$)로 처리시 $Na^+$ uptake는 대조군에 비해 각각 $90.21{\pm}2.39%$와 $89.64{\pm}3.69%$로 감소되었다. 3. IGF-I으로 배양한 세포에서 세포외 cAMP는 농도의존적($10^{-8}-10^{-4}M$)으로 $Na^+$ uptake를 유의성있게 감소시켰고, 3-isobutyl-1-methyl-xanthine(IBMX, $5{\times}10^{-5}M$)도 억제시켰다. Pertussis toxin(PTX, 50pg/ml)이나 cholera toxin(CTX, $1{\mu}g/ml$)의 처리시에도 $Na^+$ uptake는 억제되었다. 세포외 phorbol 12-myristate 13 acetate(PMA) 또한 농도의존적(1-100ng/ml)으로 $Na^+$ uptake를 감소시켰다. 그러나 staurosporine($1{\times}10^{-7}M$)은 $Na^+$ uptake에 영향을 미치지 않았으며 PMA와 stauiosporine을 동시에 처리했을 때도 $Na^+$ uptake는 억제되지 않았다. 결론적으로 초대배양된 토끼 신장 근위세뇨관세포에서 $Na^+$ uptake는 막전위와 세포내 에너지 의존적이며 IGF-I은 부분적으로 단백질 및 RNA 합성을 통해서 그리고 세포내 cAMP나 PKC 경로를 통해서 $Na^+$ uptake를 조절하는 것으로 생각된다.