• Journal of Chest Surgery
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• 제38권5호
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• pp.323-334
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• 2005

배경: 유전자 치료에 의한 치료적 혈관조성은 허혈성 심질환의 새로운 치료전략의 하나로 최근 많은 연구가 진행되고 있다. 본 연구의 목적은 대동물에서 pCK 플라스미드 벡터에 혈관내피성장인자(vascular endothelial growth factor isoform 165: VEGF165) 유전자를 삽입한 pCK-VEGF를 이용한 치료적 혈관조성의 효용성을 증명하는 것이다. 대상 및 방법: 총 21 마리의 돼지를 이용하여 좌전하행지동맥의 원위부를 결찰하여 심근경색 모델을 만든 후, 4주 후에 VEGF 유전자를 삽입한 플라스미드를 심근내에 주입하거나(VEGF군), 유전자 없이 플라스미드 만을 주입하였다(대조군). 실험 대상 동물군을 맹검하에 무작위로 VEGF군 및 대조군으로 나누어 실험을 진행하였는데, 7마리는 실험 도중 사망하였으며 결과적으로 VEGF군은 8마리, 대조군은 6마리가 최종분석에 이용되었다. 좌전하행지동맥 결찰 후 30일째에 심근 SPECT와 심장초음파검사를 시행하고 심근내에 플라스미드를 주입하였으며, 이로부터 30일째에 심근 SPECT와 심장초음파검사를 다시 시행하였다. 허혈부위의 심근관류의 변화는 심근 SPECT상의 $^{99m}Tc-MIBI$의 섭취 정도로 비교하였으며, 국소 및 전체 심근기능 및 심실리모델링 등은 심장초음파 또는 게이트SPECT 검사상의 수축시 심실벽비후화, 좌심실구출률(EF), 수축기말용적(ESV), 이완기말용적(EDV) 등으로 비교하였다. 혈관조성의 정도는 조직검사상의 미세혈관의 밀도를 측정하여 비교하였다. 결과: 미세혈관의 밀도는 VEGF군에서 유의하게 더 높았으며($386\pm110/mm^{2}\;vs.\;291\pm127/mm^{2},\;p<0.001$), 분절의 관류 정도도 VEGF군에서는 관상동맥 결찰 60일째가 30일째에 비해 더 증가한 반면(플라스미드 주입 전, 후, $48.4\pm15.2\%\;vs.\;53.8\pm19.6\%,\;p<0.001$) 대조군에서는 유의한 변화가 없었고(플라스미드 주입 전, 후, $45.1\pm17.0\%\;vs.\;43.4\pm17.7\%,\;p=0.186$), 그 변화량도 두 군간에 유의한 차이를 보였다($11.4\pm27.0\%$ 증가 vs $2.7\pm19.0\%$ 감소, p=0.003). 수축시의 심실벽비후화는 양 군 모두에서 플라스미드 주입 후 유의하게 증가하였으나 증가한 정도는 두 군간에 차이가 없었다. 심장초음파검사상 ESV은 양 군 모두에서 수술 전에 비해 관상동맥 결찰 후 유의하게 증가하였고 (VEGF군, $22.9\pm9.9\;mL\;vs.\;32.3\pm9.1\;mL,\;p=0.006;$ 대조군, $26.3\pm12.0\;mL\;vs.\;36.8\pm9.7\;mL,\;p=0.046$), EF은 유의하게 감소하였으며(VEGF군, $52.0\pm7.9\%\;vs\;46.5\pm7.4\%$, p=0.004; 대조군, $48.2\pm9.2\%\;vs\;41.6\pm10.0\%$, p=0.028), EDV은 양 군 모두에서 유의한 변화가 없었다. 플라스미드 주입 전과 후의 비교에서는 양 군 모두에서 심장초음파 및 게이트 SPECT검사상의 EF, ESV, EDV 값의 유의한 차이가 없었다. 결론: VEGF165 유전자를 삽입한 플라스미드의 심근내 주입 후 허혈성 생존 심근 부위에 혈관조성이 일어나고 심근관류가 유의하게 증가하였다. 그러나 심근 기능이나 좌심실의 리모델링 경과엔 유의한 차이가 없었다.

• BMB Reports
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• 제53권2호
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• pp.118-123
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• 2020

Cardiac regeneration with adult stem-cell (ASC) therapy is a promising field to address advanced cardiovascular diseases. In addition, extracellular vesicles (EVs) from ASCs have been implicated in acting as paracrine factors to improve cardiac functions in ASC therapy. In our work, we isolated human cardiac mesenchymal stromal cells (h-CMSCs) by means of three-dimensional organ culture (3D culture) during ex vivo expansion of cardiac tissue, to compare the functional efficacy with human bone-marrow derived mesenchymal stem cells (h-BM-MSCs), one of the actively studied ASCs. We characterized the h-CMSCs as CD90^low, c-kit^negative, CD105^positive phenotype and these cells express NANOG, SOX2, and GATA4. To identify the more effective type of EVs for angiogenesis among the different sources of ASCs, we isolated EVs which were derived from CMSCs with either normoxic or hypoxic condition and BM-MSCs. Our in vitro tube-formation results demonstrated that the angiogenic effects of EVs from hypoxia-treated CMSCs (CMSC-Hpx EVs) were greater than the well-known effects of EVs from BM-MSCs (BM-MSC EVs), and these were even comparable to human vascular endothelial growth factor (hVEGF), a potent angiogenic factor. Therefore, we present here that CD90^lowc-kit^negativeCD105^positive CMSCs under hypoxic conditions secrete functionally superior EVs for in vitro angiogenesis. Our findings will allow more insights on understanding myocardial repair.

• Journal of Ginseng Research
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• 제42권3호
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• pp.288-297
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• 2018

Background: The incidence of colorectal cancer (CRC) is increasing, with metastasis of newly diagnosed CRC reported in a large proportion of patients. However, the effect of Korean Red Ginseng extracts (KRGE) on epithelial to mesenchymal transition (EMT) in CRC is unknown. Therefore, we examined the mechanisms by which KRGE regulates EMT of CRC in hypoxic conditions. Methods: Human CRC cell lines HT29 and HCT116 were incubated under hypoxic (1% oxygen) and normoxic (21% oxygen) conditions. Western blot analysis and real-time PCR were used to evaluate the expression of EMT markers in the presence of KRGE. Furthermore, we performed scratched wound healing, transwell migration, and invasion assays to monitor whether KRGE affects migratory and invasive abilities of CRC cells under hypoxic conditions. Results: KRGE-treated HT29 and HCT116 cells displayed attenuated vascular endothelial growth factor (VEGF) mRNA levels and hypoxia-inducible $factor-1{\alpha}$ ($HIF-1{\alpha}$) protein expression under hypoxic conditions. KRGE repressed Snail, Slug, and Twist mRNA expression and integrin ${\alpha}V{\beta}6$ protein levels. Furthermore, hypoxia-repressed E-cadherin was restored in KRGE-treated cells; KRGE blocked the invasion and migration of colon cancer cells by repressing $NF-{\kappa}B$ and ERK1/2 pathways in hypoxia. Conclusions: KRGE inhibits hypoxia-induced EMT by repressing $NF-{\kappa}B$ and ERK1/2 pathways in colon cancer cells.

• Journal of Ginseng Research
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• 제46권6호
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• pp.771-779
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• 2022

Background: As a kind of common complication of the surgery of perianal diseases, perianal ulcer is known as a nuisance. This study aims to develop a kind of 20(S)-ginsenoside Rg3 (Rg3)-loaded hydrogel to treat perianal ulcers in a rat model. Methods: The copolymers PLGA₁₆₀₀-PEG₁₀₀₀-PLGA₁₆₀₀ were synthesized by ring-opening polymerization process and Rg3-loaded hydrogel was then developed. The perianal ulcer rat model was established to analyze the treatment efficacy of Rg3-loaded hydrogel for ulceration healing for 15 days. The animals were divided into control group, hydrogel group, free Rg3 group, Rg3-loaded hydrogel group, and Lidocaine Gel® group. The residual wound area rate was calculated and the blood concentrations of interleukin-1 (IL-1), interleukin-6 (IL-6), and vascular endothelial growth factor (VEGF) were recorded. Hematoxylin and eosin (H&E) staining, Masson's Trichrome (MT) staining, and tumor necrosis factor α (TNF-α), Ki-67, CD31, ERK1/2, and NF-κB immunohistochemical staining were performed. Results: The biodegradable and biocompatible hydrogel carries a homogenous interactive porous structure with 10 ㎛ pore size and five weeks in vivo degradation time. The loaded Rg3 can be released sustainably. The in vitro cytotoxicity study showed that the hydrogel had no effect on survival rate of murine skin fibroblasts L929. The Rg3-loaded hydrogel can facilitate perianal ulcer healing by inhibiting local and systematic inflammatory responses, swelling the proliferation of nuclear cells, collagen deposition, and vascularization, and activating ERK signal pathway. Conclusion: The Rg3-loaded hydrogel shows the best treatment efficacy of perianal ulcer and may be a candidate for perianal ulcer treatment.

• 한국응용곤충학회지
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• 제50권3호
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• pp.227-233
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• 2011

비단벌레(C. fulgidissima)는 동아시아에서 중풍을 치료하는 약으로 한국에서는 살충, 지양제로서 사용한 기록이 있다. 본 연구는 비단벌레의 에탄올 추출물의 내피세포에서의 산화질소(NO) 증강효과와 내피성 산화질소 합성효소(eNOS)의 양적 증가를 조사하였다. 그 결과 비단벌레 에탄올 추출물은 양성대조약물 sodium nitroprusside에 비해 65.9%의 NO 증강 효과를 가지는 것을 확인하였다. 또한 eNOS에 대해서도 농도 의존적으로 증가시킴을 확인하였다. 염증성부착인자인 ICAM-1과 VCAM-1 수치와 염증매개인자 프로스타글란딘 $E_2$를 조사하여 비단벌레 에탄올 추출물의 염증억제 기전을 조사한 결과, HUVEC 세포에서 농도 의존적으로 염증매개인자와 염증성부착인자의 감소를 확인하였다. 아울러 혈관성 내피성장인자(VEGF)의 낮은 수치를 HUVEC 세포에서 관찰할 수 있었다. 에탄올 추출물을 HPLC로 부분정제 후 GC-MS와 MALDI-TOF 분석을 통하여 일부 칸다리딘 성분 함유함을 확인하였다

• 대한한방내과학회지
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• 제35권4호
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• pp.416-427
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• 2014

Objectives: The purpose of this study was to investigate the antineoplastic effect of several herbal medicine mixtures (compositions of Astragalus membranaceu, Angelica gigas, Trichosanthes kirilowii, Panax ginseng, Rhus verniciflua Stokes) on the SNU-80 anaplastic thyroid carcinoma cell line. Methods: MTT assay was used to examine whether our herbal medicine mixtures decreased cell growth rate of SNU-80. Wound healing assay and Transwell invasion assay was performed to investigate whether our herbal medicine mixtures affect the migration and invasion of anaplastic cancer cells, SNU-80. ELISA assay was performed to know if our herbal medicine mixtures suppressed the expression of pro-invasive molecules, such as vascular endothelial growth factor (VEGF) and matrix metalloproteinase-2 (MMP-2) secreted from SNU-80. Results: MTT assay demonstrated that A. membranaceus:A. gigas:T. kirilowii=1:1:1 or 3:1:1, A. membranaceus:A. gigas :T. kirilowii:P. ginseng=1:1:1:1 or 3:1:1:1, A. membranaceus:A. gigas:T. kirilowii:P. ginseng:R. verniciflua Stokes=1:1:1:1:1 or 3:1:1:1:1 strongly suppressed the growth of SNU-80. Wound healing assay demonstrated that A. membranaceus:A. gigas=3:1, A. membranaceus:A. gigas:T. kirilowii=1:1:1 or 3:1:1, A. membranaceus:A. gigas:T. kirilowii:P. ginseng=1:1:1:1 or 3:1:1:1, A. membranaceus:A. gigas:T. kirilowii:P. ginseng:R. verniciflua Stokes=1:1:1:1:1 or 3:1:1:1:1 inhibited the migration of SNU-80. Transwell invasion assay demonstrated that A. membranaceus:A. gigas=1:1, A. membranaceus:A. gigas:T. kirilowii =1:1:1 or 3:1:1, A. membranaceus:A. gigas:T. kirilowii:P. ginseng=1:1:1:1, A. membranaceus:A. gigas:T. kirilowii:P. ginseng :R. verniciflua Stokes=1:1:1:1:1 or 3:1:1:1:1 inhibited the invasion of SNU-80. ELISA assay demonstrated that A. membranaceus :A. gigas:T. kirilowii=1:1:1 or 3:1:1, A. membranaceus:A. gigas:T. kirilowii:P. ginseng:R. verniciflua Stokes=1:1:1:1:1 suppressed the expression of VEGF. Also, A. membranaceus:A. gigas=1:1, A. membranaceus:A. gigas:T. kirilowii=1:1:1 or 3:1:1, A. membranaceus :A. gigas:T. kirilowii:P. ginseng=1:1:1:1 or 3:1:1:1, A. membranaceus:A. gigas:T. kirilowii:P. ginseng:R. verniciflua Stokes =1:1:1:1:1 or 3:1:1:1:1 suppressed the expression of MMP-2. Conclusions: The results obtained in this study suggest that several herbal medicine mixtures suppresse the growth and inhibit the migration and invasion of SNU-80, which is anaplastic thyroid cancer cells. Especially, A. membranaceus:A. gigas: T. kirilowii=1:1:1 mixture had a stronger anti-cancer effect.

• Archives of Plastic Surgery
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• 제38권6호
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• pp.733-739
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• 2011

Purpose: Cellulose is a natural substance from plants or bacteria. It is known that bacterial synthesized cellulose has an effect of wound healing. The aim of this study is to show the effect of bacterial synthesized cellulose from citrus on wound healing. Methods: Three full-thickness skin defects were made on the back of Sprague-Dawley rats. Three wounds were treated by vaseline gauze (Group V), Algisite $M^{(R)}$ (Group A) and bacterial synthesized cellulose from citrus (Group C) was used for dressing on skin defect on rats. We analyzed the gross, histological and biochemistry finding. Results: Group C showed more decrease of wound size compared to Group V (33% versus 7.2%) after 14 days. The histologic findings revealed Group C and Group A preceed the process of wound healing rather than Group V (More rapid collagen deposition and neovascularization and reduced inflammation). Also, the expressions of vascular endothelial growth factor (VEGF) and transforming growth factor (TGF)-${\beta}1$ were increased in the Group C and Group A compared with the Group V in 7 days. VEGF and TGF-${\beta}1$ expression were decreased in the Group C and Group A in 14 days, however Group V was not decreased at 14 day because of delayed wound healing process. Conclusion: Bacterial synthesized cellulose from citrus affects wound healing by reducing the inflammatory stage. And stimulates wound contracture by the deposition of extracellular matrix, thus preventing the formation of chronic wounds.

• Radiation Oncology Journal
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• 제34권3호
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• pp.223-229
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• 2016

Purpose: This study is to investigate the effect of captopril when combined with irradiation. Materials and Methods: 4T1 (mouse mammary carcinoma) cells were injected in the right hind leg of Balb/c mice. Mice were randomized to four groups; control (group 1), captopril-treated (group 2), irradiated (group 3), irradiated and captopril-treated concurrently (group 4). Captopril was administered by intraperitoneal injection (10 mg/kg) daily and irradiation was delivered on the tumor-bearing leg for 15 Gy in 3 fractions. Surface markers of splenic neutrophils (G-MDSCs) and intratumoral neutrophils (tumor-associated neutrophils [TANs]) were assessed using flow cytometry and expression of vascular endothelial growth factor (VEGF) and hypoxia-inducible factor 1 alpha ($HIF-1{\alpha}$) of tumor was evaluated by immunohistochemical (IHC) staining. Results: The mean tumor volumes (${\pm}$standard error) at the 15th day after randomization were $1,382.0({\pm}201.2)mm^3$ (group 1), $559.9({\pm}67.8)mm^3$ (group 3), and $370.5({\pm}48.1)mm^3$ (group 4), respectively. For G-MDSCs, irradiation reversed decreased expression of CD101 from tumor-bearing mice, and additional increase of CD101 expression was induced by captopril administration. Similar tendency was observed in TANs. The expression of tumor-necrosis factor-associated molecules, CD120 and CD137, are increased by irradiation in both G-MDSCs and TANs. Further increment was observed by captopril except CD120 in TANs. For IHC staining, VEGF and $HIF-1{\alpha}$ positivity in tumor cells were decreased when treated with captopril. Conclusion: Captopril is suggested to have additional effect when combined to irradiation in a murine tumor model by modulation of MDSCs and angiogenesis.

• Archives of Plastic Surgery
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• 제32권1호
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• pp.12-18
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• 2005

The Transverse rectus abdominis musculocutaneous (TRAM) flap has been commonly used for autologous breast reconstruction. Despite these clinical usefulness, the TRAM flap is prone to partial flap or fat necrosis in especially pedicled flap. To improve flap survival, the surgical delay procedures and pharmacological treatments have been developed. In many studies for the pharmacological treatment, Lipo-$PGE_1$ has demonstrated a marked ability to improve flap survival and it's effect has been proved similar to surgical delay procedure. The purpose of this study is to determine the most effective route of Lipo-$PGE_1$ administration as a pharmacological treatment in TRAM flap of the rat. Fifty male Sprague-Dawley rats weighing 300-350 gm were divided into five groups, One week before flap elevation, Lipo-$PGE_1$($2{\mu}g/kg$) was injected three times in a week and than the left inferior epigastric vessel based TRAM flap ($5.0{\times}3.0cm$) elevated; group I: no procedure before flap elevation; group II: intraperitoneal injection; group III: intravenous injection; group IV: subcutaneous injection; group V: topical application. A flap was assessed at postoperative 7 days by comparison of flap survival rate, vessel counts(H-E stain), and vascular endothelial growth factor(VEGF) protein expressed by Western blot. The results demonstrated that the mean percentages of the flap survival area in group III were significantly higher than that of any other group(p<0.05). The vessel counts of all experimental groups were statistically higher than that of control group(p<0.05). Only in group III, the VEGF protein expression was increased significantly than control group and there are no difference in other experimental groups. In conclusion, the intravenous administration of the Lipo-$PGE_1$ is the most effective on flap survival, and the VEGF induced by Lipo-$PGE_1$ has some positive effects on new vessel formation and flap survival.

• 대한본초학회지
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• 제28권6호
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• pp.101-109
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• 2013

Objectives : The purpose of this study was to investigate the effects of Polygoni Multiflori Radix Water Extract (PM) on the production of inflammatory mediators in RAW 264.7 mouse macrophages induced by lipopolysaccharide (LPS). Method : We examined effect of PM Extract on the cell viability of RAW 264.7 cells. Futhermore, we investigated anti-inflammatory effect of PM Extract by the production of proinflammatory cytokines such as NO, intracellular calcium, interleukin(IL)-$1{\alpha}$, IL-3, IL-$1{\beta}$, IL-6, interferon inducible protein-10(IP-10), keratinocyte-derived chemokine(KC) and vascular endothelial growth factor(VEGF). Result : No significant changes have been found in the mouse macrophge cell viability by the PM Extract at the concentration of 25, 50, 100 and $200{\mu}g/mL$. The water extract of PM significantly inhibited the production of NO and intracellular calcium in the LPS-induced macrophages at the concentration of 25, 50, 100 and $200{\mu}g/mL$. The water extract of PM significantly inhibited the production of IL-$1{\alpha}$, IL-${\beta}$, IL-3, IP-10, KC, VEGF in the LPS-induced macrophages at the concentration of 50, 100, and $200{\mu}g/mL$; IL-6 at the concentration of 100 and $200{\mu}g/mL$ ; and IL-17 at $200{\mu}g/mL$. Conclusion : The water extract of PM significantly inhibited the production of NO, intracellular calcium, IL-$1{\alpha}$, IL-3, IL-${\beta}$, IP-10, KC, VEGF at the concentration of 50 ㎍/mL or higher in the LPS-induced macrophages with no changes in the cell viability of them. These results suggest that water extract of Polygoni Multiflori Radix has anti-inflammatory effect regulating the production of proinflammatory cytokines in the LPS-induced macrophages.