In Vitro에서 TGF β1, α-SMA 유전자 발현을 통한 Copper tripeptide의 표피재생 효과

김극준,도우주,김정희 한국미용예술경영학회 2023 미용예술경영연구 Vol.17 No.6

This study aimed to investigate whether copper tripeptide, a cosmetic ingredient available on the market, has an epidermal regeneration effect in vitro. To confirm whether copper tripeptide has cytotoxicity, an MTT assay was performed, and the results showed no cytotoxicity at 100 μg/mL. Total RNA was extracted from fibroblasts (NIH3T3 cells) and epidermal keratinocytes (HaCaT cells) treated with copper tripeptide, and the mRNA expression levels of TGF β1 and α-SMA were examined using RT-PCR. The results confirmed that copper tripeptide increased the expression of TGF β1 and α-SMA genes in both epidermal keratinocytes and fibroblasts. To assess skin regeneration, a wound healing assay was used to examine cell growth and migration of NIH3T3 and HaCaT cells. The results showed that copper tripeptide had a cell regeneration-inducing effect compared to the control group in both NIH3T3 and HaCaT cells. These findings suggest that non-toxic copper tripeptide may have the potential as a cosmetic ingredient to enhance epidermal regeneration in NIH3T3 fibroblasts and HaCaT cells through increased TGF β1 and α-SMA gene expression and wound healing analysis.

밀웜 오일을 함유한 트리트먼트제의 손상모발 개선 효과

이혜경,김극준,김정희 한국미용예술경영학회 2023 미용예술경영연구 Vol.17 No.5

In this study, we analyzed the hair protective effect of treatment agent containing mealworm oil on hair damaged by bleaching. The sample hair was treated pre- and post-treatment with dose-dependent manner (0%, 3%, and 7% mealworm oil). Measured the absorbance via methylene blue staining to see the degree of damage to the hair, the absorbance value of VH was 0.24±0.02 (O.D) and BH was 2.64±0.20 (O.D), indicating that the absorbance value of bleached hair is higher than that of healthy hair. The absorbance value of 7TH with 7% of mealworm oil was 1.81±0.63 (O.D), which was 31.44% lower than that of the control group BH. In order to find out the physical characteristics of damaged hair, the maximum stress, tensile distance, and tensile strength of each hair were measured. As a result, the maximum stress increased by 7TH than VH, and the tensile distance increased as the contents of mealworm oil increased compared to BH. TH and 3TH showed similar tensile strength values to BH, but the tensile strength value of hair treated with 7TH increased. Statistical analysis showed that the standard deviation value for each sample was large and there was no significance in the average value. As a result of observing the morphologically damaged hair, the cuticle layer was dehiscence or desquamation in the hair damaged by bleaching compared to the virgin hair (VH). However, in the case of hair treated with mealworm oil, the gap in the cuticle layer are filled and a relatively regular layer can be seen. When treatment was used with mealworm oil added to damaged hair, there was improvement effect was shown on the cuticle and cortex. Therefore, based on this study, it is considered that mealworm oil is highly useful as a materials for hair products and cosmetics by proving that the fatty acid component of mealworm oil has an improvement effect on damaged hair.

포도전정가지 추출물이 UVB에 유도된 피부 손상에 대한 알레르기와 피부증식 억제효과

김정희,김대환,김극준 대한임상검사과학회 2017 대한임상검사과학회지(KJCLS) Vol.49 No.4

This study examined the effects of polyphenols in grape pruning stem extracts (GPSE) using grape stems discarded after harvest. The inhibitory effects on allergy, proliferation, and apoptosis in UVB-induced HR-1 hairless mice were analyzed. The applicability as a material for functional food and functional cosmetics was evaluated. The contents of the active ingredients of GPSE were analyzed by HPLC. After UVB irradiation on the dermis of HR-1 hairless mice, the number of mast cells was determined by toluidine blue staining to confirm that the skin allergic reaction was caused by GPSE. The results were similar to the normal group in the group applied GPSE, and there was no allergic reaction in the GPSE application group and a significant decreased compared to the sun cream control. PCNA immunohistochemical staining of the epidermal proliferation factor had an inhibitory effect on epidermal epithelial cell proliferation in all concentrations of GPSE and serum base mixture as an average of 42% compared to the control group. The mixture of GPSE and serum base suppressed the apoptosis inhibition rate by 27% on average compared to the control. These results confirmed the inhibitory effects GPSE on the allergic, proliferation, and apoptosis activities by with a serum base on UVB-induced skin damage. GPSE is a functional ingredient with potential skin protection effects, and has a high utilization as an ingredient for functional cosmetics. 본 연구는 포도수확 후 버려지는 가지를 이용한 포도전정가지 추출물(grape pruning stem extracts, GPSE)에 함유된polyphenol류가 UVB로 유도된 HR-1 hairless mice의 손상된피부에 대한 피부증식 억제, 항 알레르기, 아포토시스 억제능을분석하여 기능성 화장품 소재로서의 응용 가능성을 확인하고자하였다. 포도전정가지에서 polyphenol 성분을 80% 에탄올로추출하고 여과하여 농축한 후, 동결건조하여 −20°C에 보관하면서 사용하였다. GPSE의 유효성분 함량은 HPLC를 사용하여선행연구논문에 보고하였다. 피부손상을 유도하기 위해 UVB 를 HR-1 hairless mice에 조사하였고, GPSE가 피부 알레르기반응을 일으키는지 확인하기 위하여 toluidine blue 염색을 통하여 비만세포의 수를 확인한 결과 포도전정가지 추출물(GPSE)를 도포한 그룹에서는 정상군과 유사한 결과를 나타내어 GPSE 도포군은 알레르기 반응이 없으며 suncream 대조군대비 현저하게 감소하는 것을 확인할 수 있었다. 표피의 증식표지자인 PCNA 면역조직화학적 염색을 시행한 결과 GPSE와serum base 혼합물의 모든 농도에서 대조군 대비 평균 42%로표피상피세포 증식 억제 효과를 나타내었다. 또한 GPSE와serum base 혼합물이 자외선 조사로 인한 아포토시스 억제율은 대조군 대비 평균 27% 억제 한다는 것을 알 수 있었다. 이상의 결과로 포도전정가지로부터 추출한 페놀성 화합물이 화장품기초 원료인 serum base에 함유토록 제조된 GPSE는 피부증식억제, 알레르기 억제, 아포토시스 억제효과를 얻을 수 있었다. 본 연구의 의의는 지역포도농가에서 수확 후 버려지는 포도전정가지에서 폴리페놀류와 같은 유효성분을 확보하여 고부가가치를 창출할 수 있는 기능성화장품 원료로서의 활용 가치가 높을 것으로 기대된다.

면역조직화학적 염색 방법에 따른 상피세포 성장 수용체 단백(EGFR)의 발현정도의 차이 및 EGFR의 발현정도와 EGFR 유전자의 돌연변이와의 상관관계에 대한 고찰

윤인숙 ( In Sook Yoon ),김극준 ( Keuk Jun Kim ),이은화 ( Eun Hwa Lee ),석상희 ( Sang Hee Seok ),김상희 ( Sang Hee Kim ),김현용 ( Hyun Yong Kim ),송호정 ( Ho Jung Song ),이태종 ( Tae Jong Lee ) 대한임상검사과학회 2007 대한임상검사과학회지(KJCLS) Vol.39 No.3

In the last 5 years the Epidermal Growth Factor Receptor (EGFR) has emerged as one of the most important targets for drug development in oncology. Monoclonal antibodies targeting the external domain of EGFR have been shown to have clinical benefits in colorectal and head and neck cancer when combined with chemotherapy and/or radiation. Also the targeting of the epithelial growth factor receptor (EGFR) kinase domain using the closely related inhibitors gefitinib and erlotinib has generally been ineffective against solid tumors, many of which over express the receptor. We found that there were some differential expressions according to primary antibodies of the EGFR protein which being used as one of the histological tumor markers for non-small cell lung cancer (NSCLC). We also found that there are some differential expressions according to antibodies, the pH of the antigen retrieval (AR) buffer solutions and kinds of enzymes. There were some differential expressions according to the secondary antibodies and the detection systems. We analyzed the correlations between the immunohistochemical expressions of the EGFR protein and the gene mutations of the EGFR. The differences between automatic stainers and manual staining methods were also evaluated.

피부사상균의 항진균제 감수성 검사

문석기,신동훈,최종수,김기홍,김극준 대한의진균학회 2008 대한의진균학회지 Vol.13 No.2

Background: A standardized reference method for dermatophytes in vitro susceptibility testing is lacking. However, with increasing variety of drugs available to treat dermatophytosis, the need for a reference method for dermatophytes testing has become apparent. Objective: To evaluate a method of quantifying dermatophytes, the standards for broth microdilution method and evaluation of the availability of disk diffusion method in antifungal susceptibility testing for dermatophytes. Methods: 14 Candida species (sp.), 30 Trichophyton(T.) mentagrophytes, 9 T. raubitschekii and 11 T. rubrum were tested for fluconazole susceptibility by the broth microdilution method and disc diffusion method. Candida sp. was tested according to National Committee for Clinical Laboratory Standards (NCCLS) M27-A and M44-A. Broth microdilution method for T. mentagrophytes, T. raubitschekii and T. rubrum was operated according to NCCLS M38-A. Disk diffusion method for T. mentagrophytes, T. raubitschekii and T. rubrum was tested refer to NCCLS M44-A. Result: The disk diffusion method showed 50% correlation rate with the broth microdilution method for antifungal susceptibility testing for Candida species. The MIC reading point and incubation time of the broth microdilution method for T. mentagrophytes, T. raubitschekii and T. rubrum are Spec-50 and from 7, 6 and 9 days. Relation between the broth microdilution method and disk diffusion method for dermatophytes is poor. Conclusions: The good method for quantifying dermatophytes is using vortexing only or liquid nitrogen and homogenizer. Standards of MIC reading point and incubation time of microdilution method for dermatophytes are Spec-50 and from 6 to 9 days. It appears that the disk diffusion method is not recommended method for the antifungal susceptibility testing of dermatophytes. Background: A standardized reference method for dermatophytes in vitro susceptibility testing is lacking. However, with increasing variety of drugs available to treat dermatophytosis, the need for a reference method for dermatophytes testing has become apparent. Objective: To evaluate a method of quantifying dermatophytes, the standards for broth microdilution method and evaluation of the availability of disk diffusion method in antifungal susceptibility testing for dermatophytes. Methods: 14 Candida species (sp.), 30 Trichophyton(T.) mentagrophytes, 9 T. raubitschekii and 11 T. rubrum were tested for fluconazole susceptibility by the broth microdilution method and disc diffusion method. Candida sp. was tested according to National Committee for Clinical Laboratory Standards (NCCLS) M27-A and M44-A. Broth microdilution method for T. mentagrophytes, T. raubitschekii and T. rubrum was operated according to NCCLS M38-A. Disk diffusion method for T. mentagrophytes, T. raubitschekii and T. rubrum was tested refer to NCCLS M44-A. Result: The disk diffusion method showed 50% correlation rate with the broth microdilution method for antifungal susceptibility testing for Candida species. The MIC reading point and incubation time of the broth microdilution method for T. mentagrophytes, T. raubitschekii and T. rubrum are Spec-50 and from 7, 6 and 9 days. Relation between the broth microdilution method and disk diffusion method for dermatophytes is poor. Conclusions: The good method for quantifying dermatophytes is using vortexing only or liquid nitrogen and homogenizer. Standards of MIC reading point and incubation time of microdilution method for dermatophytes are Spec-50 and from 6 to 9 days. It appears that the disk diffusion method is not recommended method for the antifungal susceptibility testing of dermatophytes.

포도전정가지 추출물이 UVB로 유도된 결합 조직 손상에 미치는 피부 보호 효과

김정희(Joung-Hee Kim),김극준(Keuk-Jun Kim) 한국생명과학회 2018 생명과학회지 Vol.28 No.2

본 연구는 포도수확 후 버려지는 가지를 이용한 포도전정가지 추출물(Grape Pruning Stem Extracts, GPSE)에 함유된 polyphenol류 중 항산화, 항염증, 항암 등의 효과가 있다고 알려진 rutin, procyanidin B3, quercetin, kaempferol의 함량을 분석하고, UVB로 유도된 HR-1 mice의 손상된 피부에 대한 교원섬유, 탄력섬유 파괴억제 효과를 측정하여 기능성 화장품 소재로서의 응용 가능성을 확인하고자 하였다. 포도전정가지에서 polyphenol 성분을 80% EtOH로 추출하고 여과하여 농축한 후, 동결건조하여 -20℃에 보관하면서 사용하였다. GPSE의 유효성분 함량은 HPLC를 사용하여 분석하였다. 피부손상을 유도하기 위해 UVB를 HR-1 hairless mice에 조사하였고, 손상된 피부에 교원섬유 파괴억제 효과를 보기 위해 Masson’s trichrome stain, 탄력섬유 파괴 억제 효과를 보기 위해 Verhoeff’s elastic fiber stain, MMP-9 단백 발현을 보기 위해 면역조직화학적 염색을 실시하였다. 본 연구에 사용된 포도전정가지 시료 53 kg에서 EtOH 분획 추출물 2.34 kg을 추출하여 4.42%의 수율 결과를 얻었다. 유효성분 분석결과, procyanidin B3, 0.28 mg/g, rutin 12.81 mg/g, quercetin 0.51 mg/g 및 kaempferol 8.24 mg/g로 나타났다. GPSE와 serum base를 혼합 도포한 그룹에서 UVB에 의한 교원섬유와 탄력섬유를 분해하는 MMP-9 단백발현 결과, 대조군 대비 평균 약 90% 감소 억제하는 효과를 볼 수 있었다. 교원섬유 파괴억제 효과는 모든 농도에서 평균 24%의 억제율과, GPSE와 serum base의 혼합물 농도 2,000 mg/kg 에서 43%로 가장 우수한 교원섬유 파괴억제효과가 나타났으며, 탄력섬유파괴 억제 효과는 모든 농도에서 평균 12%의 억제율과 GPSE와 serum base의 혼합물 농도 1,000 mg/kg에서 35%로 가장 우수한 탄력섬유 파괴 억제 효과가 있는 것으로 나타났다. 이 결과는 GPSE가 진피층의 세포 외 기질에 UVB 조사에 의해 파괴되는 교원섬유, 탄력섬유의 파괴를 억제하는 작용을 한다는 것을 알 수 있었다. 이상의 결과로 포도전정가지로부터 rutin, kaempferol, quercetin, procyanidin B3 등의 폴리페놀성 화합물이 화장품 기초 원료인 serum base에 함유토록 제조된 GPSE 는 교원섬유 파괴 억제, 탄력섬유파괴 억제효과 및 피부손상 기능개선 효과의 결과를 얻을 수 있었다. 본 연구의 의의는 지역포도농가에서 수확 후 버려지는 포도전정가지에서 폴리페놀류와 같은 유효성분을 확보하여 화장품 원료로서의 활용 가치가 높을 것으로 기대된다. This study aimed to analyze the contents of rutin, procyanidin B3, quercetin, and kaempferol, known to have antioxidant, anti-inflammatory, and anti-carcinogenic effects, among the polyphenol types contained in grape pruning stem extracts (GPSE). It utilized grape stems discarded after harvest to measure the effects of GPSE on skin moisture, inhibition of skin cell proliferation, and anti-inflammatory activity on the damaged skin of HR-1 mice induced with ultraviolet B (UVB), and to verify the applicability of GPSE as a material for functional food and functional cosmetics. The polyphenol was extracted from grape pruning stems with 80% EtOH, and then the extract was used while storing at -20℃, after filtering, concentrating, and freeze-drying it. The content of an active ingredient of GPSE was analyzed using high performance liquid chromatography (HPLC). From 53 kg of the grape pruning stem specimen, 2.34 kg of the EtOH fraction extracts were extracted to achieve a 4.42% yield ratio. Analysis of the active ingredients showed 0.28 mg/g of procyanidin B3, 12.81 mg/g of rutin, 0.51 mg/g of quercetin, and 8.24 mg/g of kaempferol. After UVB irradiation on the dermis, to confirm the degree of inhibition of collagen synthesis, we examined the protein expression of MMP-9 using immunohistochemical staining. The results of this study confirm the existence of active polyphenol types, such as rutin, kaempferol, quercetin, and procyanidin B3, in GPSE. Moreover, the study found that GPSE has anti-collagenase effects and it decreases the effects of UV damage on skin barrier function. GPSE is a functional ingredient with a potential for skin protection effects, and it has high utilization potential as an ingredient for functional cosmetics.

조갑진균증의 진단에 있어 KOH 검사, 진균 배양 검사, 병리 조직 검사 및중합효소 연쇄반응의 비교

신영민,신동훈,최종수,김기홍,김극준 대한의진균학회 2007 대한의진균학회지 Vol.12 No.2

Background: Onychomycosis is one of the most common nail disorders, and it is necessary of mycologic confirmation to treat correctly. It has been reported that histopathologic evaluation and polymerase chain reaction (PCR) of the nail plate by nail clipping might be a useful supportive method for identification of causative fungi showing higher detection rates. Objective: This study was designed to compare diagnostic efficacy of KOH preparation, fungal culture, histopathologic examination and PCR in onychomycosis. Methods: Using 60 nail samples from 60 patients with clinically suspected onychomycosis, KOH preparation and fungal culture with nail and subungual tissue was performed. And histopathologic examination with PAS staining and PCR with DNA extracted from paraffin block was performed. Results: The results are summarized as follows: 1. The positive rates of conventional KOH preparation, fungal culture, histopathologic examination and PCR were 87.5%, 8.9%, 87.5% and 83.9%. 2. In histopathologic examination, fungi were distinguished as 3 groups based on Kim and Cho's classification (1997): dermatophytes, Candida sp. and non-dermatophytic mold (NDM). Fifteen of 49 samples are dermatophytes, 1 is Candida sp., and 7 are NDM, 8 are dermatophytes and Candida sp. mixed infections, 18 are dermatophytes and NDM mixed infections. 3. In PCR, 1 of 47 samples is Trichophyton rubrum, 4 are Candida sp. and 39 samples are NDM. Conclusion: Histopathologic examination in onychomycosis is considered to be an useful supportive method in diagnosis and identification of causative fungi in onychomycosis. [Kor J Med Mycol 2007; 12(2): 59-69]

Effect of Different Ingredients in Traditional Korean Medicine for Human Uterine Leiomyoma on Normal Myometrial and Leiomyomal Smooth Muscle Cell Proliferation

Prati Bajracharya,이은주,이동목,Sang Hee Shim,김극준,Sung Ho Lee,Jei Jun Bae,전상식,Tae Kyun Lee,Seok Hoon Kwon,최인호 대한약학회 2009 Archives of Pharmacal Research Vol.32 No.11

Crude water extracts of 13 traditional Korean medicinal ingredients used for leiomyomal treatment were prepared and used to treat human uterine normal myometrial and leiomyomal cell cultures. All the ingredients inhibited proliferation and altered the morphology of both myometrial and leiomyomal cells. Among the 13 ingredients, n-hexane-, chloroform-, and ethylacetate-soluble fractions were extracted from seven ingredients that potently inhibited cell proliferation in their water extract form. Among these, the ethylacetate-fraction of Phlomis umbrosa and Spatholobus suberectus, and the chloroform-fraction of Curcuma zedoaria and S. suberectus inhibited leiomyomal cell proliferation significantly compared to myometrial cell proliferation. Similarly, immunohistochemical analysis showed the inhibition of transforming growth factor-beta receptor 2 in leiomyomal tissue after treatment with the fractions of the ingredients. Moreover, the chloroform-fraction of C. zedoaria was subfractionated by open column chromatography. Two of the eight subfractions (fractions 6 and 7) potently inhibited cell proliferation in leiomyoma compared to myometrium. Further study will be performed with the goal of isolating specific compounds from two effective subfractions of C. zedoaria, ethylacetate-fraction of P. umbrosa, and the ethylacetate and chloroform-fractions of S. suberectus. The present study may be helpful in developing an alternative remedy to leiomyoma with minimal side-effects compared to the current treatments.

면역조직화학적 염색 방법에 따른 상피세포 성장 수용체 단백(EGFR)의 발현정도의 차이 및 EGFR의 발현정도와 EGFR 유전자의 돌연변이와의 상관관계에 대한 고찰

윤인숙,김상희,김현용,송호정,이태종,이은화,석상희,김극준 대한임상병리사협회 2007 조직세포검사학회 발표자료집 Vol.2007 No.-

Britanin attenuates ovalbumin-induced airway inflammation in a murine asthma model

김선건,이의진,박나영,박효현,정규태,김극준,이윤주,Meihua Jin,이은경 대한약학회 2016 Archives of Pharmacal Research Vol.39 No.7

We previously demonstrated the alleviation of ovalbumin (OVA)-induced airway inflammation by Inulae flos. In the present study, the effects of britanin, a sesquiterpene compound isolated from Inulae flos, were evaluated in an in vivo animal model for anti-asthma activity through observation of airway hyperresponsiveness (AHR), eosinophil recruitment, Th2 cytokine and IgE levels, and lung histopathology. Britanin administration effectively reduced AHR induced by aerosolized methacholine, airway eosinophilia, Th2 cytokines in bronchoalveolar lavage fluids and the supernatant of cultured splenocytes compared with OVA-induced mice. Histological studies showed that increased inflammatory cell infiltration and mucus secretion were reduced by britanin administration. Thus, britanin may have therapeutic potential for treating allergic asthma.