• 한국안광학회지
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• 제9권2호
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• pp.333-343
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• 2004

Impression cytology는 결막상피의 표층을 떼어내기 위해 안구 표면에 cellulose acetate filter 재질을 같이 이용한다. 이 방법은 비 침습적이고 쉽게 시행할 수 있으며 환자에게 최소한의 불편감을 주며 결막표면에 반복해서 시행하여 변화를 관찰하기 위해 사용될 수 있다. 이 방법으로 결막 표면의 형태학적 연구와 squmaous metaplasia의 정도를 평가할 수 있다. 본 연구는 건성안 환자에서 impression cytology를 이용하여 결막표면을 평가하기 위해 수행되었다. 70명의 콘택트렌즈 비착용 학생을 대상으로 조사하였는데 먼저 조사 대상자들에게 McMonnies dry eye symptom questionnaire을 작성하도록 하였다. 그리고 비침습성 tear thinning time(TTT) 검사를 시행하고 다음에 침습성 검사인 Schirmer tear test(SIT), tear film break-up time(TBUT) tests 그리고 Rose-bengal staining올 시행하여 건성안 의심자를 분류하였다. 하 안구결막으로부터 결막 상피세포는 impression cytology기법으로 수집하여 PAS(Periodic Acid Schift)-haematoxylin 염색을 시행하였다. 배상세포와 결막상피세포는 광학현미경 400배에서 관찰하여 squamous metaplasia의 정도에 근거한 Nelson Grading scale에 따라 분류하였는데 이는 배상세포 밀도, 세포의 크기와 형태, 핵:세포질 비 등을 포함한다. 건성안 의심자는 상피세포의 형태적 변화, 핵의 변화, 배상세포 밀도 감소를 보였다. 세포학적 변화 정도는 건성안 조건의 정도에 관련이 있었다. 상피세포 형태를 Nelson의 분류시스템으로 단계를 나눈 결과 대조군의 91.43%가 grade 0 이었고 8.57%가 약간의 이상형태인 grade 1이었다. 반면에 건성안 의심자에서는 20%가 grade 0, 42.86%가 grade 1, 34.29%가 grade 2 그리고 2.86%가 심한 형태적 변화를 보인 grade 3를 나타냈다. Impression cytology는 부작용이나 거부반응이 없는 안구 표면 상피의 비 침습적 또는 최소 침습적 생검법으로 생각되며 안구표면을 포함하는 광범위한 과정을 위한 유용한 진단 기법으로 안전하고 간단하며 건성안 의심자의 다양한 안구 표면 변화의 이해를 증진시키는데 도움을 줄 수 있을 것으로 사료된다.

• 동의생리병리학회지
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• 제22권6호
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• pp.1500-1508
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• 2008

The author investigated whether Chwiyeon-tang(PC), Haengso-tang(PH), Jawanchihyo-san(PJ) and Gamisocheongryong-tang(PS) significantly affect both PMA-induced mucin production and MUC5AC gene expression in airway epithelial cells and sulfur-dioxide-induced airway goblet cell hyperplasia and mucus hypersecretion animal model using rat. Possible cytotoxicity of each herbal medicine was assessed by measuring the survival and proliferation rate of NCI-H292 cells. Confluent NCI-H292 cells were pretreated for 30 min in the presence of PC, PH, PJ and PS, respectively, and treated with PMA(10 $ng/m{\ell}$), to assess the effect of each herbal medicine on PMA-induced mucin production by enzyme-linked immunosorbent assay(ELISA). Effects of each herbal medicine on PMA-induced MUC5AC gene expression from the same cells were investigated. Also, hypersecretion of airway mucus and goblet cell hyperplasia were induced by exposure of rats to $SO_2$ during 3 weeks. Effects of orally-administered PC, PH, PJ and PS during 1 week on intraepithelial mucosubstances and hyperplasia of goblet cells were examined using histological analysis after staining the epithelial tissue with PAS-alcian blue. (1) PC, PJ, PS and PH did not show significant effects on the survival and proliferation of NCI-H292 cells ; (2) PC, PJ and PS significantly decreased PMA-induced mucin production from NCI-H292 cells ; (3) PC, PJ and PS significantly inhibit the expression levels of PMA-induced MUC5AC gene in NCI-H292 cells ; (4) Among PC, PJ, PS and PH, only PS decreased $SO_2$-induced hyperplasia of airway goblet cells and intraepithelial mucosubstances. This result suggests that PC, PJ and PS can not only affect the production of mucin but also affect the expression of mucin gene and this can explain, at least in part, the traditional use of PC, PJ and PS for controlling airway diseases showing hypersecretion of mucus in oriental medicine.

• 대한한방소아과학회지
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• 제29권3호
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• pp.65-79
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• 2015

Objectives : In this study, effects of haepyoijintang (HIJ) on the increase in airway epithelial mucosubstances of rats and ATP-, PMA-, EGF- or TNF-${\alpha}$-induced MUC5AC mucin production and gene expression from human airway epithelial cells were investigated. Methods : Hypersecretion of airway mucus was induced by exposure of rats to $SO_2$ during 3 weeks. Effect of orally-administered HIJ during 2 weeks on increase in airway epithelial mucosubstances from tracheal goblet cells of rats was evaluated using histopathological analysis after staining the epithelial tissue with PAS-alcian blue. Possible cytotoxicity of HIJ was evaluated by examining the potential damage of kidney and liver functions by measuring serum GOT/GPT activities and serum BUN and creatinine concentrations of rats and the body weight gain during experiment, after administering HIJ orally. At the same time, the effect of HIJ on ATP-, PMA-, EGF- or TNF-${\alpha}$-induced MUC5AC mucin production and gene expression from human airway epithelial cells (NCI-H292) were investigated. Confluent NCI-H292 cells were pretreated for 30 min in the presence of HIJ and treated with ATP ($200{\mu}M$), PMA (10 ng/ml), EGF (25 ng/ml) or TNF-${\alpha}$ (0.2 nM) for 24 hrs, to evaluate the effect of HIJ both on ATP-, PMA-, EGF- or TNF-${\alpha}$-induced MUC5AC mucin production using enzyme-linked immunosorbent assay (ELISA) and on gene expression by the same inducers using reverse transcription-polymerase chain reaction (RT-PCR). Results : (1) HIJ decreased the amount of intraepithelial mucosubstances of trachea of rats. (2) HIJ did not show renal and hepatic toxicities and did not affect body weight gain of rats during experiment. (3) HIJ significantly inhibited ATP-, PMA-, EGF-, and TNF-${\alpha}$-induced MUC5AC mucin productions from NCI-H292 cells. (4) HIJ significantly inhibited ATP-, PMA-, EGF-, and TNF-${\alpha}$-induced MUC5AC mucin gene expression from NCI-H292 cells. Conclusions : The result from the present study suggests that HIJ might control the production and gene expression of airway mucin observed in various respiratory diseases accompanied by mucus hypersecretion and do not show in vivo toxicity to liver and kidney functions after oral administration. Effect of HIJ with their diverse components should be further investigated using animal experimental models that can reflect the pathophysiology of airway diseases through future studies.

• 대한치과보철학회지
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• 제37권5호
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• pp.698-713
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• 1999

Adherence of Candida albicans(C. albicans) to the surface of a denture is believed to be an initial and essential step in the formation of denture-induced stematitis. Previous studies have provided enormous infomation on the relationship between composition of palatine gland/parotid saliva and upper denture stomatitis. Relatively little information is available on the correlation between lower denture stomatitis and sublingual-submandibular ( SLSM ) saliva. The plaque samples were collected from the two sites($100mm^2$) on the inner surface of lower partial denture corresponding to the stematitis and healthy region of the lower partial dentures of 12 denture stomatitis patients and 6 nor-mal persons who wore lower partial dentures. The samples were plated to isolate C. albicans on a selective Saboraud's dextrose agar plate and the isolates were identified by germ tube test and gram staining. The subjects were divided into group I (stomatitis with C. albican), group II (lesion without C. albicans), group III (no lesion but C. albicans), and group IV (normal and healthy denture wearer). Individual SLSM saliva($20{\mu}g$ of protein) was analyzed by SDS-PAGE (SDS -poly-acrylamide gel electrophoresis) with Coomassie brilliant blue and PAS(Periodic Acid Schinff) stain-ing. The salivary proteins separated in the polyacryamide gels were subjected to immunoblot anaysis using anti-lactoferrin, anti-sIgA, and anti-secretory component of sIgA. In this study using custom made acrylic denture resin beads(5mm in diameter) coated with stimulated individual SLSM saliva, the binding ability of individual C. albicans strains to the beads was observed. Levels of C, albicans adhered to the acrylic resin beads were determined by measuring the optical density of the bound C. albicans to the beads at 580nm. The results showed that a higher number of C. albicans was observed in the lesion site than healthy site. The saliva of group I contained more high molecular weight glycoprotein(mucin, MGI) as compared to group II, III and IV. And lactoferrin and sIgA affected to the binding ability of C. albicans to acylic resin beads. Binding ability of individual C. albicans to the acrylic resin coated with respective individual saliva was found to be greater in group I than the other 3 groups. And when bound cells of C. albicans isolated from individual subject #2 to the saliva coated beads were used binding ability of subject #2 saliva coated beads was founed to be greater than the other sutjects. These results suggested that denture induced stomatitis is related to individual patient's salivary protein composition, especially MG-1. Future studies will be directed toward saliva exam-ination of patients who have general disease and analysis of pellicles formed on prosthesis with respect to oral disease.

• 대한한방소아과학회지
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• 제27권1호
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• pp.69-81
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• 2013

Objectives In this study, effects of Macmundongtang (MMT) on ATP or TNF-${\alpha}$ or PMA or EGF induced MUC5AC mucin production and gene expression from human airway epithelial cells and the increase in airway epithelial mucosubstances of rats were investigated. Materials and Methods Confluent NCI-H292 cells were pretreated for 30min in the presence of MMT and treated with ATP ($200{\mu}M$) or PMA (10 ng/ml) or EGF (25 ng/ml) or TNF-${\alpha}$ (0.2 nM) for 24hrs, to assess the effect of MMT both on ATP- or PMA- or EGF- or TNF-${\alpha}$-induced MUC5AC mucin production using enzyme-linked immunosorbent assay (ELISA) and on gene expression by the same inducers using reverse transcription-polymerase chain reaction (RT-PCR). At the same time, hypersecretion of airway mucus was induced by exposure of rats to SO2 during 3 weeks. Effect of orally-administered MMT during 2 weeks on increase in airway epithelial mucosubstances from tracheal goblet cells of rats was assesed using histopathological analysis after staining the epithelial tissue with PAS-alcian blue. Possible cytotoxicity of MMT was assessed by investigating the potential damage of kidney and liver functions by measuring serum GOT/GPT activities and serum BUN concentration of rats and the body weight gain during experiment, after administering MMT orally. Results (1) MMT did not only inhibit but also increased MUC5AC mucin productions and expression levels of MUC5AC gene from NCI-H292 cells. (2) MMT did not decrease the amount of intraepithelial mucosubstances of trachea of rats. (3) MMT did not show renal and hepatic toxicities and did not affect body weight gain of rats during experiment. Conclusions The result from the present study suggests that MMT might normalize the production and gene expression of airway mucin observed in various respiratory diseases accompanied by yin-deficiency, without in vivo toxicity to liver and kidney functions after oral administration.

• 대한한방소아과학회지
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• 제30권2호
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• pp.31-46
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• 2016

Objectives In this study, the effects of Ja-eum-gang-hwa-tang (JGT) on the increase in airway epithelial mucosubstances of rats and ATP- or PMA- or EGF- or TNF-${\alpha}$-induced MUC5AC mucin production and gene expression from human airway epithelial cells were investigated. Materials and Methods Hypersecretion of airway mucus was produced by exposure of $SO_2$ to rats for 3 weeks. The effect of orally-administered JGT for 2 weeks on increased epithelial mucosubstances from tracheal goblet cells of rats was assessed by using histopathological analysis after staining the epithelial tissue with Hematoxylin-eosin and PAS-alcian blue. Possible cytotoxicity of JGT was assessed by investigating the potential damage on kidneys and liver functions by measuring serum GOT/GPT activities and serum BUN concentration of rats and the body weight gain during experiment. Also, the effect of JGT on ATP- or PMA- or EGF- or TNF-${\alpha}$-induced MUC5AC mucin production and gene expression from human airway epithelial cells (NCI-H292) were investigated. Confluent NCI-H292 cells were pretreated for 30 min in the presence of JGT and treated with ATP ($200{\mu}M$) or PMA ($10ng/ml$) or EGF ($25ng/ml$) or TNF-${\alpha}$ (0.2 nM) for 24 hrs to assess the effect of JGT both on ATP- or PMA- or EGF- or TNF-${\alpha}$-induced MUC5AC mucin production by using enzyme-linked immunosorbent assay (ELISA) and on gene expression by the same inducers using reverse transcription-polymerase chain reaction (RT-PCR). Results (1) JGT decreased the amount of intraepithelial mucosubstances of trachea of rats. (2) JGT did not show any renal and hepatic toxicities, and did not affect body weights either. (3) JGT significantly inhibited ATP-, PMA-, EGF-, and TNF-${\alpha}$-induced MUC5AC mucin productions from NCI-H292 cells. (4) JGT inhibited EGF-, and PMA-induced expression levels of MUC5AC gene in NCI-H292 cells. However, ATP- and TNF-${\alpha}$-induced MUC5AC gene expression levels were not affected in NCI-H292 cells. Conclusions The result from the present study suggests that JGT might control the production and gene expression of airway mucin observed in various respiratory diseases which accompanied by mucus hypersecretion. Also, JGT did not show liver toxicity or impact on kidney functions. The effect of JGT should be further studied by using animal experimental models which can show proper pathophysiology of airway diseases.

• 생명과학회지
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• 제27권4호
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• pp.450-455
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• 2017

돌더덕은 전통의학의 소재로서 기관지염, 기침, 경련, 부종, 간염, 대장염, 폐 손상의 치료로 사용되어 왔다. 돌더덕 추출물의 천식증상 완화를 연구하기 위하여, 먼저 항산화 활성을 비교한 결과, DPPH, FRAP, Cupper 소거활성 실험으로부터 농도 의존적으로 우수한 활성을 확인할 수 있었다. 오브알부민 유도의 마우스 천식 동물모델에 적용한 결과, histamine, IL-31, MMP-9의 발현이 현저하게 감소함을 알 수 있었다. 또한 IL-13, 면역세포수, 호산구의 발현이 73.7%, 73.5%, 80.9% 정도 억제되었다. 이의 결과로, 돌더덕 추출물은 오브알부민 유도의 마우스 천식 동물모델에서 탁월한 천식 효과를 나타내므로 항천식 식품소재나 바이오소재로의 개발이 기대된다.