• Title/Summary/Keyword: Histological measurement

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Effect of suprascapular nerve injury on muscle and regenerated enthesis in a rat rotator cuff tear model

  • Kenichiro Eshima;Hiroki Ohzono;Masafumi Gotoh;Hisao Shimokobe;Koji Tanaka;Hidehiro Nakamura;Tomonoshin Kanazawa;Takahiro Okawa;Naoto Shiba
    • Clinics in Shoulder and Elbow
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    • v.26 no.2
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    • pp.131-139
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    • 2023
  • Background: Massive rotator cuff tears (RCTs) are complicated by muscle atrophy, fibrosis, and intramuscular fatty degeneration, which are associated with postoperative tendon-to-bone healing failure and poor clinical outcomes. We evaluated muscle and enthesis changes in large tears with or without suprascapular nerve (SN) injury in a rat model. Methods: Sixty-two adult Sprague-Dawley rats were divided into SN injury (+) and SN injury (-) groups (n=31 each), comprising tendon (supraspinatus [SSP]/infraspinatus [ISP]) and nerve resection and tendon resection only cases, respectively. Muscle weight measurement, histological evaluation, and biomechanical testing were performed 4, 8, and 12 weeks postoperatively. Ultrastructural analysis with block face imaging was performed 8 weeks postoperatively. Results: SSP/ISP muscles in the SN injury (+) group appeared atrophic, with increased fatty tissue and decreased muscle weight, compared to those in the control and SN injury (-) groups. Immunoreactivity was only positive in the SN injury (+) group. Myofibril arrangement irregularity and mitochondrial swelling severity, along with number of fatty cells, were higher in the SN injury (+) group than in the SN injury (-) group. The bone-tendon junction enthesis was firm in the SN injury (-) group; this was atrophic and thinner in the SN injury (+) group, with decreased cell density and immature fibrocartilage. Mechanically, the tendon-bone insertion was significantly weaker in the SN injury (+) group than in the control and SN injury (+) groups. Conclusions: In clinical settings, SN injury may cause severe fatty changes and inhibition of postoperative tendon healing in large RCTs. Level of evidence: Level Basic research, controlled laboratory study.

Anti-arthritis Effects of Jeonsaenghwalhyeoltanggamibang through Immune Modulation Modulation (전생활혈탕가미방(全生活血湯加味方)의 면역조절 작용을 통한 항관절염 효능에 관한 연구)

  • Ko, Jeong-Min;Choe, Chang-Min;Cho, Han-Baek;Kim, Song-Baeg
    • The Journal of Korean Obstetrics and Gynecology
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    • v.22 no.2
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    • pp.1-25
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    • 2009
  • Purpose: The purpose of this study is to investigate the anti-arthritis effects of Jeonsaenghwalhyeoltanggamibang(JHTG) on collagen-induced arthritis(CIA) in mice. Methods: To assess the effects of JHTG on CIA in mice, we conducted several experiments such as analysis of arthritis index, cell count of draining lymph node(DLN) and paw joint, measurement of serum antibody levels and observation of the histological changes of joint. Results: 1. JHTG extract had a suppressive effect on the arthritis index of paw joints in CIA mice. 2. JHTG extract increased the total cell number of DLN, and decreased the total cell number of paw joints in CIA mice. 3. JHTG extract increased the absolute number of various cell surface receptors in DLN, and decreased the absolute number of B220+/CD23+ cells in DLN in CIA mice. 4. JHTG extract decreased the absolute number of CD3+, CD4+, CD11b+/Gr-1 cells in paw joint in CIA mice. 5. JHTG extract didn't decrease the absolute number of CD4+/CD25+ cells in paw joints in CIA mice. 6. JHTG extract decreased levels of total IgM in the serum of CIA mice, but had no effect on levels of collagen II specific antibody. 7. JHTG extract decreased the destruction of articular cartilages and collagen fibers and the proliferation of synovial cells in paw joints from CIA mice. Conclusion: These results indicate that JHTG has clinical potential for the treatment of rheumatoid arthritis by modulating the immune response.

The effect of PostGraft$^{TM}$ on implants were installed at the tibia of ovariectomized rats (난소절제술한 백서에 식립한 임플란트에 대한 PostGraft$^{TM}$의 경구 투여 효과)

  • Cho, Chun-Il;Kim, Se-Won;Choi, Yu-Sung;Cho, In-Ho
    • The Journal of Korean Academy of Prosthodontics
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    • v.49 no.4
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    • pp.300-307
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    • 2011
  • Purpose: This study evaluated PostGraft$^{TM}$ which enhances implant stability and bone density. Materials and methods: Forty eight implants were installed at the tibia of ovariectomized rats. The group administrated with PostGraft$^{TM}$ was the experimental group, and the control group was not administrated. Implant stability was evaluated at the $2^{nd}$, $4^{th}$ and $6^{th}$ week by Periotest value, bone mineral density, bone-to-implant contact. These values were analyzed statistically with Mann-Whitney U test (P<.05). Histological analysis was evaluated at the $2^{nd}$, $4^{th}$ and $6^{th}$ week. Results: According to the Periotest$^{(R)}$ measurement, both experimental and control groups showed decrease in values as time elapsed. Greater decrease was observed in the experimental group but there was no significant difference. By examining the radiographic images, both experimental and control groups showed tendency of increase in bone density. Greater increase was seen in the experimental group but there was no significant difference. According to the bone-to-implant contact measurement, both experimental and control groups showed increase in values as time elapsed. Greater increase was seen in the experimental group. At the $2^{nd}$ and $4^{th}$ week, there was no significant difference. But at the $6^{th}$ week, there was significant difference (P<.05). By histological analysis, both experimental and control groups showed increase in bone formation as time elapsed. In addition, greater increase was seen in the experimental group. Conclusion: It could be concluded that the PostGraft$^{TM}$ medicated group showed better results in the bone density and osseointegration.

THE CHANGE OF BONE FORMATION ACCORDING TO MAGNETIC INTENSITY OF MAGNET PLACID INTO TITANIUM IMPLANT SPECIMENS (타이타늄 임플랜트 시편 내부에 설치한 자석의 자성강도에 따른 골형성 변화)

  • Hwang Yun-Tae;Lee Sung-Bok;Choi Dae-Gyun;Choi Boo-Byung
    • The Journal of Korean Academy of Prosthodontics
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    • v.43 no.2
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    • pp.232-247
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    • 2005
  • Purpose. The purposes of this investigation were to discover the possibility of clinical application in the areas of dental implants and bone grafts by investigating the bone formation histologically around specimen which was depending on the intensity of magnetic field of neodymium magnet inside of the specimens. Material and method. 1. Measurement of magnetic intensity - placed the magnet inside of the specimen, and measured the intensity of magnetic field around the 1st thread and 3rd thread of specimen 20 times by using a Gaussmeter(Kanetec Co., Japan). 2. Surgical Procedure - Male rabbit was anesthetised by constant amount of Ketamine (0.25ml/kg) and Rompun (0.25ml/kg). After incising the flat part of tibia, and planted the specimens of titanium implant, control group was stitched without magnet, while experimental groups were placed a magnedisc 500(Aichi Steel Co., Japan) or magnedisc 800(Aichi Steel Co., Japan) into it, fixed by pattern resin and stitched. 3. Management after the surgery - In order to prevent it from the infection of bacteria and for antiinflammation, Gentamycin and Ketopro were injected during 1 week from operation day, and dressed with potadine. 4. Preparation of histomorphometric analysis - At 2, 4 and 8 weeks after the surgery, the animals were sacrificed by excessed Ketamine, and then, specimens were obtained including the operated part and some parts of tibia, and fixed it to 10% of PBS buffer solution. After embedding specimens in Technovit 1200 and B.P solution, made a H-E stain. Samples width was 75$\mu$m . In histological findings through the optical microscope and using Kappa image base program(Olympus Co. Japan), the bone contact ratio and bone area ratio of each parts of specimens were measured and analyzed. 5. Statistical analysis - Statistical analysis was accomplished with Mann Whitney U-test. Results and conclusion. 1. In histomorphometric findings, increased new bone formation was shown in both control & experimental groups through the experiment performed for 2, 4 & 8 weeks. After 4 weeks, more osteoblasts and osteoclasts with significant bone remodeling were shown in experimental groups. 2. In histomorphometric analysis, the bone contact ratios were 38.5% for experimental group 1, 29.5% for experimental group 2 and 11.9% for control group. Experimental groups were higher than control group(p<0.05) (Fig. 6, Table IV). The bone area ratios were 60.9% for experimental group 2, 46.4% for experimental group 1 and 36.0% for control group. There was no significantly statistical difference between experimental groups and control group(p<0.05) (Fig. 8, Table VII) 3. In comparision of the bone contact ratios at each measurement sites according to magnetic intensity, experimental group 2(5.6mT) was higher than control group at the 1st thread (p<0.05) and experimental group 1 (1.8mT) was higher than control group at the 3rd thread(p<0.05) (Fig. 7, Table V, VI). 4. In comparision of the bone area ratios at each measurement sites according to magnetic intensity, experimental group 2(5.6mT) was higher than control group and experimental group 1 (4.0mT) at the 1st thread(p<0.1) and experimental group 2(4.4mT) was higher than experimental group 1 (1.8mT) at the 3rd thread(p<0.1) (Fig. 9, Table IX, X). Experiment group 2 was largest, followed by experiment group l and control group at the 3rd thread of implant. There was a significant difference at the 1st thread of control group & experiment group 2, and at 1st thread & 3rd thread of experiment group 1 & 2, and not at control group experiment group 1.(p<0.1)

Water Extract of Hovenia dulcis Suppressed Lipid Peroxidation and Improved Renal Function in $CCl_4$ Intoxicated Rats ($CCl_4$를 투여한 랫드에서 헛개나무 열수추출물의 지질과산화 억제와 신기능 개선 작용)

  • park Yeun Woo;Yang Si Yang;Lee Min Kyung;Jin Ju Young;Cho Jung Hee;Kim Ki Young
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.18 no.3
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    • pp.868-873
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    • 2004
  • Renal dysfunction could be developed as the secondary disease of liver cirrhosis. Delayed or suppresed lipid peroxidation by the treatment with physiological active substances could be explained as the antioxidative and protective effect in tissue damage. In this study, we investigated an antioxidative effect and renal function improvement of Hovenia dulcis in liver fibrosis(cirrhosis) induced rats. The female Sprague-Dawley rats (180∼210 g) were divided into 3 groups (Normal, AC: CCl₄ mixture treated group, AC-HV: CCl₄ mixture+ Hovenia dulcis treated group) and renal damage was developed by CCl₄ mixture administration in 4 weeks (0.8 ㎖/rat). The tissue of kidney and liver and sera were used for quantitative measurement of enzyme activity, MDA and Hyp. The histological change and gene expression of collagen α1(III) mRNA and a1(IV) mRNA were observed by Masson's trichrome staining and RT-PCR. As a result, the clinical biochemical parameters of liver function (AST and ALT) in sera of AC-HV group showed significantly 46.4% and 104.8% lower (p<0.005), and the level of ALP and BUN as the parameter of protein urine and azotemia showed 17.8 % and 25.8 % lower than in AC group. In AC-HV group, the concentration of MDA in kidney and liver was decreased significantly 15.8% and 21.3% when compared with AC group (p<0.01 -0.005). The content of Hyp in kidney of AC-HV group is merely higher than in AC group, in contrast to liver tissue. The expression of collagen α1(III) mRNA and collagen α1(IV) mRNA was decreased in AC, but both of collagen mRNA in normal and AC-HV group expressed fast similar. More massive lipid droplets, thicker collagen fiber bundles in portal triads and more formation of portal central septum were observed in the liver of AC group than in AC-HV group. In conclusion, CCl₄ mixture intoxication could be developed not only liver fibrosis(cirrhosis) but also renal dysfunction by the massive lipid peroxidation and suppression of interstitial collagen and basement membrane collagen synthesis. And the water extract of Hovenia dulcis may be possessed the antioxidative and protective effect and improvement of kidney function in renal dysfunction induced rats.

Effects of Sujeom Powder Pharmacopuncture Injected at Jung-wan($CV_{12}$) on the Caerulein-induced Acute Pancreatitis in the Rat (중완(中脘)($CV_{12}$) 수념산(手拈散)약침이 Caerulein으로 유발된 흰쥐의 췌장염에 미치는 영향)

  • Kim, In Soo;Jeon, Sang Yun;Jeong, Tae San;Kang, Sung Sun;Jo, Jae Jun;Lee, Young Su
    • Journal of Acupuncture Research
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    • v.29 no.6
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    • pp.35-45
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    • 2012
  • Objectives : This study was designed to investigate Effects of Sujeom powder(SJP) pharmacopuncture Injected at Jung-wan($CV_{12}$) in rats with caerulein-induced acute pancreatitis(AP). Methods : We examined changes of organ weight, histology, immunohistochemistry and gene expression of cycolooxygenase 2(COX-2) in the pancreas. Twenty adult male Sprague-Dawley rats were divided into four groups as follow: normal(Nor), caerulein-induced(Con), caerulein+SJP pharmacopuncture 0.2mL injected at Jung-wan($CV_{12}$)(SA), and caerulein+SJP pharmacopuncture 0.8 mL injected at Jung-wan($CV_{12}$)(SB) groups. Pancreatic tissues of rats from all groups were removed for histological observation and light microscopic examination. Interleukin-6(IL-6) levels were determined spectrophotometrically. Results : The ratio of pancreas/body weights was significantly(p<0.05) increased in the Con, the SA and the SB compared with the Nor, but was slightly decreased in the SA and in the SB groups compared with the Con. Caerulein administration has significantly(p<0.05) increased in the levels of amylase, but the SA, the SB significantly(p<0.05) decreased in the levels of these enzyme. The levels of amylase were increased significantly with caerulein administration, but were inhibited significantly in the SA and in the SB groups. Interleukin-6(IL-6) levels were significantly(p<005) increased in all groups compared with the Nor, especially in the SB. were significantly increased. The levels of Tumor necrosis factor(TNF)-${\alpha}$ levels were significantly increased in all groups compared with the Nor. In the conclusion, the datum of IL-6 and TNF-${\alpha}$ are suggested that the inflamation was still existed actively at a point of measurement(24 hours later). The COX-2 positive materials are observed in the pancreas from the Con, but these positive materials are decreased in the SJP pharmacopuncture at Jung-wan($CV_{12}$) treatment group. Conclusion : SJP pharmacopuncture injected at Jung-wan($CV_{12}$) is potentially capable of limiting pancreatic damage during AP by restoring the fine structure of acinar cells and tissues. Therefore we can say that SJP pharmacopuncture Injected at Jung-wan($CV_{12}$) may have beneficial effects in the treatment of caerulein-induded AP. Further studies about the adequate amount of the SJP pharmacopuncture and about more effective route of administration is still required.

In Vitro Magnetometric Evaluation far Toxicity to Alverolar Macrophage of Arsenic Compounds (In Vitro 자계(磁界) 측정에 의한 비소화합물의 폐포 Macrophage 독성 평가)

  • Cho, Young-Chae
    • Journal of Preventive Medicine and Public Health
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    • v.32 no.4
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    • pp.467-472
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    • 1999
  • Objectives: This study was conducted to evaluate the cytotoxicity of gallium arsenide(GaAs), indium phosphide(InP) and indium arsenide(InAs) all of which are used a$ the semiconductor eletments in semiconductor industry. Methods: Cytotoxicity id the alveolar macrophage was evaluated by the measurement of in vitro magnetometry, LDH release assay and histological examination. Results: The relaxation curves by the in vitro magnetometry showed that GaAs has the cytotoxicity for the alveolar macrophage which is more significant in the higher dosages, while this cytotoxicity is not appeared in the groups added with InP or InAs or PBS. In the decay constant for two minutes after magnetization, GaAs-added groups showed a significant decrease with increasing doses, but both InP- and InAs-added groups did not show any significance. The LDH release assay showed a dose-dependent increasing tendency in the GaAs-, InP- and InAs-added groups. In terms of cellular morphological changes, GaAs-added groups revealed such severe cellular damages as prominent destructions in cell membranes and their morphological changes of nucleus, while InP- and InAs-added groups remained intact in intracellular structures, except for cytoplasmic degenerations. Conclusions: It is suggested that GaAs is more influential to cytotoxicity of alveolar macrophages than InP and InAs.

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Study of Mylabris Phalerata on Anti-cancer Effects in Some Kinds of Cancer Cells (반모가 수종의 인체 암세포에 미치는 영향)

  • Kim, Jin-Sung;Yoon, Sang-Hyub;Ryu, Bong-Ha;Ryu, Ki-Won;Jung, Myung-Chai
    • The Journal of Internal Korean Medicine
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    • v.25 no.2
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    • pp.202-213
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    • 2004
  • Object : Objective: This study was conducted to investigate the anti-cancer effects of Mylabris phalerata (반모) in some kinds of cancer cells. Materials and Methods: Some kinds of cancer cells lines were treated. We used nine kinds of cancer cell lines, such as stomach cancer cells (Kato), lung cancer cells (Calu-1, NCI-H 1395), urinary bladder cancer cells (HS789T), bone cancer cells (Saos-2), brain cancer cells (SK-N-MC), liver cancer cells (Hep-G2), skin cancer cells (Mo-1) and prostate cancer cells (PC-3) with the water decoction of Mylabris phalerata. The histological changes of all cell lines in the media (RPMI-1640) containing the decoction of Mylabris phalerata were observed and we examined cell death assay by trypan blue exclusion testing was examined. Finally, the change of mitochondrial membrane potential was measurd and the inhibitory effect of Mylabris phalerata on cell increase was examined by analyzing the cell cycle. Results: In histologic change all cancer cell lines showed withdrawn and floating appearance that is typical in cellular impairment. Most of the cell lines showed over 50% death rate after 24 hours in trypan blue exclusion tests. Especially the stomach, urinary bladder. brain and liver cell lines showed over 30% death rate after 12 hours. All cell lines treated with Mylabris phalerata were less stained than the control group and the mitochondrial membrane potential in the Mylabris phalerata treated cell lines was markedly lower than that in the control group. The measurement of DNA quantity in all cell lines showed the disappearance of the peak and the thickened left axis, which suggests that all cellular DNA degraded. Conclusion: Mylabris phalerata had cytotoxicity on various kinds of cancer cell lines and the mechanism of that was the impairment of mitochondria by the breakdown of the mitochondrial cell membrane. We propose that this is in part attributable to the destruction of DNA in cancer cells.

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Intragastrically Applicated CCl4-Thiopental Sodium Enhanced Lipid Peroxidation and Liver Fibrosis (Cirrhosis) in Rat: Malonedialdehyde as a Parameter of Lipid Peroxidation Correlated with Hydroxyproline as a Parameter of Collagen Synthesis (Deposition)

  • Kim, Ki-Young;Cho, Syung-Eun;Yu, Byung-Soo
    • Toxicological Research
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    • v.25 no.2
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    • pp.71-78
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    • 2009
  • We investigated the pathogenesis of liver tissue damage during the lipid peroxidation and fibrogenesis with the observation of correlations between the parameters of collagen synthesis (and deposition) and lipid peroxidation in liver fibrosis (cirrhosis) rats. Rats were randomly divided into two groups, normal and $CCl_4$-thiopental sod. intoxicated group. And the one group was treated intragastrically with the mixture of $CCl_4$-thiopental sod. 3 times per week for 3 weeks. The liver tissue and sera were used for the measurement of hydroxyproline (HYP), malonedialdehyde (MDA) and superoxide dismutase (SOD). Biochemical parameters such as aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP), total-bilirubin and blood urea nitrogen (BUN) were measured. Additionally, the expression of collagen ${\alpha}1$(III) and $\beta$-actin mRNA was observed by RTPCR. The histological change in liver tissue was also observed by Masson's trichrome and H&E staining. Correlation analysis was carried by Spearman's rho method. All biochemical parameters except total-bilirubin were significantly higher in the $CCl_4$-thiopental sod. treated group than that of the normal group (p < 0.01). In the $CCl_4$-thiopental sod. treated group, Hyp as a parameter of collagen synthesis (deposition) and MDA as a metabolite of lipid peroxidation, were significantly elevated by 1.98 and 2.11 times higher than that of the normal group (p < 0.001) respectively. The activity of SOD in the $CCl_4$-thiopental sod. treated group is decreased significantly by 44.8% (p < 0.001). And collagen ${\alpha}1$(III) mRNA was more expressed in the $CCl_4$-thiopental sod. treated group than that of the normal group. However, the expression of $\beta$-actin mRNA is showed similar in both of groups. A good correlation was observed between the content of hyp and MDA concentration (r = 0.70, n = 40) in the two groups. And the correlation between the levels of hyp and SOD (r = -0.71, n = 25) is also reliable. However, no correlation were observed between MDA concentration and SOD (r = -0.40, n = 25) in the two groups. Elevated levels of MDA in $CCl_4$-thiopental sod. treated rats indicated enhancement of lipid peroxidation, which is accompanied by a decrease in SOD activity. Moreover, we could confirm that the parameters of collagen synthesis (and deposition) is in good correlation with the metabolite of lipid peroxidation (MDA) and the lipid peroxidation antagonizing enzyme (SOD). Hence, we propose that (1) lipid peroxidation and collagen synthesis (and deposition) could be enhanced by intragastrically application of $CCl_4$-thiopental sod. during a short terms. And (2) the intoxication of $CCl_4$-thiopental sod. could be used for monitoring of lipid peroxidation and collagen synthesis (and deposition) for test of antioxidant and antifibrotic agent.

Compound K improves skin barrier function by increasing SPINK5 expression

  • Park, No-June;Bong, Sim-Kyu;Lee, Sullim;Jung, Yujung;Jegal, Hyun;Kim, Jinchul;Kim, Si-Kwan;Kim, Yong Kee;Kim, Su-Nam
    • Journal of Ginseng Research
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    • v.44 no.6
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    • pp.799-807
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    • 2020
  • Background: The skin acts as a barrier to protect organisms against harmful exogenous agents. Compound K (CK) is an active metabolite of ginsenoside Rb1, Rb2 and Rc, and researchers have focused on its skin protective efficacy. In this study, we hypothesized that increased expression of the serine protease inhibitor Kazal type-5 (SPINK5) may improve skin barrier function. Methods: We screened several ginsenosides to increase SPINK5 gene promoter activity using a transactivation assay and found that CK can increase SPINK5 expression. To investigate the protective effect of CK on the skin barrier, RT-PCR and Western blotting were performed to investigate the expression levels of SPINK5, kallikrein 5 (KLK5), KLK7 and PAR2 in UVB-irradiated HaCaT cells. Measurement of transepidermal water loss (TEWL) and histological changes associated with the skin barrier were performed in a UVB-irradiated mouse model and a 1-chloro-2,4-dinitrobenzene (DNCB)-induced atopic dermatitis-like model. Results: CK treatment increased the expression of SPINK5 and decreased the expression of its downstream genes, such as KLKs and PAR2. In the UVB-irradiated mouse model and the DNCB-induced atopic dermatitis model, CK restored increased TEWL and decreased hydration and epidermal hyperplasia. In addition, CK normalized the reduced SPINK5 expression caused by UVB or DNCB, thereby restoring the expression of the proteins involved in desquamation to a level similar to normal. Conclusions: Our data showed that CK contributes to improving skin-barrier function in UVB-irradiated and DNCB-induced atopic dermatitis-like models through SPINK5. These results suggest that therapeutic attempts with CK might be useful in treating barrier-disrupted diseases.