• Title/Summary/Keyword: $\alpha$-SMA

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Effects of Electrical Stimulation on Wound Healing and Skin Mast Cells in Streptozotocin-Induced Diabetic Rats (스트렙토조토신 유도 당뇨 흰쥐에서 전기자극이 상처치유와 피부 비만세포에 미치는 영향)

  • Jekal, Seung-Joo;Lee, Kyung-Sun;Chung, Ok-Bong;Lee, Jae-Hyoung
    • Korean Journal of Clinical Laboratory Science
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    • v.40 no.2
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    • pp.118-128
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    • 2008
  • The aim of this study was to investigate the effect of electrical stimulation on healing of impaired wound and alteration of mast cells in experimental diabetic rats. Thirty male Sprague-Dawley rats were divided into three groups : incision (control), diabetes+incision (diabetes) and diabetes + incision + electrical stimulation (D/ES). Diabetes was induced in rats by streptozotocin (STZ) injection (60 mg/kg, one time) and 20 mm length incision wounds were created on the back after shaving hair. The electrical stimulation rats were treated with a current intensity of 30~50 V at 120 pps and $140{\mu}s$ for 10 days from 3 days after STZ injection. The lesion and adjacent skin tissues were fixed with 10% buffered formalin, embedded with paraffin. For wound healing analysis, hematoxylin-eosin (HE) and picrosirius red staining were performed. Mast cells (MC) were stained with toluidine blue (pH 0.5) and quantified at ${\times}200$ using a light microscope. The density of keratinocyte proliferation and microvessels in skin tissues were analyzed using a computerized image analysis system on sections immunostained with proliferative cell nuclear antigen (PCNA) and ${\alpha}$-smooth muscle actin (${\alpha}$-SMA), respectively. The results showed that the wound healing rate, collagen density and neoepidermis thickness, density of PCNA-positive cells and density of ${\alpha}$-SMA-positive vessels were significantly higher in D/ES rats than in diabetic rats. The density of MCs and degranulated MCs in D/ES rats were also significantly higher than those in diabetic rats. These findings suggest that the electrical stimulation may promote the tissue repair process by accelerating collagen production, keratinocyte proliferation and angiogenesis in the diabetic rats, and MCs are required for wound healing of skin in rats.

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Quantitative Changes in Tumor-Associated M2 Macrophages Characterize Cholangiocarcinoma and their Association with Metastasis

  • Thanee, Malinee;Loilome, Watcharin;Techasen, Anchalee;Namwat, Nisana;Boonmars, Thidarut;Pairojkul, Chawalit;Yongvanit, Puangrat
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.7
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    • pp.3043-3050
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    • 2015
  • The tumor microenvironment (TME) includes numerous non-neoplastic cells such as leukocytes and fibroblasts that surround the neoplasm and influence its growth. Tumor-associated macrophages (TAMs) and cancerassociated fibroblasts (CAFs) are documented as key players in facilitating cancer appearance and progression. Alteration of the macrophage (CD68, CD163) and fibroblast (${\alpha}-SMA$, FSP-1) cells in Opisthorchis viverrini (Ov) -induced cholangiocarcinoma (CCA) was here assessed using liver tissues from an established hamster model and from 43 human cases using immunohistochemistry. We further investigated whether M2-activated TAMs influence CCA cell migration ability by wound healing assay and Western blot analysis. Macrophages and fibroblasts change their phenotypes to M2-TAMs (CD68+, CD163+) and CAFs (${\alpha}-SMA+$, FSP-1+), respectively in the early stages of carcinogenesis. Interestingly, a high density of the M2-TAMs CCA in patients is significantly associated with the presence of extrahepatic metastases (p=0.021). Similarly, CD163+ CCA cells are correlated with metastases (p=0.002), and they may be representative of an epithelial-to-mesenchymal transition (EMT) with increased metastatic activity. We further showed that M2-TAM conditioned medium can induce CCA cell migration as well as increase N-cadherin expression (mesenchymal marker). The present work revealed that significant TME changes occur at an early stage of Ov-induced carcinogenesis and that M2-TAMs are key factors contributing to CCA metastasis, possibly via EMT processes.

Effect of Atractylodis Rhizoma Alba water extract on streptozotocin-induced diabetes in rats (백출이 streptozotocin 유발 당뇨흰쥐에서 췌장 및 신장에 미치는 영향)

  • Han, Yun-Kyung;Park, Yong-Ki
    • The Korea Journal of Herbology
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    • v.26 no.4
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    • pp.23-30
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    • 2011
  • Objectives : This study aimed to evaluate the effect of Atractylodis Rhizoma Alba water extract on streptozotocin (STZ)-induced diabetes in rats. Methods : Male Sprague-Dawley rats were divided into four groups ; normal, STZ-control and Atractylodis Rhizoma Alba (A) water extract-administrated group. Rats in which diabetic was induced by intraperitonal injection with STZ(60 mg/kg body weight). STZ-induced diabetic rats were orally administrated A extract daily for 5 weeks at doses of 200 or 500 mg/kg. Fasting blood glucose, total cholesterol, triglyceride and blood urea nitrogen were measured in sera of rats. Total volume of urine and urinary creatinine were also measured. Histopathological examination and immunohistochemical staining for the expression of insulin and ${\alpha}$-SMA in pancreas and kidney were performed, respectively. Results : There were no differences in body and kidney weights between STZ-control and A extract-administrated groups. However, serum triglyceride level was significantly decreased in A extract-administrated groups compared with those of STZ-control group. Histopathological analysis of pancreas and kidney revealed increased the number of islets and insulin-positive beta-cells in pancreas, and decreased morphological changes of glomerulus and ${\alpha}$-SMA expression in kidney after the administration of A extract. Conclusions : These results suggest that Atractylodis Rhizoma Alba has a biological action on STZ-induced diabetes in rats via decreasing the serum levels of total triglyceride, and suppressing the morphological changes of pancreas and kidney.

Local anesthesia of the temporomandibular joint to reduce pain during mouth opening for dental treatment in a patient with spinal muscular atrophy

  • Chi, Seong In;Kim, Hyun Jeong;Seo, Kwang-Suk;Lee, Jong Ho;Chang, Juhea
    • Journal of Dental Anesthesia and Pain Medicine
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    • v.16 no.2
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    • pp.137-140
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    • 2016
  • Spinal muscular atrophy (SMA) is an autosomal recessive, severe neuromuscular disorder in which degeneration of alpha motor neurons in the spine progressively weakens and ultimately paralyzes the proximal muscles. It occurs in one per 6,000-10,000 infants, and is a genetic disorder with the second-highest mortality rate worldwide. An 18-year-old male patient with SMA was referred for general anesthesia for difficulty in performing dental treatment due to limited mouth opening caused by temporomandibular joint (TMJ) pain. However, the patient had a high risk of general anesthesia complications, so TMJ pain during mouth opening was reduced through local anesthesia of the TMJ. Fortunately, the anesthesia was successful in reducing pain during mouth opening, enabling the patient to receive dental treatment with an adequate mouth opening.

Anti-Fibrotic Effects of DL-Glyceraldehyde in Hepatic Stellate Cells via Activation of ERK-JNK-Caspase-3 Signaling Axis

  • Md. Samsuzzaman;Sun Yeou Kim
    • Biomolecules & Therapeutics
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    • v.31 no.4
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    • pp.425-433
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    • 2023
  • During liver injury, hepatic stellate cells can differentiate into myofibroblast-like structures, which are more susceptible to proliferation, migration, and extracellular matrix generation, leading to liver fibrosis. Anaerobic glycolysis is associated with activated stellate cells and glyceraldehyde (GA) is an inhibitor of glucose metabolism. Therefore, this study aimed to investigate the anti-fibrotic effects of GA in human stellate LX-2 cells. In this study, we used cell viability, morphological analysis, fluorescence-activated cell sorting (FACS), western blotting, and qRT-PCR techniques to elucidate the molecular mechanism underlying the anti-fibrotic effects of GA in LX-2 cells. The results showed that GA significantly reduced cell density and inhibited cell proliferation and lactate levels in LX-2 cells but not in Hep-G2 cells. We found that GA prominently increased the activation of caspase-3/9 for apoptosis induction, and a pan-caspase inhibitor, Z-VAD-fmk, attenuated the cell death and apoptosis effects of GA, suggesting caspase-dependent cell death. Moreover, GA strongly elevated reactive oxygen species (ROS) production and notably increased the phosphorylation of ERK and JNK. Interestingly, it dramatically reduced α-SMA and collagen type I protein and mRNA expression levels in LX-2 cells. Thus, inhibition of ERK and JNK activation significantly rescued GA-induced cell growth suppression and apoptosis in LX-2 cells. Collectively, the current study provides important information demonstrating the anti-fibrotic effects of GA, a glycolytic metabolite, and demonstrates the therapeutic potency of metabolic factors in liver fibrosis.

Identification of a Novel SNP Associated with Meat Quality in C/EBP${\alpha}$ Gene of Korean Cattle

  • Shin, S.C.;Kang, M.J.;Chung, E.R.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.4
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    • pp.466-470
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    • 2007
  • CCAAT/enhancer binding protein ${\alpha}$($C/EBP{\alpha}$) plays an important role in lipid deposition and adipocyte differentiation. In order to find genetic markers to improve the meat quality of Korean cattle, the bovine $C/EBP{\alpha}$ gene was chosen as a candidate gene to investigate its association with carcass and meat quality traits in Korean cattle. A single nucleotide polymorphism (SNP) was identified at position 271 (A/C substitution) of coding region in the $C/EBP{\alpha}$ gene. A PCR-RFLP procedure with restriction enzyme SmaI was developed for determining the marker genotypes. The frequencies of alleles C and A and were 0.374 and 0.626, respectively. The genotype frequencies for CC, AC and AA were 12.9, 49.0 and 38.1%, respectively, in Korean cattle population. The frequencies of genotype were in agreement with Hardy-Weinberg equilibrium. Association analysis indicated that the gene-specific SNP marker of $C/EBP{\alpha}$ showed a significant association with marbling score (p<0.05). The animals with AA genotype had higher marbling score than those with the AC or CC genotype. Although further studies are needed to validate our results, the $C/EBP{\alpha}$ gene could be useful as a genetic marker for carcass and meat quality traits in Korean cattle.

Identification of a Technique Optimized for the Isolation of Spermatogonial Stem Cells from Mouse Testes

  • Han, Na Rae;Park, Hye Jin;Lee, Hyun;Yun, Jung Im;Choi, Kimyung;Lee, Eunsong;Lee, Seung Tae
    • Journal of Embryo Transfer
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    • v.33 no.4
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    • pp.327-336
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    • 2018
  • To date, there are no protocols optimized to the effective separation of spermatogonial stem cells (SSCs) from testicular cells derived from mouse testes, thus hindering studies based on mouse SSCs. In this study, we aimed to determine the most efficient purification method for the isolation of SSCs from mouse testes among previously described techniques. Isolation of SSCs from testicular cells derived from mouse testes was conducted using four different techniques: differential plating (DP), magnetic-activated cell sorting (MACS) post-DP, MACS, and positive and negative selection double MACS. DP was performed for 1, 2, 4, 8, or 16 h, and MACS was performed using EpCAM ($MACS^{EpCAM}$), Thy1 ($MACS^{Thy1}$), or GFR ${\alpha}1$ ($MACS^{GFR{\alpha}1}$) antibodies. The purification efficiency of each method was analyzed by measuring the percentage of cells that stained positively for alkaline phosphatase. DP for 8 h, $MACS^{Thy1}$ post-DP for 8 h, $MACS^{GFR{\alpha}1}$, positive selection double $MACS^{GFR{\alpha}1/EpCAM}$, and negative selection double $MACS^{GFR{\alpha}1/{\alpha}-SMA}$ were identified as the optimal protocols for isolation of SSCs from mouse testicular cells. Comparison of the purification efficiencies of the optimized isolation protocols showed that, numerically, the highest purification efficiency was obtained using $MACS^{GFR{\alpha}1}$. Overall, our results indicate that $MACS^{GFR{\alpha}1}$ is an appropriate purification technique for the isolation of SSCs from mouse testicular cells.

Construction of an Escherichia-Pseudomonas Shuttle Vector Containing an Aminoglycoside Phosphotransferase Gene and a lacZ' Gene for $\alpha$-Complementation

  • Lee, Bheong-Uk;Hong, Ja-Heon;Kahng, Hyung-Yeel;Oh, Kye-Heon
    • Journal of Microbiology
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    • v.44 no.6
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    • pp.671-673
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    • 2006
  • A new 4.87 kb Escherichia-Pseudomonas shuttle vector has been constructed by inserting a 1.27 kb DNA fragment with a replication origin of a Pseudomonas plasmid pRO1614 into the 3.6 kb E. coli plasmid pBGS18. This vector, designated pJH1, contains an aminogly-coside phosphotransferase gene (aph) from Tn903, a lacZ' gene for $\alpha$-complementation and a versatile multiple cloning site possessing unique restriction sites for EcoRI, SacI, KpnI, SmaI, BamHI, XbaI, SalI, BspMI, PstI, SphI, and HindIII. When pJH1 was transformed into E. coli DHS${\alpha}$ and into P. putida HK-6, it was episomally and stably maintained in both strains. In addition, the enhanced green fluorescent protein (EGFP) gene which was transcriptionally cloned into pJH1 rendered E. coli cells fluorescence when its transformants were illuminated at 488 nm.

Tungtungmadic Acid Isolated from Salicornia herbacea Suppresses the Progress of Carbon Tetrachloride-induced Hepatic Fibrosis in Mice

  • Chung, Young-Chul;Choi, Jae-Ho;Oh, Kyo-Nyeo;Chun, Hyo-Kon;Jeong, Hye-Gwang
    • Toxicological Research
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    • v.22 no.3
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    • pp.267-273
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    • 2006
  • Tungtungmadic acid(3-caffeoyl, 4-dihydrocaffeoyl quinic acid: CDCQ) is a new chlorogenic acid derivative isolated from the Salicornia herbacea. The suppressive effects of CDCQ on the progress of acute carbon tetrachloride($CCl_4$)-induced hepatic fibrosis were investigated in mice. CDCQ significantly suppressed $CCl_4$-induced hepatic necrosis and inflammation, as determined by serum enzymatic activities of alanine and aspartate aminotransferase and serum TNF-$\alpha$ levels in a dose-dependent manner. In addition, increased hepatic lipid peroxidation and fibrosis after acute $CCl_4$ treatment were suppressed by the administration of CDCQ. CDCQ also significantly prevented the elevation of hepatic hydroxyproline and collagen content and ${\alpha}$-smooth muscle actin(${\alpha}$-SMA) expression in the liver of $CCl_4$-intoxicated mice. These results suggest that the suppressive effects of CDCQ against the acute $CCl_4$-induced hepatic fibrosis possibly related to its ability to block both hepatic inflammation and the activation of hepatic stellate cells.

Influence of Heat Treatment on Transformation Characteristics and Shape Recovery in Fe-X%/Mn-5Cr-5Co-4Si Alloy Ribbons (Fe-X%Mn-5Cr-5Co-4Si 합금 리본의 변태특성 및 형상기억능에 미치는 열처리 영향)

  • Kang, H.W.;Jee, K.K.;Jang, W.Y.;Kang, J.W.
    • Journal of the Korean Society for Heat Treatment
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    • v.14 no.3
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    • pp.160-166
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    • 2001
  • The change of ribbon geometry, microstructure and shape recovery with Mn contents, wheel speed and various annealing temperature have been studied in Fe-X%Mn-5Cr-5Co-4Si (X%=15, 20, 24) shape memory alloy (SMA) ribbons rapidly solidfied by single roll chill-block melt-spinning process. The thickness and width of melt-spun ribbons are reduced, results in refining and uniformalizing grains with increasing wheel speed. In the ribbons melt-spun at a wheel speed of 15m/sec, both ${\varepsilon}$ and ${\alpha}^{\prime}$martensites are formed in ribbon 1 (15.5wt%Mn), while only ${\varepsilon}$ martensite is revealed in ribbon 2 (20.2wt%Mn) and ribbon 3 (23.5wt%Mn). The volume fraction of ${\varepsilon}$ martensite is decreased with increasing Mn contents, and those of ${\varepsilon}$ as well ${\alpha}^{\prime}$martensites are increased due to thermal stress relief and grain growth with increasing annealing temperature. Ms temperatures of the ribbons 1, 2 and 3 are fallen with increasing Mn contents. $M_s$ temperatures of the ribbons 1, 2 and 3 annealed at $300^{\circ}C$ for 3 min are risen abruptly, but are nearly constant even at higher annealing temperature, i.e., 400, 500 and $600^{\circ}C$ for 3 min. Shape recovery of the ribbons 1, 2 and 3 increased 30%, 52% and 69% with Mn contents, respectively. Shape recovery of ribbon 1 (15.5wt%Mn) formed ${\varepsilon}$ and ${\alpha}^{\prime}$martensites decreased because of the presence of ${\alpha}^{\prime}$martensite but those of ribbon 2 (20.2wt%Mn) and ribbon 3 (23.5wt%Mn) formed ${\varepsilon}$ martensite increased with increasing annealing temperature.

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