During involution of the mammary gland after the lactation period, the gland undergoes an extensive epithelial cell death. In our previous study, overexpression of an extracellular proteinase inhibitor (Expi) gene accelerated apoptosis of mammary epithelial cells. Here we found that expression of the cathepsin D gene was induced in the Expi-overexpressed apoptotic cells. To understand the role of cathepsin D in apoptosis, we transfected cathepsin D gene into mammary epithelial HC11 cells and established the stable cell lines overexpressing the cathepsin D gene. We found that overexpression of the cathepsin D gene partially induced apoptosis of mammary epithelial cells. Expression patterns of the cathepsin D gene were examined in mouse mammary gland at various reproductive stages. Expression of the cathepsin D gene was increased during involution stages compared to lactation stages, and highest expression levels were shown at involution on day 4. We also examined expression of the cathepsin D gene in various mouse tissues. Mammary gland at involution on day 2 showed highest levels of cathepsin D mRNA of the mouse tissues that we examined. Liver tissues showed high levels of cathepsin D expression. These results demonstrate that cathepsin D may contribute to the apoptotic process of mammary epithelial cells.
Secretory ducts in the stem of Panax ginseng seedlings are observed with light and electron microscopes to clarify development of the epithelial cells of secretory ducts. Secretory duct initial cell is developed from procambial cell which originated from initial cell is differentiated into ipithelial cell ofsecretory ducts. Intercellular space between the epithelial cells are gradually expanded and differentiated into duct lumen. Disintegrations of epithelial cells occur throughout all the stages of development. The cytoplasm of epithelial cells darken and the epithelial cell wall are lysed, preceding their disintegraton. In the epithelial cell organelles are scattered in the cytoplasm. Development of vcuoles are sparse at the early stage. Starch grains decreased gradually, while lipid droplets increased. Free ribosomes are distributed throughout the cytoplasm and secretory vesicles which originated from rough endoplasmic reticulum and Golgi complex are fused with the plasmalemma. These suggest that the cellular metabolism is active. Microtubules and plasmodesmata are typically observed in the thickened epithelial cell wall. Secretions are accumulated in duct lumen.
Degenerative retinal diseases affect millions of people worldwide, which can lead to the loss of vision. However, therapeutic approaches that can reverse this process are limited. Recent efforts have allowed the possibility of the stem cell-based regeneration of retinal cells and repair of injured retinal tissues. Although the direct differentiation of pluripotent stem cells into terminally differentiated photoreceptor cells comprises one approach, a series of studies revealed the intrinsic regenerative potential of the retina using endogenous retinal stem cells. Muller glial cells, ciliary pigment epithelial cells, and retinal pigment epithelial cells are candidates for such retinal stem cells that can differentiate into multiple types of retinal cells and be integrated into injured or developing retina. In this review, we explore our current understanding of the cellular identity of these candidate retinal stem cells and their therapeutic potential for cell therapy against degenerative retinal diseases. [BMB Reports 2015; 48(4): 193-199]
Objective: Orchastric changes in the mammary glands are vital, especially during lactation. The secretary epithelial cells together with the supporting myoepithelial and stromal cells function cordially to secrete milk. Increase in the number of luminal epithelial cells and a decrease in adipocytes are visible during lactation, whereas the reverse happens in the involution. However, an early involution occurs if the epithelial cells transdifferentiate towards adipocytes during the lactation period. We aimed to inhibit the adipocyte transdifferentiation of luminal cells by restraining the peroxisomal proliferator-activated receptor γ (PPARγ) pathway. Methods: Linolenic acid (LA) and thiazolidinediones (TZDs) induced adipogenesis in mammary epithelial cells were conducted in monolayer, mixed culture as well as in transwell plate co-culture with mammary myoepithelial cells. Results: Co-culture with myoepithelial cells showed higher adipogenic gene expression in epithelial cells under LA+TZDs treatment. Increase in the expressions of PPARγ, CCAAT/enhancer-binding protein α and vimentin in both mRNA as well as protein levels were observed. Whereas, bisphenol A diglycidyl ether treatment blocked LA+TZDs induced adipogenesis, as it could not show a significant rise in adipose related markers. Although comparative results were found in both mixed culture and monolayer conditions, co-culture technic was found to work better than the others. Conclusion: Antagonizing PPARγ pathway in the presence of myoepithelial cells can significantly reduce the adipogenisis in epithelial cells, suggesting therapeutic inhibition of PPARγ can be considered to counter early involution or excessive adipogenesis in mammary epithelium in animals.
Lactobacillus species in the female genital tract are thought to act as a barrier to infection. Several studies have demonstrated that lactobacilli can adhere to vaginal epithelial cells. However, little is known about how the adherence of lactobacilli to vaginal epithelial cells affects the acidity, cell viability, or proliferation of the lactobacilli themselves or those of vaginal epithelial cells. Lactobacillus acidophilus was co-cultured with immortalized human vaginal epithelial cells (MS74 cell line), and the growth of L. acidophilus and the acidity of the culture medium were measured. MS74 cell density and viability were also assessed by counting cell numbers and observing the cell attachment state. L. acidophilus showed exponential growth for the first 6 hr until 9 hr, and the pH was maintained close to 4.0-5.0 at 24 hr after culture, consistent with previous studies. The growth curve of L. acidophilus or the pH values were relatively unaffected by co-culture with MS74 cells, confirming that L. acidophilus maintains a low pH in the presence of MS74 cells. This co-culture model could therefore potentially be used to mimic vaginal conditions for future in vitro studies. On the other hand, MS74 cells co-cultured with L. acidophilus more firmly attached to the culture plate, and a higher number of cells were present compared to cells cultured in the absence of L. acidophilus. These results indicate that L. acidophilus increases MS74 cell proliferation and viability, suggesting that lactobacilli may contribute to the healthy environment for vaginal epithelial cells.
Foreign molecules, including viruses and bacteria-derived toxins, can also induce airway inflammation. However, to the best of our knowledge, the roles of these molecules in the development of airway inflammation have not been fully elucidated. Herein, we investigated the precise role and synergistic effect of virus-mimicking double-stranded RNA (dsRNA) and staphylococcal enterotoxin B (SEB) in macrophages and epithelial cells. To identify cytokine expression profiles, both the THP-1-derived macrophages and BEAS-2B epithelial cells were stimulated with dsRNA or SEB. A total of 21 cytokines were evaluated in the culture supernatants. We observed that stimulation with dsRNA induced cytokine production in both cell types. However, cytokine production was not induced in SEB-stimulated epithelial cells, compared to the macrophages. The synergistic effect of dsRNA and SEB was evaluated observing cytokine level and intracellular phospho-signaling. Fifteen different types were detected in high-dose dsRNA-stimulated epithelial cells, and 12 distinct types were detected in macrophages; those found in macrophages lacked interferon production compared to the epithelial cells. Notably, a synergistic effect of cytokine induction by co-stimulation of dsRNA and SEB was observed mainly in epithelial cells, via activation of most intracellular phosphor-signaling. However, macrophages only showed an accumulative effect. This study showed that the type and severity of cytokine productions from the epithelium or macrophages could be affected by different intensities and a combination of dsRNA and SEB. Further studies with this approach may improve our understanding of the development and exacerbation of airway inflammation and asthma.
Histological and histochemical studies were made on the lining epithelia of the various ducts in epidymis of the Rooster and absorptive function of the canal epithelial cells in the Rooster epididymis were also investigated after administration of India ink. The results obtained were summerized as follows; 1. Epihtelium lining the rate testis was mainaly composed of single later of cuboidal cells, and was partially composed of flattened squamous or low columnar cells. Efferential ductules were characterized by having many villous projections orrfolds which extened into the lumen, and were lined by stereociliated pseudostratified epithelium which consisted of manily ciliated columnar cells, a few scattered clear cells and basal cells. Connecting ductules were lined by ciliated pseudostriatified colummnar epithelium in which ciliated columnar cells, clear cells and basal cells were noted. Epididymal ducts were lined by pseudostratified epidhelium in which columnar and basal cells were noted. 2. PAS-granules, saliva resistant were noted mainly in the epithelial cells of efferential and connecting ductules. 3. Sudan black B stained heavily the granules in the epithelial cells of sufferential and connecting ductules. 4. The granules reactive to acid phosphatase most abundant in the epithelial cells of efferential ductules and were lesser amount in the epithelial cells of connecting ductules where as very few or no granules were seen in the rest of the ducts. 5. Alkaline phosphatase activity was most prominent but discontinuous in the luminal surface of the epithelium of efferential ductules and less marked in the connecting ductless. No enzyme activity was noted in the canal epithelium of epididyml duct. 6. India ink granules were most numerous in the epithlial cells of efferential ductules and were a few in connecting ductules. Very few or no granules of India ink were noted in the other types of the ducts. India ink granules in the epithelium increased gradually as the time after the administration of India ink (one up to twenty-nine hours) has proceeded. From those results it is suggested that epithelial cells of efferential and connecting ductules have active absorptive function, whereas the rest of duct system in the epididymis of the Rooster may be the mere pathway of the seminal fluid without significant modification of its constituents.
The trans-differentiation potential of mesenchymal stem cells (MSCs) is employed, but there is little understanding of the cell source-dependent trans-differentiation potential of MSCs into corneal epithelial cells. In the present study, we induced trans-differentiation of MSCs derived from umbilical cord matrix (UCM-MSCs) and from dental tissue (D-MSCs), and we comparatively evaluated the in vitro trans-differentiation properties of both MSCs into corneal epithelial-like cells. Specific cell surface markers of MSC (CD44, CD73, CD90, and CD105) were detected in both UCM-MSCs and D-MSCs, but MHCII and CD119 were significantly lower (P < 0.05) in UCM-MSCs than in D-MSCs. In UCM-MSCs, not only expression levels of Oct3/4 and Nanog but also proliferation ability were significantly higher (P < 0.05) than in D-MSCs. In vitro differentiation abilities into adipocytes and osteocytes were confirmed for both MSCs. UCM-MSCs and D-MSCs were successfully trans-differentiated into corneal epithelial cells, and expression of lineage-specific markers (Cytokeratin-3, -8, and -12) were confirmed in both MSCs using immunofluorescence staining and qRT-PCR analysis. In particular, the differentiation capacity of UCM-MSCs into corneal epithelial cells was significantly higher (P < 0.05) than that of D-MSCs. In conclusion, UCM-MSCs have higher differentiation potential into corneal epithelial-like cells and have lower expression of CD119 and MHC class II than D-MSCs, which makes them a better source for the treatment of corneal opacity.
Platelet activating factor (PAP) has been known as implicating as one of potent inflammatory mediators and reported 0 be involved in inflammation and allergy. PAF induces ciliary dysfunction and epithelial damage of human paranasal sinus mucosa in vitro. However, several recent papers have reported that PAF may not readily damage the airway epithelium. The aim of this study was to investigate the ultrastructural evidence to elucidate the pathogenesis of epithelial damage induced by PAF. Sixteen $\mu\textrm{g}$ g of PAF was applied into the maxillary sinuses of 6 rabbits. Rabbits were divided into 2 subgroups along with time interval at 1st and 3rd experimental day, and sinus mucosae were taken for the histopathologic study using electron microscopy. At 1st day, epithelial cells showed no ultrastructural change. Ultrastructures of the cilia were well preserved. Subepithelial space showed no evidence of the infiltration of inflammatory cells. Intravascular platelet aggregation and swelling of endothelial cells were evident. At 3rd day, epithelial cells showed vacuolar degeneration. Fusion of cilia forming giant cilia and focal loss of cilia were evident. Eosinophils were infiltrated in subepithelial and intraepithelial space. Swelling of endothelial cells, and migration of inflammatory cells into the connective tissue were evident. This study implies that epithelial damage induced by PAF may be secondary to the cytotoxicity of mobilized eosinophils rather than direct cytotoxicity of PAF.
Objectives : Intestinal epithelial cell damage is closely associated with various intestinal diseases, such as Inflammatory Bowel Disease (IBD), Celiac Disease and Gastroenteritis, and it plays a crucial role in the development and progression of intestinal diseases. Therefore, it is important to develop drugs that target protection of intestinal epithelial cells. Here, we aimed to investigated whether Bambusae Caulis in Liquamen (BCL) against t-BHP induced oxidative stress injury in human intestinal epithelial cells and to explore the underlying molecular mechanism. Methods : In this study, we performed MTT assay, measurement of ROS generation, and immunoblot analysis to determine the cytoprotective efficacy in HT29 cells (human colorectal adenocarinoma cell line with epithelial morphogy). Results : First, we checked that BCL was not cytotoxic up to concentration 30 ㎍/mL in HT29 cells. Then, we confirmed that BCL inhibited t-BHP-induced ROS and cell death. BCL also reversed the expression of proteins associated apoptosis. Next, to confirm the relationship between efficacy of BCL and Nrf2, we conducted experiments using siNrf2. Asresult, the effects of inhibiting ROS production and cell death of BCL was reversed by siNrf2. Conclusion : BCL prevents t-BHP-induced oxidative stress and apoptosis. And the efficacy of BCL is related to Nrf2 activation.
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