Purpose : This study was performed to investigate the effects of Sobokchukeo- Tang(SCT) on the experimentally-induced endometriosis in rats. Materials and Methods : Endometriosis was induced via the surgical autotransplantation technique in rats. A laparotomy was performed and a $4\;{\times}\;4\;mm$ of the right uterine horn was resected incised and sutured to the peritoneum. And the animals divided into control(n=8) and SCT-treated group(n=8). SCT(1,000 mg/head) was administered orally for 15 days after operation. The weights(body, left uterus, and ovaries) and concentrations of cytokines(MCP-1,$TNF-{\alpha}$, $IL-l{\beta}$) were measured. Histopathology, immunohistochemistry for COX-2, and histochemistry for mast cells of the transplanted uterine tissues were performed. Results : - The $volume(mm^3)$ of transplanted uterine tissues of SCT-treated group$(92.88{\pm}41.89)$ was significantly(p<<0.01) decreased than the control group $(404.50{\pm}317.68)$. The concentration(pg/ml) of MCP-1 in ascites of SCT-treated group$(5,256{\pm}1,209)$ was significantly(p<<0.001) decreased than the control group$(8,632{\pm}1,245)$. - The concentration(pg/ml) of $TNF-{\alpha}$ in ascites of SCT-treated group$(521.8{\pm}306.1)$ was significantly(p<<0.01) decreased than the control group$(1,245.2{\pm}362.2)$. - The percentage of COX-2 positive epithelial layer in transplanted uterine tissues of SCT-treated group$(25.0{\pm}7.3)$ was significantly(p<<0.001) decreased than the control group$(50.2{\pm}8.2)$. - The number of mast cells in the stroma of transplanted uterine tissues of SCT-treated group$(16.5{\pm}6.8)$ was significantly(p<<0.05) decreased than the control group$(26.0{\pm}7.7)$. - The number of mast cells in the periphery of transplanted uterine tissues of control group$(71.3{\pm}18.5)$ was significantly(p<<0.01) decreased than the control group$(109.3{\pm}30.2)$. - Proliferation of epithelia, infiltration of inflammatory cells, and microanglogenesis in transplanted uterine tissues of treated group were weakly observed than the control group. Conclusion : From the above results, Sobokchukeo-Tang(SCT) has an inhibitory effect on the development of transplanted uterine tissue in rats and it is related to the decreased concentration of MCP-1 and $TNF-{\alpha}$, and decreased expression of COX-2, and decreased infiltration of mast cells by administration of Sobokchukeo-Tang.
Kim, Chul-Soo;Lee, Dong-Nyung;Seo, Il-Bock;Kim, Hyung-Jun
The Journal of Korean Obstetrics and Gynecology
/
v.20
no.1
/
pp.61-83
/
2007
Purpose: This study was performed to investigate the effects of Keukhachukeo-Tang (KCT) on the development of experimentally-induced endometriosis in rats. Methods : Endometriosis was induced in rats by autotransplanting uterine tissue to the peritoneum and devided them into three groups: (1) sham-operated group (n=8), (2) surgically induced endometriosis and untreated control group (n =8), (3) surgically induced endometriosis and KCT treated group. KCT (1,200 mg/head) was orally administrated for 15 days after operation. Then we measured the body weight, the volumes of endometriotic implants. The weight (body, left uterus and ovaries) and concentrations of cytokines (MCP-1, TNF-${\alpha}$, IL-l${\beta}$) in serum and peritoneal fluid were also measured. Histopathology, immunohistochemistry for COX-2, and histochemistry for mast cells in transplanted uterine tissue were performed. Results : - The volumes(mm$^3$) of endometriotic implants in KCT-treated group (107${\pm}$66) were significantly decreased (p<0.05) compared with control group (405${\pm}$318). - The contents(pg/ml) of MCP-1 in peritoneal fluid in KCT-treated group (6,940${\pm}$893) were significantly decreased (p<0.01) compared with control group (8,632${\pm}$1,245). - The contents(pg/ml) of TNF-${\alpha}$ in peritoneal fluid in KCT-treated group (847${\pm}$330) were significantly decreased (p<0.05) compared with control group (1,245${\pm}$362). - The percentages(%) of positive epithelial layers for COX-2 in KCT-treated group (31${\pm}$10) were significantly decreased (p<0.01) compared with control group (50${\pm}$8). - The numbers of mast cells in adjacent tissue of transplanted uterine tissue in KCT-treated group (69${\pm}$18) were significantly decreased (p<0.01) compared with control group (109${\pm}$30). - The numbers of mast cells in stroma of transplanted uterine tissue in KCT-treated group(16${\pm}$5) were significantly decreased (p<0.01) compared with control group (26${\pm}$8). - Histopathologically, proliferation of endometriotic epithelia and stroma, and infiltration of inflammatory cells in transplanted uterine tissue of KCT-treated group were weakly observed than those of control group. Conclusion : From the above results, Keukhachukeo-Tang (KCT) have inhibiting effects on the development of transplanted uterine tissue. And these effects are related to the decreased concentration of MCP-1 and TNF-${\alpha}$, decreased expression of COX-2, and decreased infiltration of mast cells by administration of Keukhachukeo-Tang.
The development of trachea in fetuses on 60, 90 and 120 days of gestation and neonates of Korean native goats was investigated by microscopy and scanning and transmission electron microscopy. The results were summarized as follows; Light microscopic studies: 1. In the 60-day-old fetuses, the tracheal walls were differentiated and divided into four layers of the mucosa, submucosa, muscle and cartilage, and adventitia. The tracheal epithelium is composed of stratified ciliated columnar epithelium at 60- and 90-day-old fetuses while the epithelium observed at 120-day-old fetuses was pseudostraified ciliated colummar epithelium. 2. In the 90-day-old fetuses, tracheal glands extended into the submucosa and peripheral area of the tracheal cartilage. The blood vessels were observed in the submucosa and adventitia. The elastic and collagenous fibers were observed in the tracheal walls. 3. In the neonates, the tracheal walls consisted of mucosa with well-developed folds, submucosa, tracheal glands, muscle and cartilage, collagenous and elastic fibers, and adventitia, which were more developed than those of 120-day-old fetuses. The tracheal epithelium was developed as that in adults. Scanning electron microscopic studies: 4. In the 60-day-old fetuses, most of tracheal epithelial cells were nonciliated but short microvilli were sporadically observed on the luminal surface. On rare occasions, a few cells have solitary cilium. 5. In the 90-day-old fetuses, the ciliated cells appeared increasingly and cilia elongated longer than those of 60-day-old fetuses. 6. In the 120-day-old fetuses, the nonciliated cells covered with microvilli in dome-shape were barriered by thick carpet of cilia. The nonciliated cells also have many papillary projectons on the apical surface. 7. In the neonates, the nonciliated cells in tracheal epithelium were covered compactly with numerous cilia, and many secretory droplets were found on the cilia. Transmission electron microscopic studies: 8. In the 60-day-old fetuses, nonciliated cells of the tracheal epithelium contain large amounts of glycogen granules in the supernuclear and subnuclear areas meanwhile a few cell organelles were formed. Cilia were well formed along the apical cell membranes of the ciliated cells. Also found in the ciliated cells were basal corpuscles, mitochondria and short chains in granular endoplasmic reticulum(GER). Between the epithelial cells presented were well-defined junctional complex with zonula occludens and desmosomes. The nuclei were variable in size and shape. The more developed nucleoli were observed conspicuosly. 9. In the 90-day-old fetuses, nonciliated cells contained large glycogen granules. Accumulated glycogen granules were observed in the subnuclear and supranuclear portion of the cytoplasm. A few short microvilli were covered with glycocalyx. Ciliated cells contained numerous mitochondria and short chains of GER. 10. In the 120-day-old fetuses, the ciliated cells contained numerous mitochondria, abundant short chains of GER and nucleoli. Nonciliated cells contained some Golgi complex and mitochondria. The cell borders were well-defined and distinct junctional complex with zonula occludens, desmosomes, and interdigitorum. 11. In the neonates, well-developed goblet cells were observed in the tracheal epithelium. Ultrastructures of ciliated and nonciliated cells on the tracheal epithelia were similar in pattern as those in adults.
The objective of the present study was to investigate the uptake process of 4-Phenylazobenzoxycarbonyl-Pro-Leu-Gly-Pro-D-Arg (Pz-peptide), a hydrophilic and collagenase-labile pentapeptide, by isolated hepatocytes. For comparison, the uptake of Pz-peptide by Caco-2 cells and colonic cells, two known paracellular routes of Pz-peptide, was also evaluated. A simple and sensitive reversed-phase HPLC assay method using UV detection has been developed. The coefficient of variation for all the criteria of validation were less than 15%. The method was, therefore, considered to be sutable for measuring the concentration of Pz-peptide in the biological cells. Pz-peptide was extensively uptaked into hepatocytes. The initial velocity of Pz-peptide uptake assessed from the initial slope of the curve was plotted as Eadie-Hofstee plots. The maximum velocity ($V_{max}$) and the Michaelis constant ($K_m$) were 0.190$\pm$0.020 $nmol/min/10^6$ cells and 12.1$\pm$3.23 $\mu$M, respectively. The permeability-surface area product ($PS{influx}$) was calculated to be 0.0157 ml/min/10^6$ cells. $V_{max}$ and $K_m$ values for Caco-2 cells were calculated to be 6.22$\pm$0.930 pmol/min/10^6$ cells and 82.8$\pm$8.37 $\mu$M, respectively, being comparable with those of colonocytes (6.04$\pm$1.03 pmol/min/10^6$ cells and 87.8$\pm$13.2 $\mu$M, respectively). $PS_{influx}$ values for Caco-2 cells and colonocytes were calculated to be 0.0751 $\mu$l/min/10^6$ cells and 0.0688 $\mu$l/min/10^6$ cells, respectively. The more pronounced uptake of Pz-peptide by hepatocytes, when compared with Caco-2 cells and colonocytes, is probably due to its specific transporter. In conclusion, Pz-peptide, a paracellularly transported pentapeptide in the intestine and ocular epithelia, was uptaked into hepatocytes extensively. Although Pz-peptide is able to be uptaked into the Caco-2 cells and colonocytes, it is less pronounced when compared with hepatocytes. $PS_{influx}$ values of Caco-2 cells and colonocytes for unbound Pz-peptide under linear conditions were less than 0.4% when compared with that of hepatocytes.
During spawning season of female in a Korean oily bitterling, Acheilognathus koreensis, the ventral region near the base of the pectoral fin becomes to be protruded outward of body and enlarged. This ventral process consists of both organs as rectum (vent) and inner ovipositor. The rectum consists of mucosa, lamina propria-submucosa, muscularis, and squamous epithelial layer (peritoneum=serosa) surrounding them. The mucosa contains numerous mucous cells meaning acid mucopolysaccharides in nature. The inner ovipositor is similar to that of the rectum, but the mucosa have no mucous cell, unlike that of the rectum. Whereas, the outer ovipositor has a straight and long tube which are not connected with the ventral process any more. The outer ovipositor was similar to the structure of the inner ovipositor in the ventral process. However, the outer ovipositor has no muscularis, and consists of three layers: mucosa, lamina propria-submucoa, and squamous epithelia. The outer ovipositor without the muscularis seems serves as a tube that eggs discharged from the outer ovipositor allow to send inside mussel, unlike that of the inner ovipositor performing rhythmic contractions of the layers of the muscularies for propelling to the matured oocytes to the outer ovipositor.
To identify and evaluate the dichlorobenzidine(DCB)-DNA adducts in liver cell and bladder epithelial cells by $^{32}$ P-postlabeling and GC/MS-SIM, we orally exposed the dichlorobenzidine(20mg/kh body wt./day) to male Sprague-Dawley rats(l85$\pm$10g) for 14 days. Two kinds of DCB-DNA adduct(A1 and A2) were found at the same site of thin layer chromatogram of $^{32}$ P-postlabeling method in liver cells and bladder epithelial cells. In liver cells, relative adduct labeling(RAL) $\times$ 10$^{12}$ of DCB-DNA adduct A1 were 34.1$\pm$3.71 and 69.9$\pm$5.02, that of adduct A2 were 74.1$\pm$10.1 and 105.1$\pm$10.1 on 10 and 14 days after treatment, respectively. And in bladder epithelia cells, RAL $\times$ 10$^{12}$ of DCB-DNA adduct A1 were 5.92$\pm$1.60 and 15.9$\pm$1.31, that of adduct A2 were 9.81$\pm$2.81 and 22.8$\pm$1.79 on 10 and 14 days after treatment, respectively. DCB metabolites formed DNA adducts were monoacetyl-dichlorobenzidine(acDCB) and diacetyl-dichlorobenzidine(di-acDCB), which was identify by gas chromatography/mass spectrometry-scan ionization mode(GC/MS-SIM), after hydrolysis of DCB-DNA adducts isolated from live cells and bladder epithelial cells. The base peak of acDCB were 252 and 294 m/z, and that of di-acDCB were 252, 294 and 336 m/z. In conclusion, the exposed DCB formed two kinds of DCB-DNA adduct, the proximate materials of that were acDCB and di-acDCB in liver and bladder epithelial cells. And the above GC/MS-SIM method was found the DCB-DNA adducts could be monitoring by gas chromatography.
An, Chul Min;Kim, Jae Won;Jin, Young Guk;Park, Jung Jun;Lee, Jung Sick
Korean Journal of Ichthyology
/
v.19
no.3
/
pp.201-209
/
2007
The fine structures and histochemical features on the integumentary system of the grass puffer, Takifugu niphobles were examined by means of the light and transmission electron microscopy. Integumentary surface of the grass puffer showed irregular folds in light microscope. The folds of the ventral region are more pronounced than those in the dorsal region. Integumentary system is composed of outer epidermal layer and inner dermal layer. The stratified epidermal layer consists of epithelia, mucous cells, club cells, granular cells and multivacuolar gland. Epithelial cells are classified into superficial, intermediated and basal cell, and free surface of superficial cell is covered with microridges. Glands of the epidermal layer are divided into unicellular and multicellular gland. Mucous cells of multicellular gland contains mucosal materials of neutral glycoprotein. Multivacuolar gland is composed of numerous vacuole cells of about $20{\mu}m$ in axial diameter. Vacuole cells contains a large central vacuole and are connected to another by many desmosomes. The mucous glands and multivacuolar glands are more abundant in ventral region than dorsal integument. The thickness of dermis is more three to five times than epidermis in ventral integument. The collagen fibers, fibrocytes, nerve cells, basal plate of spine and chromatophore are observed in the dermal layer of compact connective tissue.
Kim, Soo-Jin;Nahm, Heun-Woo;Lee, Joon-Sang;Joo, Kyung-Whan
Applied Microscopy
/
v.30
no.1
/
pp.101-111
/
2000
In this study, the distribution of lectin receptors in Paragonimus westermani tissue was explored using colloidal gold label complexed with lectin WGA purified from wheat germ (Triticum vulgare). The lectin WGA gold complex, shown to recognize GlcNAc (N-acetylgalactosamine) and NeuNAc (N-acetylneuraminic acid) regions, was applied to detect binding sites in Lowicryl HM 20 sections viewed under electron microscope. Labeled sections of the metacercaria revealed gold particles specifically distributed on the tegumental syncytium and lamella of the excretory canal. Labeling of young adult tissue was then quantified and compared to that of adult worm tissue. Adult worm tissue sections resulted in specific gold particle distribution on the lamella of caecal epithelium and excretory canal. These results indicate that lectin WGA receptors are located in the tegumental syncytium and lamella of the excretory canal of the metacercariae, and in the lamella of the caecum and excretory canal of the young adult and adult. Therefore, the GlcNAc and NeuNAc regions in the tegumental syncytium appear to be functionally associated with cell-recognition and protection from the immune system of the host, and linked with membrane transport and absorption of nutrients in the lamella of the excretaory canal and caecal epithelia.
Objective : The etiology of chronic prostatitis is likely multifactorial, resulting from either a cascade of events after an initiating factor or from a variety of etiologic mechanisms. There is substantiating evidence to support the role of the inflammatory responses in its pathogenesis, and the clinical value in the evaluation of therapeutic efficacy. Forsythiae Frucus has been traditionally used in treatment of inflammatory diseases, including of prostatitis and urinary tract inflammation. In this study, we investigated the effects of Forsythiae Frucus on inflammatory cytokines and cyto-pathological alternation in the rat model of chronic non-bacterial prostatitis induced by castration and $17{\beta}$-estradiol treatment. Methods : Two-month-old rats were treated with $17{\beta}$-estradiol after castration for induction of experimental non-bacterial prostatitis. which is similar to human chronic prostatitis in histopathological profiles. Forsythiae Frucus as an experimental specimen, and testosterone as a positive control, were administered orally. The prostates were evaluated by histopathologlcal parameters including the epithelial score and epithelio-stromal ratio for glandular damage. and the expression of inflammatory cytokine genes including interleukin (IL)-$1{\beta}$, IL-5, IL-12, tumor necrosis factor (TNF)-$\alpha$. eotaxin, inducible nitric oxide synthase (iNOS) and cyclooxygenase-2(cox-2). Results : While prostates of control rats revealed severe acinar gland atrophy and stromal proliferation. the rats treated with Forsythiae Frucus showed a diminished range of tissue damage. Epithelial score was improved in the Forsythiae Frucus group over that of the control (P<0.05). The epithelia-stromal ratio was lower in the Forsythiae Frucus group when compared to that of the control (P<0.05). In the reverse transcription-polymerase chain reaction (RT-PCR) of inflammatory cytosine genes. Forsythiae Frucus inhibited the expression of IL-$1{\beta}$, TNF-$\alpha$, iNOS, cox-2 genes, while it modulated the expression of IL-5, which is an anti-inflammatory cytokine. Conclusions : These findings suggest that Forsythiae Frucus may protect the glandular epithelial cells and also inhibit stromal proliferation in association with the immune modulation including the suppression of inflammatory cytokines and increase of anti-inflammatory cytokines. From theses results. we suggest that Forsythiae Frucus could be a useful remedy agents for treating chronic non-bacterial prostatitis.
Objective : Although chronic non-bacterial prostatitis is a common disease, it is very difficult to treat effectively. Lygodium japonicum has been traditionally used in treatment of urinary tract inflammation and voiding disturbance. In this study, we investigated the therapeutic effects and action mechanism of Lygodium japonicum in the rat model of non-bacterial prostatitis induced by castration and testosterone treatment. Methods : Five-month-old rats were treated with 17$\beta$-estradiol after castration for induction of experimental non-bacterial prostatitis, which is similar to human chronic prostatitis in histopathological profiles. Lygodium japonicum and testosterone were administered as an experimental specimen and a positive control. respectively. The prostates were evaluated by histopathological parameters including the epithelial score and epithelio-stromal ratio for glandular damage. PCNA labeling index for cyto-proliferation and a TUNEL(deoxyuridine triphosphate biotin nick end-labeling) assay for cell apoptosis. Results : While prostates of control rats revealed severe acinar gland atrophy and stromal proliferation, the rats treated with Lygodium japonicum showed a diminished range of tissue damage. Epithelial score was improved in the Lygodium japonicum group over that of the control (P<0.05). The epithelio-stromal ratio was lower in the Lygodiutn japonicum group when compared to that of the control (P<0.05). Although there was no difference in PCNA and TUNEL positive cells of the glandular epithelia. we found an decreased number of PCNA positive cell and concurrent increase of TUNEL positive cells in the stroma of Lygodium japonicum treated rats (P<0.01). Conclusions : These findings suggest that Lygodium japonicum may protect the glandular epithelial cells and also inhibit stromal proliferation in association with suppression of cyto-proliferation and stimulation of apoptosis. We concluded that Lygodium japonicum could be a useful remedy agents for treating chronic non-bacterial prostatitis.
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