Keratinocyte growth factor (KGF) functions in epithelial growth and differentiation in many tissues and organs. KGF is expressed in the uterine endometrial epithelial cells during the estrous cycle and pregnancy in pigs, and receptors for KGF (KGFR) are expressed by conceptus trophectoderm and endometrial epithelia. KGF has been shown to stimulate the proliferation and differentiation of conceptus trophectoderm. However, the role of KGF on the endometrial epithelial cells has not been determined. Therefore, this study determined the effect of KGF on proliferation and differentiation of endometrial epithelial cells in vitro and in vivo using an immortalized porcine luminal epithelial (pLE) cell line and KGF infusion into the uterine lumen of pigs between Days 9 and 12 of estrous cycle. Results showed that KGF did not stimulate proliferation of uterine endometrial epithelial cells in vitro and in vivo determined by the $^3$H]thymidine incorporation assay and the proliferating cell nuclear antigen staining, respectively. Effects of KGF on expression of several markers for epithelial cell differentiation, including integrin receptor subunits $\alpha$4, $\alpha$5 and $\beta$1, plasmin/trypsin inhibitor, uteroferrin and retinol-binding protein were determined by RT-PCR, Northern and slot blot analyses, and immunohistochemisty, and KGF did not affect epithelial cell differentiation in vitro and in vivo. These results show that KGF does not induce epithelial cell proliferation and differentiation, suggesting that KGF produced by endometrial epithelial cells acts on conceptus trophectoderm in a paracrine manner rather than on endometrial epithelial cells in an autocrine manner.
BACKGROUND/OBJECTIVES: This study was conducted to assess the potential of St. John's Wort (Hypericum perforatum) to prevent obesity and abnormalities in lipid metabolism induced by ovariectomy in a rat model without stimulatory activity on uterus. MATERIALS/METHODS: Ovariectomized (OVX) rats were treated for 6 weeks with 70% ethanol extracts of Hypericum perforatum [HPEs: whole plant (WHPE) and flower and leaves (FLHPE)], ${\beta}$-estradiol-3-benzoate at a dose of $50{\mu}g/kg/day$ (E2) or vehicle (distilled water). RESULTS: As expected, OVX increased body weight gain and adiposity and showed higher food efficacy ratio. OVX also increased the serum cholesterol as well as insulin resistance, while reducing uterus weight and uterine epithelial proliferation rate. HPEs (WHPE and FLHPE) showed estrogen-like effect on body weight gain, adipose tissue weight and food efficacy ratio in OVX rats. HPEs prevented hypercholesterolemia induced by OVX more effectively than E2. E2 increased uterus weight and epithelial proliferation rate in OVX rats, while HPEs maintained them at the level of the sham-operated animals. CONCLUSIONS: Our finding demonstrates that HPEs can be considered as an effective agent to prevent OVX-induced obesity without stimulatory activity on uterus.
Uterine cells carry out proliferation and differentiation for preparation the embryonic implantation during pregnancy. Therefore regulation of the cell proliferation is an essential step for uterine preparation, but there is not much information about the proliferation related genes in pregnant uterus. To identify these implantation specific genes, a PCR-select cDNA subtraction method was employed and got a few genes. One of the identified genes is a novel gene encoding oral tumor suppressor doc-1. To detect the doc-1 expression on the pregnant uterus, dot blotting, RT-PCR, and in situ hybridization were employed. Dot blotting revealed that doc-1 mRNA expression increase after implantation. During normal pregnancy, doc-1 mRNA expression was detected as early as day 1 of pregnancy with RT-PCR. Its expression was increased about 15 times after embryonic implantation. doc-1 transcript was localized in luminal epithelial cells but it was very faint during preimplantation. After starting the implantation, it localized in the stromal cells; heightened expression of doc-1 correlates with intense stromal cell proliferation surrounding the implanting blastocyst on day 6 morning. However in the decidualized cells, the intensity of localized doc-1 mRNA was weak. From those results, it is revealed that doc-1 express at pregnant uterus of the mouse. In addition it is suggested that doc-1 is the gene regulating the proliferation of the luminal epithelial cells and stromal cells during early implantation and decidualization.
Kim, Juhye;Cha, Sunyeong;Lee, Min Young;Hwang, Yeon Jeong;Yang, Eunhyeok;Choi, Donchan;Lee, Sung-Ho;Cheon, Yong-Pil
Development and Reproduction
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v.23
no.3
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pp.263-275
/
2019
Based on our preliminary results, we examined the possible role of low-dose and chronic-exposing of the chemicals those are known as endocrine disrupting chemical (EDC), on the proliferation of uterine endometrium and the localization of steroid receptors. Immunohistochemical or immunofluorochemical methodology were employed to evaluate the localization of antigen identified by monoclonal antibody Ki 67 protein (MKI67), estrogen receptor 1 (ESR1), estrogen receptor 2 (ESR2), and progesterone receptor (PGR). In $133{\mu}g/L$ and $1,330{\mu}g/L$ di(2-ethylhexyl) phthalate (DEHP) and $50{\mu}g/L$ nonylphenol (NP) groups, the ratio of MKI67 positive stromal cells was significantly increased but not in $500{\mu}g/L$ NP group. The ratios of MKI67 positive glandular and luminal epithelial cells were also changed by the chronic administration of NP and DEHP in tissue with dose specific manner. ESR1 signals were localized in nucleus in glandular and luminal epithelia of control group but its localization was mainly in cytoplasm in DEHP and NP administered groups. On the other hand, it was decreased at nucleus of stromal cells in $1,330{\mu}g/L$ DEHP group. The colocalization patterns of these nuclear receptors were also modified by the administration of these chemicals. Such a tissue specific and dose specific localization of ESR2 and PGR were detected as ESR1 in all the uterine endometrial tissues. These results show that the chronic lows-dose exposing of NP or DEHP modify the localization and colocalization of ESRs and PGR, and of the proliferation patterns of the endometrial tissues.
Im, Ji Woo;Lee, Chae Young;Kim, Dong-Hwan;Bae, Hae-Rahn
Development and Reproduction
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v.24
no.3
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pp.177-185
/
2020
Although many aquaporin (AQP) transcripts have been demonstrated to express in the female reproductive tract, the defined localizations and functions of AQP subtype proteins remain unclear. In this study, we investigated the expression of AQP1, AQP3, AQP5, AQP6, and AQP9 proteins in female reproductive tract of mouse and characterized their precise localizations at the cellular and subcellular levels. Immunofluorescence analyses for AQP1, AQP3, AQP6, and AQP9 showed that these proteins were abundantly expressed in female reproductive tract and that intense immunoreactivities were observed in mucosa epithelial cells with a subtype-specific pattern. The most abundant aquaporin in both vagina and uterine cervix was AQP3. Each of AQP1, AQP3, AQP6, and AQP9 exhibited its distinct distribution in stratified squamous or columnar epithelial cells. AQP9 expression was predominant in oviduct and ovary. AQP1, AQP3, AQP6, and AQP9 proteins were mostly seen in apical membrane of ciliated epithelial cells of the oviduct as well as in both granulosa and theca cells of ovarian follicles. Most of AQP subtypes were also expressed in surface epithelial cells and glandular cells of endometrium in the uterus, but their expression levels were relatively lower than those observed in the vagina, uterine cervix, oviduct and ovary. This is the first study to investigate the expression and localization of 5 AQP subtype proteins simultaneously in female reproductive tract of mouse. Our results suggest that AQP subtypes work together to transport water and glycerol efficiently across the mucosa epithelia for lubrication, proliferation, energy metabolism and pH regulation in female reproductive tract.
Objective: Zearalenone (ZEA) has estrogen-like effects. Our previous study has shown that ZEA (0.5 to 1.5 mg/kg) could induce abnormal uterine proliferation through transforming growth factor signaling pathway. To further study the other regulatory networks of uterine hypertrophy caused by ZEA, the potential mechanism of ZEA on porcine endometrial epithelial cells (PECs) was explored by the Illumina Hiseq 2000 sequencing system. Methods: The PECs were treated with ZEA at 0 (ZEA0), 5 (ZEA5), 20 (ZEA20), and 80 (ZEA80) µmol/L for 24 h. The collected cells were subjected to cell cycle, RNA-seq, real-time quantitative polymerase chain reaction, immunofluorescence, and western blot analysis. Results: The proportion of cells in the S and G2 phases decreased (p<0.05), but the proportion of cells in the G1 phase increased (p<0.05) in the ZEA80 treatment. Data analysis revealed that the expression of Wnt pathway-related genes, estrogen-related genes, and mitogen-activated protein kinase pathway-related genes increased (p<0.05), but the expression of genetic stability genes decreased (p<0.05) with increasing ZEA concentrations. The relative mRNA and protein expression of WNT1, β-catenin, glycogen synthase kinase 3β (GSK-3β) were increased (p<0.05) with ZEA increasing, while the relative mRNA and protein expression of cyclin D1 (CCND1) was decreased (p<0.05). Moreover, our immunofluorescence results indicate that β-catenin accumulated around the nucleus from the cell membrane and cytoplasm with increasing ZEA concentrations. Conclusion: In summary, ZEA can activate the Wnt/β-catenin signaling pathway by up-regulating WNT1 and β-catenin expression, to promote the proliferation and development of PECs. At the same time, the up-regulation of GSK-3β and down-regulation of CCND1, as well as the mRNA expression of other pathway related genes indicated that other potential effects of ZEA on the uterine development need further study.
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, Yeo-Yae;Lee, Dong-Nyung;Seo, Il-Bock;Kim, Hyung-Jun
The Journal of Korean Obstetrics and Gynecology
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v.19
no.2
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pp.162-185
/
2006
Purpose : This study was carried out to investigate the effects of Gamigwichulpajing-Tang(GGT) on the development of experimentally-induced endometriosis in rats. Materials and Methods : Endometriosis was induced in rats by auotransplanting 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 GGT treated group. GGT(700mg/head) was orally administrated for 15days after operation. Then we measured the body weight, the volumes of endometriotic implants, the weight of uterus and ovary, and investigated the content of cytokines(MCP-1, $TNF-{\alpha}$, $IL-l{\beta}$) in serum and ascites. Histopathology, immunohistochemistry for COX-2, and histochemistry for mast cell in transplanted uterine tissue were performed. Results :- The $volume(mm^3)$ of endometriotic implants in GGT-treated group$(53.50\;{\pm}18.63)$ was significantly decreased(p<<0.01) compared with control group$(404.50{\pm}317.68)$. - The content(pg/ml) of MCP-1 in ascites in GGT-treated group$(4265{\pm}108)$ was significantly decreased(p<<0.001) compared with control group$(8632{\pm}1245)$. - The content(pg/ml) of $TNF-{\alpha}$ in serum in GGT-treated group$(64.5{\pm}21.6)$ was significantly decreased(p<<0.05) compared with control group$(147.1{\pm}78.2)$. - The content(pg/ml)- of $TNF-{\alpha}$ in ascites in GGT-treated group$(738.3{\pm}502.4)$ was significantly decreased(p<<0.05) compared with control group$(1245.2{\pm}362.2)$. - The percentage(%) of positive epithelial layers for COX-2 in GGT-treated group$(22.9{\pm}9.3)$ was significantly decreased(p<<0.001) compared with control group$(50.2{\pm}8.2)$. - The number of mast cells in adjacent tissue of transplanted uterine tissue in GGT-treated group$(61.4{\pm}13.9)$ was significantly decreased(p<<0.001) compared with control group$(109.3{\pm}30.2)$. - The number of mast cells in stroma of transplanted uterine tissue in GGT-treated group$(9.4{\pm}2.7)$ was significantly decreased(p<<0.001) compared with control group$(26.0{\pm}7.7)$. - Histopathologically, proliferation of endomeuiotic epithelia and stroma, and infiltration of inflammatory cells in transplanted uterine tissue of GGT-treated group were weakly observed than those of control group. Conclusion :On the basis of these results, we concluded that Gamigwichulpajing-Tang have inhibiting effects on the development of transplanted uterine tissue. And these effects may be related with decreased Production of MCP-1 and $TNF-{\alpha}$, decreased expression of COX-2, and decreased infiltration of mast cells by administration of Gamigwichulpajing-Tang.
Kim, Chul-Soo;Lee, Dong-Nyung;Seo, Il-Bock;Kim, Hyung-Jun
The Journal of Korean Obstetrics and Gynecology
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v.20
no.1
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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}$, decreased expression of COX-2, and decreased infiltration of mast cells by administration of Keukhachukeo-Tang.
Purpose: This study was performed to investigate the effects of Dangguijakyaksan on the development of experimentally-induced endometriosis in rats. Methods: Endometriosis was induced in rats by autotransplanting uterine tissue to the peritoneum and divided them into three groups: (1) sham-operated group(n=8). (2) surgically induced endometriosis and untreated control group(n=8). (3) surgically induced endometriosis and Dangguijakyaksan treated group. Dangguijakyaksan was orally administrated for 15 days after operation. Then we measured the body weight. the volume of endometriotic implants. the weight of uterus and ovary. and analysed the concentration of cytokines (MCP-1, TNF-$\alpha$, IL-1$\beta$, IL-6) in peritoneal fluids. Histopathology, immunohistochemistry for COX-2 and VEGF. and histochemistry for mast cell in transplanted uterine tissue were also performed. Results: - The volume($mm^3$) of endometriotic implants in Dangguijakyaksan treated group was significantly decreased compared with control group. - The concentration(pg/ml) of MCP-1, TNF-$\alpha$ and IL-l$\alpha$ in peritoneal fluids in Dangguijakyaksan treated group was significantly decreased compared with control group. - Histopathologically, proliferation of endometriotic epithelia. infiltration of inflammatory cells and angiogenesis in transplanted uterine tissue of Dangguijakyaksan treated group were weakly observed than those of control group. - The percentage of positive epithelial layers for COX-2 in Dangguijakyaksan treated group was significantly decreased compared with control group. - The VEGF expression of endometriotic epithelia, neovascular endothelia and stromal cells in transplanted uterine tissue of Dangguijakyaksan treated group were weakly observed than those of control group. - The number of mast cells in transplanted uterine tissue in Dangguijakyaksan treated group was significantly decreased compared with control group. Conclusion: On the basis of these results. we concluded that Dangguijakyaksan has inhibiting effects on the development of transplanted uterine tissue. And these effects may be related with decreased production of MCP-1, TNF-$\alpha$ and IL-1$\beta$, and decreased expression of COX-2 and VEGF, and inhibition of mast cell infiltration by administration of Dangguijakyaksan.
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