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Expression of obese gene and leptin secretion in the human placenta: The possibility of leptin as a novel placenta-derived hormone
A.10
THE ROLE OF MATRIX METALLOPROTEINASE TISSUE INHIBITOR OF (MMP), METALLOPROTEINASE (TIMP), AND TRANSFORMING GROWTH FACTOR-j 1 (TGF- j 11, IN PREECLAMPSIA. S. Matsushita, H. Narumiya, M. Hirata, H. Furuya, H. Yabushita, M. Noguchi and M. Nakanishi, Department of Obstetrics and Gynecology, Aichi Medical University, Aichi, Japan. In order to study the role of matrix metalloproteinase-2, -9 (MMP-2, -9), tissue inhibitor of metalloproteinase-I, -2 (TIMP-1, -2) and its regulation in preeclampsia, the expression of MMP-2, MMP-9, TIMP-1, TIMP-2 and TGF-jl in preeclampsia was examined by EIA, zymographic analysis and immunohistochemistry. Two groups were studied; control healthy pregnant women (n=40), and preeclamptic patients (n=lO), Serum TIMP- 1 and TGF-/?l levels in preeclampsia were significantly higher than in the control group. Zymographic analysis using placental tissue showed that both MMP-2 and MMP-9 activities were increased in preeclampsia. Immunohistochemical localization of MMP-2, TIMP-2 and TGF-11 was noted in both syncytiotrophoblasts and the stromal compartment, and that of MMP-9 and TIMP-1 was noted only in stromal compartment. However, there was no difference in the frequency of localization between the two groups. These data suggested that MMP-2, MMP-9, TIMP-1 and TGF-11 play an important role in preeclampsia.
CHANGE OF MEMBRANE TYPE-MATRIX METALLOPROTEINASE GENE EXPRESSION IN HUMAN ENDOMETRIUM THROUGH THE MENSTRUAL CYCLE -ANALYSIS BY QUANTITATIVE RT-PCR AND IN SITU HYBRIDAIZATIONM.Nakano, T.Hara, T.Hayama and K.Ohama Department of Obstetrics and Gynecology, Hiroshima University school of medicine, Japan Most matrix metalloproteinases(MMP) are thought to be implicated in remodeling of the human endometrium through the menstrual cycle and implantation. However, the change of membrane type-MMP(MT-MMP) has not been investigated in human endometrium. We quantified gene expression of MT-MMP and MMP-7 by RT-PCR and investigated the pattern of MT-MMP gene expression by in situ hybridization using a Digoxigenin-labeled RNA probe that was synthesized from MT-MMP PCR products. The highest level of MMP-7 mRNA was detected in all samples from the menstrual phase and the lowest level from the secretory phase. In contrast, for MT-MMP, the expression level was the highest in the secretory phase and lowest level in the proliferative phase. The hybridization signal of MT-MMP gene were also stronger in the secretory phase than that in the proliferative phase. We suggest that the expression of MT-MMP gene might be regulated by a mechanism which differs from that for the MMP-7 gene.
Placenta EXPRESSION OF OBESE GENE AND LEPTIN SECRETION IN THE HUMAN PLACENTA: THE POSSIBILITY OF LEPTIN AS A NOVEL PLACENTADERIVED HORMONE. H. Mise, N. Sagawa, T. Matsumoto, S. Yura, B. Liu. *H. Masuzaki, *Y. Oaawa, *K. Hosoda, *K. Nakao. Department of Obstetrics-and Gynecology, and *Department of Clinical Science and Medicine, Kyoto University Graduate School of Medicine, Kyoto, Japan. Objtutives: We previously reported that plasma @tin levels in pregnant women are significantly higher than those of body mass index (BMI)match& non-pregnant women and that theelevated leptin in the maternal plasma at least partlyoriginates from placental trophoblast. In the present study, we measured plasma leptin levels in pregnant women and umbilicalcord of newlxnns to elucidate the metabolism and significance of leptin in human pregnancy. In addition, the expression of obese gene and localization of leptin in intrauterine tissues were studied. Subjects and Methods: Blcod specimens were drawn from these antccubital vein of 87 normal non-oresnant women and 78 normal oresnant women (25 36 wks). Plasma levels were determined by a ridioimnntnoassay specific for human leptin (Hosoda et al, BBRC 221: 234, 1996). Localization of leptin was examined by an indirect immunofluorescence method using anti-human leptin antibody. Obese gene expression was examined by Northern blot analy$.s. Results Plasma lepiin levels in normal pregnant women were 35.0 2 2.9 rig/ml (meanfSEM, n=78), which was 2-fold higher than that of normal non-pregnant women. The elevated plasma leptin levels decreased rapidly after delivery of the placenta. Leptin concentrations in the umbilical arterial plasma (mean 9.9 @ml, n=36) were approximately one third of those in the paired maternal plasma, but were significantly lower than those in the paired umbilical venous plasma (13.4 q/ml, n=36. ~0.01). Indirect immunofluorescexe for leptin detected an intense fluorescence in syncytiotrophoblast cells. Northern blot analysis detected ob mRNA in the chorionic villous tissue and amnion. Conclusions: The present study indicates that the obese gene is expressed in the human placenta and leptin is seeretcd into both maternal and fetal circulations.
ie&
TELOMERASE ACTIVITY IN CHORIONIC VILLI AND TROPHOBLASTIC DISEASE. H.Nishi(l), K.Isaka(l), K.Shiraishi(l), N.Yahata(a), KToyama(2) and M.Takayama(l), Department of Obstetrics and Gynecology(l) and 1st. Department of Internal Medicine(2), Tokyo Medical College, Tokyo, Japan. The ribonucleoprotein, telomerase may play an important role in ceil immortality by catalyzing the de ~IUVO synthesis of telomere ends. Telomerase activity has been observed in immortal cells and most cancer cells but nor in normal somatic cells with the exception of cells such as germ line and stem cells. Recently, the telomeric repeat amplification protocol assay (TRAP), a sensitive method of determining telomerase activity based on PCR has become available. Some reports have been made concerning telomerase activity detected in various gynecological tumors using this method. We have made a study comparing the quantitated telomerase activity in tissues obtained from 16 normal chorionic villi, 7 hydatidiform moles;(partial mole, total mole and invasive mole) and 2 choriocarcinomas using the TRAP assay in collaboration with the internal telomerase assay standard(ITAS) and an automatic DNA sequencer. The results revealed telomerase activity of 1.181fi2.86 in normal chorionic villi with a decrease in activity accordance with gestational age and a slightly elevated telomerase activity of 3.17k2.81 in moles and 1.35kO.57 in choriocarcinomas. This suggests the relation of telomerase activity to the regulation of trophoblastic cell proliferation. Therefore, although telomerase activity is seen in many villi, an elevation can be indicative of trophoblastic diseases.
(1997),
Vol. 18
THE ROLE OF MATRIX METALLOPROTEINASE TISSUE INHIBITOR OF (MMP), METALLOPROTEINASE (TIMP), AND TRANSFORMING GROWTH FACTOR-j 1 (TGF- j 11, IN PREECLAMPSIA. S. Matsushita, H. Narumiya, M. Hirata, H. Furuya, H. Yabushita, M. Noguchi and M. Nakanishi, Department of Obstetrics and Gynecology, Aichi Medical University, Aichi, Japan. In order to study the role of matrix metalloproteinase-2, -9 (MMP-2, -9), tissue inhibitor of metalloproteinase-I, -2 (TIMP-1, -2) and its regulation in preeclampsia, the expression of MMP-2, MMP-9, TIMP-1, TIMP-2 and TGF-jl in preeclampsia was examined by EIA, zymographic analysis and immunohistochemistry. Two groups were studied; control healthy pregnant women (n=40), and preeclamptic patients (n=lO), Serum TIMP- 1 and TGF-/?l levels in preeclampsia were significantly higher than in the control group. Zymographic analysis using placental tissue showed that both MMP-2 and MMP-9 activities were increased in preeclampsia. Immunohistochemical localization of MMP-2, TIMP-2 and TGF-11 was noted in both syncytiotrophoblasts and the stromal compartment, and that of MMP-9 and TIMP-1 was noted only in stromal compartment. However, there was no difference in the frequency of localization between the two groups. These data suggested that MMP-2, MMP-9, TIMP-1 and TGF-11 play an important role in preeclampsia.
CHANGE OF MEMBRANE TYPE-MATRIX METALLOPROTEINASE GENE EXPRESSION IN HUMAN ENDOMETRIUM THROUGH THE MENSTRUAL CYCLE -ANALYSIS BY QUANTITATIVE RT-PCR AND IN SITU HYBRIDAIZATIONM.Nakano, T.Hara, T.Hayama and K.Ohama Department of Obstetrics and Gynecology, Hiroshima University school of medicine, Japan Most matrix metalloproteinases(MMP) are thought to be implicated in remodeling of the human endometrium through the menstrual cycle and implantation. However, the change of membrane type-MMP(MT-MMP) has not been investigated in human endometrium. We quantified gene expression of MT-MMP and MMP-7 by RT-PCR and investigated the pattern of MT-MMP gene expression by in situ hybridization using a Digoxigenin-labeled RNA probe that was synthesized from MT-MMP PCR products. The highest level of MMP-7 mRNA was detected in all samples from the menstrual phase and the lowest level from the secretory phase. In contrast, for MT-MMP, the expression level was the highest in the secretory phase and lowest level in the proliferative phase. The hybridization signal of MT-MMP gene were also stronger in the secretory phase than that in the proliferative phase. We suggest that the expression of MT-MMP gene might be regulated by a mechanism which differs from that for the MMP-7 gene.
Placenta EXPRESSION OF OBESE GENE AND LEPTIN SECRETION IN THE HUMAN PLACENTA: THE POSSIBILITY OF LEPTIN AS A NOVEL PLACENTADERIVED HORMONE. H. Mise, N. Sagawa, T. Matsumoto, S. Yura, B. Liu. *H. Masuzaki, *Y. Oaawa, *K. Hosoda, *K. Nakao. Department of Obstetrics-and Gynecology, and *Department of Clinical Science and Medicine, Kyoto University Graduate School of Medicine, Kyoto, Japan. Objtutives: We previously reported that plasma @tin levels in pregnant women are significantly higher than those of body mass index (BMI)match& non-pregnant women and that theelevated leptin in the maternal plasma at least partlyoriginates from placental trophoblast. In the present study, we measured plasma leptin levels in pregnant women and umbilicalcord of newlxnns to elucidate the metabolism and significance of leptin in human pregnancy. In addition, the expression of obese gene and localization of leptin in intrauterine tissues were studied. Subjects and Methods: Blcod specimens were drawn from these antccubital vein of 87 normal non-oresnant women and 78 normal oresnant women (25 36 wks). Plasma levels were determined by a ridioimnntnoassay specific for human leptin (Hosoda et al, BBRC 221: 234, 1996). Localization of leptin was examined by an indirect immunofluorescence method using anti-human leptin antibody. Obese gene expression was examined by Northern blot analy$.s. Results Plasma lepiin levels in normal pregnant women were 35.0 2 2.9 rig/ml (meanfSEM, n=78), which was 2-fold higher than that of normal non-pregnant women. The elevated plasma leptin levels decreased rapidly after delivery of the placenta. Leptin concentrations in the umbilical arterial plasma (mean 9.9 @ml, n=36) were approximately one third of those in the paired maternal plasma, but were significantly lower than those in the paired umbilical venous plasma (13.4 q/ml, n=36. ~0.01). Indirect immunofluorescexe for leptin detected an intense fluorescence in syncytiotrophoblast cells. Northern blot analysis detected ob mRNA in the chorionic villous tissue and amnion. Conclusions: The present study indicates that the obese gene is expressed in the human placenta and leptin is seeretcd into both maternal and fetal circulations.
ie&
TELOMERASE ACTIVITY IN CHORIONIC VILLI AND TROPHOBLASTIC DISEASE. H.Nishi(l), K.Isaka(l), K.Shiraishi(l), N.Yahata(a), KToyama(2) and M.Takayama(l), Department of Obstetrics and Gynecology(l) and 1st. Department of Internal Medicine(2), Tokyo Medical College, Tokyo, Japan. The ribonucleoprotein, telomerase may play an important role in ceil immortality by catalyzing the de ~IUVO synthesis of telomere ends. Telomerase activity has been observed in immortal cells and most cancer cells but nor in normal somatic cells with the exception of cells such as germ line and stem cells. Recently, the telomeric repeat amplification protocol assay (TRAP), a sensitive method of determining telomerase activity based on PCR has become available. Some reports have been made concerning telomerase activity detected in various gynecological tumors using this method. We have made a study comparing the quantitated telomerase activity in tissues obtained from 16 normal chorionic villi, 7 hydatidiform moles;(partial mole, total mole and invasive mole) and 2 choriocarcinomas using the TRAP assay in collaboration with the internal telomerase assay standard(ITAS) and an automatic DNA sequencer. The results revealed telomerase activity of 1.181fi2.86 in normal chorionic villi with a decrease in activity accordance with gestational age and a slightly elevated telomerase activity of 3.17k2.81 in moles and 1.35kO.57 in choriocarcinomas. This suggests the relation of telomerase activity to the regulation of trophoblastic cell proliferation. Therefore, although telomerase activity is seen in many villi, an elevation can be indicative of trophoblastic diseases.
(1997),
Vol. 18