| Background:The concept of Gestational diabetes mellitus(GDM)has been put forward for about half century years.And a study shows that the pathogenesis of GDM is closely related with insulin resistance, fat cells, inflammatory factors, genetic factors and immune factors. Leptin(LEP), one of fat cells, is proved to be a key factor involved in sugar and fat metabolism. Since it’s found, by many scholars in China and abroad.The function of leptin biology is determined by one’s leptin concentration, receptor content and its distribution.Leptin is mainly composed of synthesis and secretion of adipose tissue, the study on metabolic disease during pregnancy shows that placental tissue is another important organization that can synthesize and secrete leptin in addition to the fat tissue and it’s also found that the placenta tissue can not only synthesis leptin but also express leptin receptor.The dysfunction of Placental leptin can lead to GDM. Although leptin receptors has different classification(a, b, c, d, e, f), and a variety of forms(long form of of leptin receptor, short form of leptin receptor and soluble leptin receptor), the leptin receptor research mainly focuses on the long form of leptin receptor(b type, Ob-R), mainly distributed on the central nervous system,and soluble leptin receptor(e type, SLR). Placental tissue can express the Ob-R while SLR, mainly existing in serum, is involved in regulating blood thin element concentration. Scholars abroad find the amount of expression of SLR in people with type 2 diabetes is lower than that with the normal glucose tolerance. Our preliminary study found SLR expressed in GDM group was lower than the pregnant women with normal glucose tolerance.Placental tissue can secrete a variety of hormones involved in the regulation of glucose metabolism. It has been proved that abnormal glucose tolerance during pregnancy pregnant women blood progesterone level was obviously higher than that of pregnant women with normal glucose tolerance. Our previous studies have found that pregnant women blood leptin levels of abnormal glucose tolerance was positively related with progesterone and progesterone is a risk factor for GDM. AS scholars in China and abroad confirmed pregnancy hormones can affect the biological function of leptin and some scholars found that the activation of matrix metalloproteinases depend on progesterone,some scholars even put forward A Disintegrin And Metalloprotease domain 10(ADAM10) by cracking long transmembrane leptin receptor(Ob-R) produce soluble leptin receptor(SLR),we make a speculation that the pregnancy test hormone can affect ADAM10 and ADAM10 can affect the synthesis of SLR by the cracking Ob-R and leptin function, which result in abnormal glucose metabolism during pregnancy and cause GDM.Objectives:Firstly,Have a better understanding of ADAM10 in the expression of placental tissue; Secondly, make an analysis of different concentration of progesterone in human villi trophoblastic cells on the express of ADMA10, Ob-R, SLR and LEP, and explore the role of progesterone, ADMA10, SLR on the pathogenesis of GDM mechanism.Methods:1. 15 cases of full-term single GDM pregnant women placenta tissue is regarded as a research team while 15 cases of full-term fetal normal pregnancy women group, a comparative group. All the people was tested 75 g Oral Glucose Tolerance during 24 to 28 pregnancy.The one conforms to the International association of diabetes and pregnancy study groups(IADPSG) diagnostic criteria of blood sugar is defined as GDM group, and the other one is the normal pregnancy group. The Western Blotting, qRT- PCR detection ADAM10 protein and m RNA will be used to analyze the expression quantity and the difference of placenta tissue in two groups.2. The embryo villus tissue from normal people with pregnancy 6-10 weeks, and the ones confirmed by ultrasonic intrauterine pregnancy voluntarily terminate pregnancy in outpatient obstetrics and gynecology hospital will be separated and cultivated a pregnancy villous trophoblast cells. S-P method is used to identify the trophoblastic cells. Four groups will be given 0 ng/ml, 100 ng/ml, 150 ng/ml, 200 ng/ml of progesterone respectively to cultivate villi pregnancy trophoblastic cells. 24 hours later, cells and the supernatant will be collected to analyze the expression of relative quantity of human villous trophoblast trophoblastic cells ADAM10, Ob-R protein by Western Blotting and ELISA is to detect the content of SLR, LEP in supernatant. By means of comparing different concentration progesterone for pregnancy villus ADAM10 trophoblast cells on the influence of the Ob-R expression, secretion SLR, and the content of LEP, the speculation ADAM10 is a related with GDM will be proved further.Results:1. The first part: the Western Blotting tests confirmed the content of ADAM10 protein in GDM group was obviously lower than normal pregnancy group(0.98 ± 0.26 VS. 1.59 ± 0.52, P < 0.01), and qRT-PCR results show ADAM10 m RNA expressed in GDM group is lower than normal pregnancy group(0.37 ± 0.25 VS. 1.00 ± 0.00, P < 0.01).2.The second part:(1)0 ng/ml,100 ng/ml,150 ng/ml and 200 ng/ml progesterone concentration relative to express human villi trophoblastic cells ADAM10 were 1.9411±制相关研究,Schaab M[10]研究证实ADAM10可通过蛋白激酶C(Protein Kinase C,PKC)0.4301, 1.3700 ± 0.3041, 0.7743 ± 0.1844, 0.2433 ± 0.6122, between two groups to compare(P < 0.05) which has a statistical significance.(2) Groups of cells express Ob-R relative expression were 0.5796 ± 0.3197, 0.7070 ± 0.3155, 1.2446 ± 0.6636, and 1.5168 ± 0.8224, the comparison between groups(P > 0.05) has no statistical significance.(3) Concentration in the supernatant are: 9.4990 ± 0.8068、7.3213 ± 0.1496、5.8720 ± 0.0180, and 3.8758 ± 1.0690(ng/ml),(P < 0.05) in groups of SLR has statistical significance.(4) The concentration in the supernatant ranging 144.87 ± 10.975, 287.09 ± 28.762, 332.42 ± 14.867, 443.09 ± 12.389(ng/ml),(P < 0.05), groups of LEP is statistically significant.Conclusions: 1. ADAM10 is involved in the expression of placenta tissue and it may participate in the pathological process of GDM on the condition that and the content of the placenta tissue ADAM10 in GDM group is lower than normal pregnancy group,. 2. Progesterone has an effect on pregnancy villous trophoblast cells.The increase of concentration of ADAM10 is associated with the decrease of SLR expression. Suggest progesterone affect ADAM10 expression and reduce SLR secretion, leptin biology function, and cause glucolipid metabolic changes.Progesterone may affect the involvement of GDM pathophysiological process. |
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