The Effect Of Insulin And Insulin-like Growth Factor System And Vascular Endothelial Growth Factor On Etiologica Mechanism Of Polycystic Ovarian Syndrome

Objective:To study the levels of Insulin(Ins), Glucose(Glu), insulin-likegrowth factor-I (IGF-1), testosterone (T) and vascular endothelial growth factor(VEGF) in serum and the levels of insulin-like growth factor-Ⅱ ( IGF-Ⅱ) andVEGF measured in follicular fluid and their relationship .Cytologiclocalization of IGF-1,VEGF, Insulin receptor (Ir),insulin-like growth factor-Ireceptor(IGF-Ir) in ovaries were measured in polycystic ovary syndrome(PCOS)and control groups,which would help to explain further the failure of folliculardevelopment regulated by local factors,such as IGFs ,VEGF,which are involvedin the pathogenesis of PCOS.Methods: GOD-POD method was used to detect the Glu in serum of 38cases PCOS and 30 cases control groups;ELISA method was used to detect theVEGF in serum and VEGF in follicular fluid of 38 cases PCOS and 30 casescontrol groups;SP immunohistochemical staining method was used to detect theprotein expression of Ir, IGF-Ir, IGF-1 and VEGF in ovaries of 38 cases PCOSand 30cases control groups;In situ hibridization was used for detecting theexpression of IGF-1 mRNA,VEGF mRNA in ovaries of 38 cases PCOS and30 cases control groups.Results:1. Observing the sufferers of PCOS and their clinical datasrelated to metabolism of glucose and lipid manifests the PCOS' sufferersexist the serious metabolic disorder of glucose and lipid.Discovering in thebelly cavity mirror and surgical operations the ovary blood vessels of PCOS'sufferer are more abundant than the normal women's.The theca-interstitialcells in PCOS were more than in control groups. 2.The levels of serumT,Ins,IGF-Ⅰa nd VEGF and the level of follicular fluid VEGF were(144.75±26.4)ng/dl, (20.99±10.84)μIU/ml,(116.11±34.48)ng/ml ,(246.35±332.9.96)pg/ml and(520.32±45.49 )pg/ml,which were significantlyhigher than those of the control groups,respectively ( 37.88±17.07 )ng/dl,(10.84±2.74)μIU/ml,(55.42±35.20)ng/ml,(85.61±32.10)pg/ml,(245.63±35.40)pg/ml( P<0.001,P<0.001,P<0.001,P<0.001,P<0.05). The follicularfluid IGF-Ⅱ level were(0.132±0.018)ng/ml, which were significantly lowerthan that of the control groups(0.198±0.035)ng/ml(P<0.001);⒊The serumIns and T level of A canthosis nigricans masc were (27.23±10.17)μIU/ml and(159.24±28.68)ng/dl ,which were significantly higher than those of the Acanthosis nigricans negative groups,respectively( 15.38±4.14)μIU/ml,(131.71±15.61)ng/dl(P<0.05, P<0.05). The serum IGF-Ⅰ level of Acanthosis nigricans masc were ( 102.99±35.869 ) ng/ml, which weresignificantly lower than that of the A canthosis nigricans negative groups(127.92±29.24)ng/ml(P<0.05);⒋Within the PCOS group Ins was correlatedto fasting Glu(r=0.62,P=0.001),Within the PCOS group VEGF was correlatedto fasting Glu(r=0.65,P= 0.001),Within the PCOS group T was correlated tofasting Ins (r=0.70,P=0.001),Within the PCOS group VEGF was correlatedto fasting Ins(r=0.41,P=0.01),Within the PCOS group IGF-Ⅱwas correlatedto fasting IGF-Ⅰ(r=0.378,P=0.019).⒌ Results by optical microscope andelectron microscope showed that Ir was detected in the cellularmembrane,mesenchymal cells, thecal cells and gralose cells,especiallyabundant in the vascular endothelial cells,and it was not different between thePCOS and control groups,the PCOS of A canthosis nigricans masc and Acanthosis nigricans negative groups and the PCOS of obesity and non-obesitygroups. ⒍I n all of the ovarian,no IGF-Ir immunoreactivity was seen.⒎ Inovary, immunostaining for IGF-Ⅰ were observed in the theca cells and in thegranulose cells of follicle in PCOS and control groups.There was no staining ofIGF-Ⅰ in the mesenchymal cells. Staining for IGF-Ⅰ in the granulose cells inPCOS were more than control groups(P<0.05).Whereas similar in the thecacells between two groups.⒏ In ovary, immunostaining for VEGF wereobserved in the granulose cells, theca cells, mesenchymal cells andfollicular fluid of follicle in PCOS and control groups. Staining for VEGF inthe granulose cells and mesenchymal cells in PCOS were more than controlgroups(P<0.05, P<0.05).Whereas similar in the theca cells between twogroups. Staining for VEGF in the secondary follicular were more than in theprimary follicular in PCOS.⒐ In ovary, In situ hibridization for IGF-1mRNAwere observed in the theca cells and in the granulose cells of follicle in PCOSand control groups.There was no staining of IGF-1mRNA in the mesenchymalcells. Staining for IGF-1mRNA in the granulose cells in PCOS were more thancontrol groups(P<0.05).Whereas similar in the theca cells between twogroups.⒑ In ovary, In situ hibridization for VEGFmRNA were observed in thegranulose cells, theca cells, mesenchymal cells,cell matrix and follicularfluid of follicle in PCOS and control groups. Staining for VEGFmRNA in thegranulose cells and mesenchymal cells in PCOS were more than controlgroup(P<0.05, P<0.05).Whereas similar in the theca cells between twogroups. Staining for VEGFmRNA in the secondary follicular were more than inthe primary follicular in PCOS.Conclusion: 1. Observing the sufferers of PCOS and their clinical datasrelated to metabolism of glucose and lipid manifests the PCOS' sufferers existthe serious metabolic disorder of glucose and lipid, the ovary blood vesselincreases to live unusually, the mesenchyma in PCOS were more than incontrol groups.2. The serum IGF-Ⅰ,Ins, T level were significantly higher thanthose of the control groups, the serum T level and empty the stomach serum Inspresent obvious straight line relativity, empty the stomach serum Ins withempty the stomach serum Glu present obvious straightly relativity. In sufferer'sbody of PCOS the increased IGF-I did not regulate the function of themetabolism of glucose and lipid same to Ins'by competing to combine the Ⅰr,then participated the formation of insulin resistance of the PCOS sufferers.Serious IR, serious high insulin blood disease may cause A canthosis nigricans,The IR, CHI and high T may exist the mutual promotive infernal circle, it causesthe glucose and lipid metabolic disorder that is not neglected,.All kinds oftherapies of sensitization of insulin are significant to plognosis,we should focuson the T therapy. 3. In PCOS IGF-Ⅱin follicular fluid level to present with theempty stomach serum IGF-Ⅰ level obviously and straightly related, IGF-Ⅱinfollicular fluid level is low,the repression to the output function of the malehormone dies down.Whereas the serum Ins, level of IGF-Ⅰ crease, throughvarious paths promote the male hormone synthesize and secrete, it cause itsfollicular growth obstacle, the choice of the advantage follicular is blocked , agreat deal of small antrum follicular accumulates and ovulate unusually ,thesehave the important function in PCOS occurrence and development.4. Each cellcomposition of the ovary organizes including the gralose cells,thecal cells andstromal cells all have the expression of Ir. The ovary is subjected to the bodyexpression to all regard Ir as principle, and did not discover the expression ofIGF-Ir, the main concentration of Ir is express in afferent surroundings, theothelialcells expression is abundant in the blood vessel particularly, at increasethe expression of living the blood vessel more obviously, it can calculate theovary is another organ of the function of Ins.The Ins and IGFs mainly pass tocombine with Ir but produce result and may have the incitement the afferent andborn function also in the ovary, this function may be one of the paths that Insand IGFs promote the PCOS development. Expression of the Ins and Irs mayparticipate the regulation of ovary function , participate the occurrence and thedevelopment of the PCOS. Ir mainly express surrounding of vessels, thisbrought the inspiration to clinic treatment of the PCOS:When use ovaryfollicular drip to treat the PCOS by the belly cavity mirror , drip ovary surfaceblood vessel to increase to living the area possibly, with the biggest possibilityof bad be subjected to the body, stop the Ins function path, may attain the bettertreatment result.( decrease the male hormone level, stop the male hormone andthe vicious cycle of the insulins).5. At the PCOS ovary gralose cells IGF-1protein and the IGF-1mRNA express high, they combine with the gralose cellsof a great deal ofⅠ r, the IGF-Ⅰ r , make the follicular atresia, stop growth.It isthe important function in PCOS occurrence and development.These have notbeen repoted yet in china.6. The level of serum VEGF and the follicular fluidVEGF is significantly higher than it of normal control groups, the serum VEGFlevel with empty the stomach serum Ins present obviously and straightly related,the VEGF protein and VEGFmRNA expression of primary and the secondaryfollicular step obviously from weak to strong, the secondary follicular expresshigh, the gralose cells and mesenchymal cells VEGF of the ovary expressesstrongly.The insulin may increase the gene of VEGF to transcribe ,then increasethe level of VEGF , the VEGF can urge to increase a quality of ovary bloodvessel to turn, the follicular surroundings blood vessel is born, participating thePCOS occurrence and development. The expression of VEGF in the PCOSovary have not been repoted yet in china.