| It has been confirmed that proliferation of tunica intima, proliferation and hypertrophy of smooth muscle in tunica mediain in the early and middle stage,apoptosis of smooth muscle and shedding of arteriosclerotic plaque are the characteristic of arteriosclerosis.The cell and tissue of artery experienced a pathologic process that increasing at first and then decreasing at last.But the underlying mechanism remains unknown. Previous studies on the mechamism of arteriosclerosis pay more attention to the changes of growth factor and extracellular matrix. Recent studies indicated that Heme oxygenase is a restrictive enzyme which catalyzes oxidizing degradation of protoheme in vivo;On the spot there are three categories of isoenzymes detected HO-1 , HO-2 , HO-3 ; Protoheme catalyzed by HO could generate CO bilirubin and sider-ion.HO-1 is a inducible isoform which have antagonism against oxidative stress injury. The means of HO-1 induction by hypoxia in arteriosclerosis was to generating of CO,the latter could inhibit proliferation , hypertrophy and apoptosis of smooth muscle through many pathway .But it remains unknown that l)if HO-1 in artery would increased in dibetic arteriosclerosis.2)the main pathway that CO suppress proliferation,hypert-rophy and apoptosis of smooth muscle. Previous studies focused on the heart blood vessal, the brain blood vessal and the pulmonary blood vessal to study HO qualitation, location and quantitation. The interralated reports on relation of HO and the testis artery have been found. In this study KKAy dibetic mice aged 5w~21w were used to investigate the dynamic changes of proliferation,hypertrophy and apoptosis of smooth muscle and the possible mechanism in arteriosclerosis.The natural episodic diabete model was used to observe proliferative and hypertrophic changes in smooth muscle of testicular arteries .The correlative genetic changes was investigated by gene chip technique.The experimental animals were divided into two groups:KKAy diabetic (diabetes mellitus,DM)group and control group (C57Blmice). Diabetic group was then divided into 7 groups,which were 5w?7w, 9w, llw, 16w, 21 w, 30w aged KKAy diabetic mice groups,each had 6 animals.Control groups were age-matched male C57BL mice. BrdU incorporation ,cellular proliferating cycle and ultrastructure were studied using histochemistry, immunohistochemistry, transmission electron microscope, vascular cast of scanning electronmicroscope and flow cytometry.Total RNA was abstracted with one-step method and mRNA was purified with Oligo-dt cellulose,cDNA probes labeled with cy3 and cy5 were obtained by reverse transcription-polymerase chain reaction(RT-PCR).Hybridization of cDNA probe and gene chip was carried out,gene chip was scanned with Scan Array3000,and results were analyzed with ImaGene 3.0.Result: 1 Toluidine blue staining of semithin section showed: compared with control group,the boundary of tunica intima,tunica media and tunica adventitia was not distinct, tunica intima and internal elastic membrane was thickened and multilaminar.2 Masson staining:In DM groups,smooth muscle in tunica media and connective tissue in tunica adventitia remarkably increased,especially in 9w llw and 16w group.3 Immunohistochemistry and flow cytometry of cell proliferation :3.1 BrdU corporation of testicular artery: Few positive cells were observed during the whole procedure in control group while positive cells spreaded in three layer of artery, especially in tunica media. The number of positive cells was obviously increased in 9w group, peaked in llw group and then decreased in 16w group.3.2 Flow cytometry: S phase G2 phase and M phase are proliferative phases.The proliferation in control group was in low level while proliferative cells gradually increased in 5w 7w 9w llw DM groups , peaked in llw,decreased in 16w and reached its minimum in 3 Ow. Significant difference existed between 9w llw and 21w 30w group. 4 infrastructure:4.1 The abnormal ultrastructure was rarely seen in 5w testicular artery. TEM showed the intima of testicu...
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