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《JOURNAL OF REPRODUCTIVE MEDICINE》 1995-01
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The steroid 5a-reductase isozyme activities in foreskin of Chinese adult men

CUI Yugui,ZHANG Guiyuan Department of Reproductive Endocrinology,National Research Institute for Family Planning,Beijing 100081  
bjectives :To develop a method for measuring the steroid 5a- eductase isozyme type I and type Il ac-tivities in foreskn of Chinese adult men.Design:A laboratorgy methodology and its clinical application.setting:The endocrinology laboratory of National Research Institute for Family Planning.Subjects: 22 Chinese adult men. Intervention :Foreskin specimens were taken from the patients at the time of circumcision with consentand were quickly frozen in liquid nitrogen and stored at一7O℃. Methods :The procedure of the method includes:homogenization of tissue in Tris/chloride buffer on ice;centrifugation of supernatant at 120 000g to get microsomal peIlets and resuspended by Dounce homogeniza-tion :standard assay of 5a- reductase isozyme activities of microsomal pellet;labelled testosterone used as sub-strate; steroids were extracted and separated with thin layer chromatography;3 H-dihydrotestosteroneformed was counted for radioactivitv. Results :The steroid 5a- reductase isozyme activities were characterized by pH optima 5.5 and a broaderrange of 7~9,and exhibit different kinetics with respect to substrate testosterone. Km values for theisozvme II and I were 0.079 and 0.46μmol/L,respectively.The isozyme I and II activities in foreskin of22 Chinese adult men were 21.15± 1 53 and 50.8335.70(x 士s) pmoles DHT formation/mg protein/30min,respectively. Conclusions :This method of measuring 5a-reductase activities has the advantages of being stable,highlysensitive,easy to perform and more accurate than RIA of DH It can be used in clinical practice for diagno-sis of 5a-reductase deficiencv and other sexual differentiation disorders.It is also valuable in the basic re- search on BPH and hurman reproduction.
【CateGory Index】: R339.2
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