Effects of constituents from Schisandra chinensis (Wuweizi) on six liver microsomal CYP450 isozymes (CYP1A2, CYP2C6, CYP2C11, CYP2D2, CYP2E1 and CYP3A1/2) were studied in rats in vivo and in vitro. The in vitro incubation was conducted using liver microsomes of rats after multiple dosing of alcoholic/water extract from Schisandra chinensis. A HPLC-MS method was applied to determine the metabolites formation of six CYP450s probe substrates (phenacetin-CYP1A2, dextromethorphan-CYP2D2, diclofenac sodium-CYP2C6, mephenytoin-CYP2C11, chlorzoxazone-CYP2E1 and midazolam-CYP3A1/2) in rat liver microsomal incubations. The activity of CYP450 isozymes were represented by the formation of metabolites. Alcoholic extract of Schisandra chinensis (28-120 microg x mL(-1)) showed significant inhibitory effect on six CYP450 isozymes to a certain extent in vitro. Multiple dosing of Schisandra chinensis alcoholic extract (1.5 g x kg(-1), qd x 7d) had significant induction on CYP2E1 and CYP3A1/2, inhibition on CYP2D2 and CYP2C11, and no effect on CYP2C6 and CYP1A2. Water extract of Schisandra chinensis (100-500 microg x mL(-1)) also exhibited inhibition on the activity of CYP450 isozymes in vitro, whereas multiple administrations (1.5 g x kg(-1), qd x 7d) had significant induction of CYP2E1 and inhibition on CYP2D2, no effect on CYP2C6, CYP3A1/2, CYP1A2 or CYP2C11. The results suggested that the constituents from Schisandra chinensis exhibited the inhibition and induction on six rat liver microsomal CYP450 isozymes to a certain extent in vivo and in vitro. The possibility of interaction between Schisandra chinensis and coadministrative drugs will be considered base on the levels and subtype of CYP450 involved in the drug metabolism.
Animals ; Aryl Hydrocarbon Hydroxylases ; metabolism ; Cytochrome P-450 CYP1A2 ; metabolism ; Cytochrome P-450 CYP2E1 ; metabolism ; Cytochrome P-450 CYP3A ; metabolism ; Cytochrome P-450 Enzyme System ; metabolism ; Cytochrome P450 Family 2 ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Isoenzymes ; metabolism ; Lignans ; isolation & purification ; pharmacology ; Microsomes, Liver ; enzymology ; Plants, Medicinal ; chemistry ; Rats ; Rats, Sprague-Dawley ; Schisandra ; chemistry ; Steroid 16-alpha-Hydroxylase ; metabolism ; Steroid 21-Hydroxylase ; metabolism ; Substrate Specificity

The present study was performed to investigate the effect of bicyclol on hepatic microsomal cytochrome P450 (CYP) activity, as well as gene and protein expressions in rats after partial hepatectomy (PH). Bicyclol (300 mg x kg(-1)) was given to rats subjected to 70% hepatectomy three times before operation. At 6 and 48 h after PH, blood and liver tissue samples were collected for the measurement of serum alanine aminotransferase (ALT), hepatic microsomal malondialdehyde (MDA) and total hepatic CYP content. The activities of four CYP isozymes were detected with liquid chromatography-mass spectrometry (LC-MS) and the gene and protein expressions were determined by RT-PCR and Western blotting assay. As a result, bicyclol pretreatment markedly inhibited the elevation of serum ALT and hepatic microsomal MDA, and prevented the decrease of total hepatic CYP content in PH rats. In addition, bicyclol significantly attenuated the reduction of CYP2C6 activity and mRNA expression, as well as the reduction of CYP2C11 activity in PH rats. Bicyclol can inhibit the decrease of CYP3A1/2 activity, and up-regulate the mRNA and protein expressions of CYP3A1 and CYP2E1. These results showed that bicyclol pretreatment might ameliorate abnormality in CYP450 isoforms during liver regeneration after PH, and this protective effect was likely due to its anti-oxidative property and enzyme induction.
Alanine Transaminase ; blood ; Animals ; Antioxidants ; pharmacology ; Aryl Hydrocarbon Hydroxylases ; genetics ; metabolism ; Biphenyl Compounds ; pharmacology ; Cytochrome P-450 CYP2E1 ; genetics ; metabolism ; Cytochrome P-450 CYP3A ; genetics ; metabolism ; Cytochrome P-450 Enzyme System ; metabolism ; Cytochrome P450 Family 2 ; Enzyme Activation ; drug effects ; Enzyme Induction ; drug effects ; Hepatectomy ; Male ; Malondialdehyde ; metabolism ; Membrane Proteins ; genetics ; metabolism ; Microsomes, Liver ; metabolism ; RNA, Messenger ; metabolism ; Rats ; Rats, Sprague-Dawley ; Steroid 16-alpha-Hydroxylase ; genetics ; metabolism ; Steroid 21-Hydroxylase ; genetics ; metabolism

CYP21A2 gene mutations in a child with congenital adrenal hyperplasia (CAH), and the child's parents, were detected in the study. The clinical features, treatment monitoring and molecular genetic mechanism of CAH are reviewed. In the study, DNA was extracted from peripheral blood samples using the QIAGEN Blood DNA Mini Kit; a highly specific PCR primer for CYP21A2 gene was designed according to the sequence difference between CYP2lA2 gene and its pseudogene; the whole CYP2lA2 gene was amplified with PrimeSTAR DNA polymerase (Takara), and the amplification product was directly sequenced to detect and analyze CYP2lA2 gene mutation. The child was clinically diagnosed with CAH (21-hydroxylase deficiency, 21-OHD) at the age of 36 days, and the case was confirmed by genetic diagnosis at the age of 1.5 years. The proband had a homozygous mutation at c.293-13C in the second intron of CYP21 gene, while the parents had heterozygous mutations. Early diagnosis and standard treatment of CAH (21-OHD) should be performed to prevent salt-wasting crisis and reduce mortality; bone aging should be avoided to increase final adult height (FAH), and reproductive dysfunction due to oligospermia in adulthood should be avoided. These factors are helpful for improving prognosis and increasing FAH. Investigating the molecular genetic mechanism of CAH can improve recognition and optimize diagnosis of this disease. In addition, carrier diagnosis and genetic counseling for the proband family are of great significance.
17-alpha-Hydroxyprogesterone ; blood ; Adrenal Hyperplasia, Congenital ; blood ; genetics ; Humans ; Infant ; Male ; Mutation ; Steroid 21-Hydroxylase ; genetics

OBJECTIVE: Some, but not all, granulosa cell tumors are characterized by estrogen production. This study was designed to determine whether there are clinical or pathological variations in granulosa cell tumors in relation to the expression of sex steroid synthesis enzymes. METHODS: Clinical symptoms, serum hormonal values, and histology of 30 granulosa cell tumor patients who underwent surgery between 2002 and 2014 were retrospectively reviewed. RESULTS: Most patients presented with abnormal genital bleeding including abnormal menstrual cycles. Eight of 16 patients older than 50 years had endometrial hyperplasia and one had endometrial cancer. Serum 17β-estradiol (E2) levels tended to be higher in patients over 50 years of age (p=0.081). Serum follicle-stimulating hormone (FSH) levels were low in all patients irrespective of serum E2 levels. Magnetic resonance imaging revealed a thicker endometrium in older as compared to younger patients (p<0.05). Tumor cells in the majority of cases were positive for inhibin α and P450 aromatase, irrespective of age and serum E2 levels. P450 17α-hydroxylase (P450c17) expression varied among cases. P450c17 was strongly positive in luteinized tumor cells and weakly positive in theca cells and fibroblasts. High E2 levels were associated with P450c17-positive cells in the tumor (p<0.05). CONCLUSION: The expression of hormone-synthesizing enzymes divides granulosa cell tumors into 2 distinct types; tumors with P450c17-positive cells show elevated serum E2 and related clinical symptoms, while tumors without these cells show symptoms related to FSH suppression by inhibin.
Adult* ; Aromatase ; Endometrial Hyperplasia ; Endometrial Neoplasms ; Endometrium ; Estrogens ; Female ; Fibroblasts ; Follicle Stimulating Hormone ; Granulosa Cell Tumor* ; Granulosa Cells* ; Hemorrhage ; Humans ; Inhibins ; Lutein ; Magnetic Resonance Imaging ; Menstrual Cycle ; Ovary* ; Retrospective Studies* ; Steroid Hydroxylases ; Theca Cells

A single measurement of serum 17alpha-hydroxyprogesterone (17OHP) level can be unreliable because of its marked diurnal variation. We investigated the relationship of serum level of 17OHP with that of androstenedione (AD), which shows a smaller diurnal variation. And we tested whether the responses of these two hormones to low-dose ACTH stimulation are correlated in patients with 21-hydroxylase deficiency. Baseline serum 17OHP and AD levels were measured in 87 patients and a low-dose ACTH stimulation test was performed in 41 patients. The basal 17OHP level correlated positively with the basal AD level independently of sex, type of 21-hydroxylase deficiency, and the time of day of blood sampling (n = 87, R2 = 0.75, P < 0.001). The area under the curve of 17OHP and AD correlated positively with their respective basal levels. The fold-change increase in 17OHP after ACTH injection correlated negatively with the basal 17OHP level, but that of AD did not correlate with the basal AD level. The random serum 17OHP level, used in the clinic, is a reliable guide and a low-dose ACTH stimulation test provides no extra benefit for assessing the treatment adequacy in patients with 21-hydroxylase deficiency.
17-alpha-Hydroxyprogesterone/*blood ; Adolescent ; Adrenal Hyperplasia, Congenital/*diagnosis/drug therapy ; Adrenocorticotropic Hormone/*diagnostic use ; Androstenedione/*blood ; Circadian Rhythm ; Female ; Humans ; Male ; Steroid 21-Hydroxylase/metabolism ; Young Adult

OBJECTIVE: We analyzed the degree of behavioral and psychic masculinization as well as factors influencing the masculinization in female patients with CAH caused by 21-hydroxylase deficiency. METHODS: We studied 26 patients with salt-wasting (SW) type, 29 patients with simple virilizing (SV) type and 30 control patients, who were older than 10 years of age. Study participants were given a questionnaire containing 55 items (20 masculine items, 20 feminine items, 15 neutral items) in order to evaluate the degree of feminity and masculinity. RESULTS: SW and SV girls revealed significantly higher masculine scores and lower feminine scores than the control groups (P<0.05). Feminine scores were lower in the SW group compared to those in the SV group (P<0.05). Neutral scores showed no significant difference among three groups. Masculine, feminine, bisexual and undifferentiated personality characteristics exhibit 57.6%, 3.8%, 7.6%, and 30.7% in SW group, respectively; 31%, 17.2%, 20.6%, and 30%, respectively in SV group; 3.3%, 50.0%, 20.0% and 26.6%, respectively in control group. Masculine scores were negatively correlated with age at diagnosis and treatment (r=-0.516, P<0.05), but not with the levels of 17-hydroxyprogesterone at the time of diagnosis and study, advancement of bone age, age at clitoplasty, age at onset of puberty and age at menarche. CONCLUSIONS: Female patients with 21-hydroxylase deficiency, who reveal high masculinizing score, should be closely monitored for the adverse effects of high degree of masculinization, such as polycystic ovarian syndrome. Additionally, it is important to treat adequately after early diagnosis to minimize the degree of masculinization and to develop the prenatal mode of treatment to interrupt the secretion of adrenal androgen.
17-alpha-Hydroxyprogesterone ; Adolescent ; Adrenal Hyperplasia, Congenital* ; Bisexuality ; Diagnosis ; Early Diagnosis ; Female* ; Humans ; Masculinity ; Menarche ; Polycystic Ovary Syndrome ; Puberty ; Surveys and Questionnaires ; Steroid 21-Hydroxylase*

BACKGROUND: High levels of 17-hydroxyprogesterone (17-OHP) are frequently observed in premature infants without congenital adrenal hyperplasia. The purpose of this study is to set cut-off limits of 17-OHP on the basis of gestational age at birth and birth weight. METHODS: Blood spot 17-OHP concentrations were measured in 1,000 infants on the 3th day of life at Inha University Hospital. An enzyme-linked immunosorbent assay (ELISA) method (ICN Neoscreen ELISA 17-hydroxyprogesterone kit, ICN Pharmaceuticals. Inc., Japan) was used. The values obtained were analyzed with respect to birth weight and gestational age at birth in order to decide the appropriate cut-off limits in a neonatal mass screening for 21-hydroxylase deficiency. RESULTS: In the neonatal mass screening for CAH, the cut-off limits for determining the 17-OHP for recall, were decided as follows: (1) 57.65, 39.88, 33.52 ng/mL for gestational age at birth of 35 weeks or less, 36-37, and 38 weeks or more, respectively, and (2) 54.88, 43.86, 32.92 ng/mL for birth weight of 2.49 or less, 2.50-2.99, 3.00 kg or more, respectively. CONCLUSIONS: The cut-off limits on the basis of gestational age at birth and birth weight should be used in the screening for congenital adrenal hyperplasia. We believe that the false positive rate in premature infants can be reduced using this method.
17-alpha-Hydroxyprogesterone* ; Adrenal Hyperplasia, Congenital ; Birth Weight* ; Enzyme-Linked Immunosorbent Assay ; Gestational Age* ; Humans ; Infant ; Infant, Newborn ; Infant, Premature ; Mass Screening ; Neonatal Screening ; Parturition* ; Steroid 21-Hydroxylase

We experienced a boy manifesting sexual precocity with unilateral testicular tumor, who was finally diagnosed as CAH with 21-hydroxylase deficiency. Initial laboratory findings were compatible with peripheral precocious puberty. Ultrasonogram for testes showed heterogenous high echoic mass in the right testicle. Radical orchiectomy was performed and its pathology revealed benign Leydig cell tumor without Reinke crystal. But, testosterone was not decreased after 1 month of surgery. Second laboratory exam revealed increased DHEA-S and 17-hydroxyprogesterone. Finally, this case was recognized as CAH(simple virilizing type) with unilateral testicular adrenal rest tumor. In conclusion, CAH should always be considered during etiologic study for the male sexulal precocity even with unilateral testicular tumor.
17-alpha-Hydroxyprogesterone ; Adrenal Hyperplasia, Congenital ; Adrenal Rest Tumor ; Humans ; Leydig Cell Tumor* ; Male ; Orchiectomy ; Pathology ; Puberty, Precocious ; Steroid 21-Hydroxylase ; Testis ; Testosterone ; Ultrasonography

Congenital adrenal hyperplasia is inherited disorder of adrenal steroidogenesis. 21-hydroxylase deficiency is the most commone enzymatic defect and is divided into classic and late-onset or nonclassic forms. Both classic non-classic 21-hydrozylase deficiencies are inherited in a recessive manner as allelic variants. But it is rare that happened in twin infants. Chief complaints of affected twins in our case were ambiguous genitalia, hyperpigmentation and dehydrations. They were revealed into hyponatremia, hyperkalemia and increased amount of serum progesterone, 17-hydroxyprogesterone and urinary 17-ketosteroid excretion and were administered with DOCA, 9alpha-fluorohydrocortisone, hydrocortisone to control the electrolyte imbalance. And now, both of them are going to normal ratio of weight gain and body growth.
17-alpha-Hydroxyprogesterone ; Adrenal Hyperplasia, Congenital* ; Desoxycorticosterone Acetate ; Disorders of Sex Development ; Humans ; Hydrocortisone ; Hyperkalemia ; Hyperpigmentation ; Hyponatremia ; Infant ; Progesterone ; Steroid 21-Hydroxylase* ; Twins* ; Weight Gain

The fields of pharmacogenetics and pharmacogenomics have become increasingly promising regarding the clinical application of genetic data to aid in prevention of adverse reactions. Specific screening tests can predict which animals express modified proteins or genetic sequences responsible for adverse effects associated with a drug. Among the genetic variations that have been investigated in dogs, the multidrug resistance gene (MDR) is the best studied. However, other genes such as CYP1A2 and CYP2B11 control the protein syntheses involved in the metabolism of many drugs. In the present study, the MDR-1, CYP1A2 and CYP2B11 genes were examined to identify SNP polymorphisms associated with these genes in the following four canine breeds: Uruguayan Cimarron, Border Collie, Labrador Retriever and German Shepherd. The results revealed that several SNPs of the CYP1A2 and CYP2B11 genes are potential targets for drug sensitivity investigations.
Animals ; Aryl Hydrocarbon Hydroxylases/*genetics/metabolism ; Cytochrome P-450 CYP1A2/*genetics/metabolism ; Dogs/*genetics/metabolism ; P-Glycoprotein/*genetics/metabolism ; *Polymorphism, Single Nucleotide ; Steroid Hydroxylases/*genetics/metabolism