Carbon Chain Length Determines Inhibitory Potency of Perfluoroalkyl Sulfonic Acids on Human Placental 3β-Hydroxysteroid Dehydrogenase 1:Screening,Structure-Activity Relationship,and In Silico Analysis

Objective This study aimed to compare 9 perfluoroalkyl sulfonic acids(PFSA)with carbon chain lengths(C4–C12)to inhibit human placental 3β-hydroxysteroid dehydrogenase 1(3β-HSD1),aromatase,and rat 3β-HSD4 activities.Methods Human and rat placental 3β-HSDs activities were determined by converting pregnenolone to progesterone and progesterone secretion in JEG-3 cells was determined using HPLC/MS–MS,and human aromatase activity was determined by radioimmunoassay.Results PFSA inhibited human 3β-HSD1 structure-dependently in the order:perfluorooctanesulfonic acid(PFOS,half-maximum inhibitory concentration,IC50:9.03±4.83μmol/L)>perfluorodecanesulfonic acid(PFDS,42.52±8.99μmol/L)>perfluoroheptanesulfonic acid(PFHpS,112.6±29.39μmol/L)>perfluorobutanesulfonic acid(PFBS)=perfluoropentanesulfonic acid(PFPS)=perfluorohexanesulfonic acid(PFHxS)=perfluorododecanesulfonic acid(PFDoS)(ineffective at 100μmol/L).6:2FTS(1H,1H,2H,2H-perfluorooctanesulfonic acid)and 8:2FTS(1H,1H,2H,2H-perfluorodecanesulfonic acid)did not inhibit human 3β-HSD1.PFOS and PFHpS are mixed inhibitors,whereas PFDS is a competitive inhibitor.Moreover,1–10μmol/L PFOS and PFDS significantly reduced progesterone biosynthesis in JEG-3 cells.Docking analysis revealed that PFSA binds to the steroid-binding site of human 3β-HSD1 in a carbon chain length-dependent manner.All 100μmol/L PFSA solutions did not affect rat 3β-HSD4 and human placental aromatase activity.Conclusion Carbon chain length determines inhibitory potency of PFSA on human placental 3β-HSD1 in a V-shaped transition at PFOS(C8),with inhibitory potency of PFOS>PFDS>PFHpS>PFBS=PFPS=PFHxS=PFDoS=6:2FTS=8:2FTS.

Effects of genistein and equol on human and rat testicular 3β-hydroxysteroid dehydrogenase and 17β-hydroxysteroid dehydrogenase 3 activities

The objective of the present study was to investigate the effects of genistein and equol on 3β-hydroxysteroid de- hydrogenase （3β-HSD） and 17β-hydroxysteroid dehydrogenase 3 （17β-HSD3） in human and rat testis microsomes. These enzymes （3β-HSD and 17β-HSD3）, along with two others （cytochrome P450 side-chain cleavage enzyme and cytochrome P450 17α-hydroxylase/17-20 lyase）, catalyze the reactions that convert the steroid cholesterol into the sex hormone testosterone. Genistein inhibited 3β-HSD activity （0.2 μmol L^-1 pregnenolone） with half-maximal inhibition or a half-maximal inhibitory concentration （IC50） of 87 ± 15 （human） and 636 ± 155 nmol L^-1 （rat）. Genistein＇s mode of action on 3β-HSD activity was competitive for the substrate pregnenolonrge and noncompetitive for the cofactor NAD＋. There was no difference in genistein＇s potency of 3β-HSD inhibition between intact rat Leydig cells and testis microsomes. In contrast to its potent inhibition of 3β-HSD, genistein had lesser effects on human and rat 17β-HSD3 （0.1 μmol L^-1 androstenedione）, with an IC50 〉 100μmol L^-1. On the other hand, equol only inhibited human 3β-HSD by 42%, and had no effect on 3β-HSD and 17β-HSD3 in rat tissues. These observations imply that the ability of soy isoflavones to regulate androgen biosynthesis in Leydig cells is due in part to action on Leydig cell 3β- HSD activity. Given the increasing intake of soy-based food products and their potential effect on blood androgen levels, these findings are greatly relevant to public health.

11β-hydroxysteroid dehydrogenase types 1 and 2. in postnatal development of rat testis： gene express,on, localization and regulation by luteinizing hormone and androgens

11β-hydroxysteroid dehydrogenase type 1 （11β-HSD1） and type 2 （11β-HSD2） are expressed in rat testis, where they regulate the local concentrations of glucocorticoids. Here, we investigated the expression and localization of 11β-HSD in rat testis during postnatal development, and the regulation of these genes by luteinizing hormone （LH） and androgens, mRNA and protein levels were analyzed by quantitative real-time-polymerase chain reaction and western blotting, respectively, in testes collected from rats at postnatal day （PND） 7, 14, 21, 35, and 90, and from rats treated with LH, 7α.methyl-19-nortestosterone （MENT） and testosterone at PND 21 and PND 90. Immunohistochemical staining was used to identify the localization of the 11β-HSD in rat testis at PND 7, 14, and 90. We found that 11β-HSD1 expression was restricted to the interstitial areas, and that its levels increased during rat testis development. In contrast, whereas 11β-HSD2 was expressed in both the interstitial areas and seminiferous tubules at PND 7, it was present only in the interstitial areas at PND 90, and its levels declined during testicular development. Moreover, 11β-HSD1 mRNA was induced by LH in both the PND 21 and 90 testes and by MENT at PND 21, whereas 11β-HSD2 mRNA was induced by testosterone and MENT in the PND 21 testis and by LH in the PND 90 testis. In conclusion, our study indicates that the 11β-HSD1 and 11β-HSD2 genes have distinct patterns of spatiotemporal expression and hormonal regulation during postnatal development of the rat testis.

Increased Expression of 11<i>β</i>-Hydroxysteroid Dehydrogenase Type 1 in Experimental Periodontitis Induced by Lipopolysaccharide from <i>Porphyromonas gingivalis</i>

It has been proposed that 11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1), which activates glucocorticoids, plays a role in chronic inflammatory diseases including metabolic diseases, rheumatoid arthritis, and ulcerative colitis. We have recently reported that the expression of 11β-HSD1 is increased in the gingiva of patients with chronic periodontitis and in that of rats with ligature-induced periodontitis. In this study, to further demonstrate the involvement of 11β-HSD1 in chronic periodontitis, the expression of 11β-HSD1 was investigated in another rat model of experimental periodontitis induced by intragingival injection of lipopolysaccharide from Porphyromonas gingivalis (LPS-PG). Alveolar bone loss was observed two weeks after intragingival injection of LPS-PG. The level of 11β-HSD1 mRNA assessed by real-time reverse transcriptase-polymerase chain reaction was significantly elevated in LPS-PG-induced periodontitis compared with controls. The expression of 11β-hydroxysteroid dehydrogenase type 2 (11β-HSD2), which inactivates glucocorticoids, was not significantly different between control and LPS-PG-induced periodontitis. The expression of 11β-HSD1 was significantly correlated with that of TNF in LPS-PG-induced periodontitis. The increased expression of 11β-HSD1 protein in LPS-PG-induced periodontitis was confirmed by immunohistochemistry using anti-11β-HSD1 antibody. These results further suggest a role for 11β-HSD1 in the pathogenesis of chronic periodontitis.

大鼠肾脏细胞17β-HSD1的表达及参与性激素合成的能力

目的通过研究合成性激素的关键酶17β-HSD1在肾脏中的表达,探讨肾脏是否具备合成性激素的作用。方法基于无促卵泡生成素(FSH)、黄体生成素(LH)培养基和有FSH、LH培养基两种条件下,Western blotting、放射免疫分析法分别检测培养24、48 h后肾脏细胞中17β-HSD1的表达和性激素的分泌情况。结果培养24 h后,大鼠肾脏细胞能够表达少量的17β-HSD1蛋白(0.1843±0.076),同时能够分泌少量的雌二醇、孕酮和睾酮(分别为3.30±3.78 nmol/L,62.60±12.33 pmol/L和22.12±3.36 nmol/L),而在FSH和LH的共同刺激下,大鼠肾脏细胞17β-HSD1蛋白的表达量明显升高(1.6651±0.044,P<0.01),同时分泌雌二醇、孕酮和睾酮的量也显著增加(分别为8.50±2.64 nmol/L,117.80±9.79 pmol/L和45.04±4.39 nmol/L,均P<0.05),培养24h和48 h上述指标均无明显差异(P>0.05)。结论大鼠肾脏细胞中有17β-HSD1的表达,并且在FSH和LH的共刺激下能够稳定分泌性激素,提示肾脏组织具备合成性激素的能力,丰富了肾脏的内分泌功能。

雌激素受体和17β-羟类固醇脱氢酶1在子宫内膜息肉中的表达及其意义

目的研究雌激素受体(estrogen receptor,ER)和17β-羟类固醇脱氢酶1(17β-HSD1)在子宫内膜息肉(endometrial polyps,EPs)中的表达情况,进一步从分子水平阐明EPs的发病机制。方法选择EPs患者47例,其中,息肉组织作为病例组,同一患者宫腔内远离息肉组织的正常内膜作为对照1组,同时选择50例行宫腔镜检查的妇女的正常子宫内膜组织作为对照2组。采用免疫组织化学法检测ER和17β-HSD1在各组的表达情况并进行比较。结果 ER在EPs中表达的阳性率是80.9%,17β-HSD1在EPs中表达的阳性率是76.6%,二者在EPs中表达均高于对照组,差异有统计学意义(P<0.01)。ER和17β-HSD1在EPs中的表达呈正相关(r=0.619,P<0.01)。结论 ER和17β-HSD1在EPs中的过度表达可能是EPs发生、发展的原因。

环腺苷一磷酸对人Ⅰ型17β羟类固醇脱氢酶在绒癌细胞系中表达的调节作用

由HSD17B1基因编码的人Ⅰ型17β-羟类固醇脱氢酶(17β-hydroxysteroid dehydrogenasetype 1,简称Ⅰ型17HSD)催化雌酮与雌二醇之间的转化。本文研究环腺苷一磷酸简称(cAM-P)对该酶在培养的绒癌细胞系(JAR和JEG-3)中表达的调节作用。用8-bromo-cAMP处理两种绒癌细胞后,观察到在伴随1.3 kbⅠ型17 HSDmRNA表达的同时,Ⅰ型17 HSD蛋白浓度也显著上升。标记基因分析表明,cAMP可诱导HSD 17 B1基因启动子在JAR和JEG-3细胞系中的转录活性,参与调节这一诱导作用的区域位于HSD 17 B1基因编码区上游-659至-550处。凝胶阻滞实验显示这一区域可同JAR、JEG-3、T-47 D和HeLa细胞核抽提物形成特异的DNA-蛋白复合物。本结果首次证实cAMP激活HSD 17 B1基因启动子在绒癌细胞中的转录。

Correlation between single nucleotide polymorphisms of 17β-HSD-1 and endometrial adenocarcinoma

Objective: To investigate the correlation between 17-beta-hydroxysteroid dehydrogenase type1(17β-HSD-1) gene polymorphisms and risk of endometrial adenocarcino-ma.Methods: Forty-one patients with endometrial adenocarcinoma were selected as experimen-tal group and twenty-seven healthy women were selected as control group.The three common single nucleotide polymorphism of 17β-HSD-1 gene at sites + 1004,+ 1322 and + 1954 were detected by allele-specific PCR(ASA-PCR).The allele frequencies were analyzed by SPSS13.0 software between endometrial cancer cases and controls.Results: We observed no significant difference in various frequency distribution between experimental group and control group.P1004= 0.994,P1322 = 0.974,and P1954 = 0.981.Conclusion: We found that three common SNPs with the 17β-HSD-1 gene were not associated with endometrial adenocarcinoma.We suggest that more research for 17β-HSD needs to explore.

G6PT-H6PDH-11βHSD1 triad in the liver and its implication in the pathomechanism of the metabolic syndrome

The metabolic syndrome, one of the most common clinical conditions in recent times, represents a combination of cardiometabolic risk determinants, including central obesity, glucose intolerance, insulin resistance, dyslipidemia, non-alcoholic fatty liver disease and hypertension. Prevalence of the metabolic syndrome is rapidly increasing worldwide as a consequence of common overnutrition and consequent obesity. Although a unifying picture of the pathomechanism is still missing, the key role of the pre-receptor glucocorticoid activation has emerged recently. Local glucocorticoid activation is catalyzed by a triad composed of glucose-6-phosphate-transporter, hexose-6-phosphate dehydrogenase and 11β-hydroxysteroid dehydrogenase type 1 in the endoplasmic reticulum. The elements of this system can be found in various cell types, including adipocytes and hepatocytes. While the contribution of glucocorticoid activation in adipose tissue to the pathomechanism of the metabolic syndrome has been well established, the relative importance of the hepatic process is less understood. This review summarizes the available data on the role of the hepatic triad and its role in the metabolic syndrome, by confronting experimental findings with clinical observations.

加味三棱丸抗子宫内膜异位症内膜细胞雌二醇生成机制的研究

目的:探讨加味三棱丸(SLW)抗子宫内膜异位症内膜雌激素生成的作用机制。方法:以SLW含药血清作用于体外培养的内异症在位内膜细胞,RT-PCR检测子宫肌瘤组、给予SLW含药血清前后内异症组类固醇类产生因子-1(steroidgenic factor-1,SF-1)、小鸡卵清蛋白上游启动转录因子(chicken ovalbumin upstream-transcription factor,COUP-TF),17-β-羟甾类脱氢酶1(17-beta-hydroxysteroid dehydrogenase 1,17-β-HSD 1)和17-β-羟甾类脱氢酶2(17-beta-hydroxysteroid dehydrogenase 2,17-β-HSD 2)mRNA的表达。Western blot检测上述各组SF-1和COUP-TF蛋白的表达。结果:子宫肌瘤对照组SF-1,17-β-HSD 1 mRNA及SF-1蛋白的表达明显低于内异症内膜组,而COUP-TF,17-β-HSD 2 mRNA及COUP-TF蛋白的表达明显高于内异症在位内膜组,差异有极显著性(P<0.01)。以内异症在位内膜为对照,给予对照组SLW 5.0,2.5 g.kg-1.d-1大鼠含药血清(体积分数为0.1)作用48 h后,COUP-TF mRNA和蛋白、17-β-HSD 2 mRNA的表达明显高于对照组,而SF-1 mRNA和蛋白、17-β-HSD 1 mRNA的表达明显低于对照组,差异有显著性或极显著性(P<0.05或P<0.01)。SLW 1.25 g.kg-1.d-1组COUP-TF mRNA和蛋白的表达水平明显高于对照组,SF-1蛋白和17-β-HSD 1 mRNA表达明显低于对照组(P<0.01),而SF-1,17-β-HSD 2 mRNA的表达却无明显改变。结论:加味三棱丸可降低子宫内膜异位症在位内膜细胞分泌雌二醇的水平,与其抑制SF-1和17-β-HSD 1,增强COUP-TF和17-β-HSD 2的表达密切相关。

表达3-甾酮-△^(1)-脱氢酶降解植物甾醇合成雄甾-1,4-二烯-3,17-二酮

构建分枝杆菌表达载体pMTac并在分枝杆菌Mycobacterium neoaurum JC-12中加强表达甾醇降解过程中的关键酶3-甾酮-△1-脱氢酶(KSDD)以提高雄甾-1,4-二烯-3,17-二铜(ADD)的产量。将p MF41的启动子pACE替换成tac启动子构建载体pMTac,在分枝杆菌中分别表达报告基因绿色荧光蛋白(GFP)和关键酶KSDD,通过GFP亮度和KSDD酶活验证tac启动子在M.neoaurum JC-12中的效果,并发酵验证加强表达KSDD对产物ADD的影响。荧光显微照片表明两个载体均能在M.neoaurum JC-12表达GFP,但tac启动子的效果比pACE强。酶活测定结果为重组菌M.neoaurum JC-12/pMTac-ksdd破碎细胞上清液中KSDD酶活比原始菌提高了6.53倍,比M.neoaurum JC-12/pMF41-ksdd提高了4.36倍。摇瓶发酵显示重组菌M.neoaurum JC-12/pMTac-ksdd ADD的产量比原始菌提高了22.2%,由4.86 g/L提高到5.94 g/L,而AD的产量由0.92 g/L减少到0.17 g/L,降低了81.5%;与M.neoaurum JC-12/p MF41-ksdd比,ADD产量提高了12.7%,AD降低了71.2%。以20 g/L植物甾醇为底物,5 L发酵罐中重组菌M.neoaurum JC-12/pMTac-ksdd的ADD产量达到10.28 g/L。结果表明,构建的新型表达载体pMTac适用于在M.neoaurum JC-12中加强表达关键酶KSDD,而且在M.neoaurum JC-12中过量表达KSDD有助于ADD产量的提高,为目前报道的发酵法利用新金色分枝杆菌降解植物甾醇合成ADD的最高水平。

Synergistic control of sex hormones by 17β-HSD type 7: a novel target for estrogen-dependent breast cancer

17β-hydroxysteroid dehydrogenase(17β-HSD)type 1 is known as a critical target to block the final step of estrogen production in estrogen-dependent breast cancer.Recent confirmation of the role of dyhydroxytestosterone(DHT)in counteracting estrogeninduced cell growth prompted us to study the reductive 17β-HSD type 7(17β-HSD7),which activates estrone while markedly inactivatingDHT.The role ofDHTin breast cancer cell proliferation isdemonstratedby its independent suppression of cell growthin the presence of a physiological concentration of estradiol(E2).Moreover,an integral analysis of a large number of clinical samples in Oncomine datasets demonstrated the overexpression of 17β-HSD7 in breast carcinoma.Inhibition of 17β-HSD7 in breast cancer cells resulted in a lower level of E2 and a higher level of DHT,successively induced regulation of cyclinD1,p21,Bcl-2,and Bik,consequently arrested cell cycle in the G0/G1 phase,and triggered apoptosis and auto-downregulation feedback of the enzyme.Such inhibition led to significant shrinkage of xenograft tumors with decreased cancer cell density and reduced 17β-HSD7 expression.Decreased plasma E2 and elevated plasma DHT levels were also found.Thus,the dual functional 17β-HSD7 is proposed as a novel target for estrogen-dependent breast cancer by regulating the balance of E2 andDHT.Thisdemonstrates aconceptual advance on the general belief that the major role of this enzyme is in cholesterol metabolism.

Novel and emerging diabetes mellitus drug therapies for the type 2 diabetes patient

Type 2 diabetes mellitus is a metabolic disorder of deranged fat, protein and carbohydrate metabolism resulting in hyperglycemia as a result of insulin resistance and inadequate insulin secretion. Although a wide variety of diabetes therapies is available, yet limited efficacy, adverse effects, cost, contraindications, renal dosage adjustments, inflexible dosing schedules and weight gain significantly limit their use. In addition, many patients in the United States fail to meet the therapeutic HbA1c goal of 【 7% set by the American Diabetes Association. As such new and emerging diabetes therapies with different mechanisms of action hope to address some of these drawbacks to improve the patient with type 2 diabetes. This article reviews new and emerging classes, including the sodium-glucosecotransporter-2 inhibitors, 11β-Hydroxysteroid dehydrogenase type 1 inhibitors, glycogen phosphorylase inhibitors; protein tyrosine phosphatase 1B inhibitors, G Protein-Coupled receptor agonists and glucokinase activators. These emerging diabetes agents hold the promise of providing benefit of glucose lowering, weight reduction, low hypoglycemia risk, improve insulin sensitivity, pancreatic β cell preservation, and oral formulation availability. However, further studies are needed to evaluate their safety profile, cardiovascular effects, and efficacy durability in order to determine their role in type 2 diabetes management.