Hydroxysteroid Dehydrogenase Research Articles

Renal 11[beta]-hydroxysteroid dehydrogenase type 2 is of central importance for 11-oxygenated androgen biosynthesis and is disrupted in chronic kidney disease

Renal 11[beta]-hydroxysteroid dehydrogenase type 2 is of central importance for 11-oxygenated androgen biosynthesis and is disrupted in chronic kidney disease

11[beta]-hydroxysteroid dehydrogenase type 1 inhibition unmasks multiple pathways that may mitigate the adverse effects of prescribed prednisolone

11[beta]-hydroxysteroid dehydrogenase type 1 inhibition unmasks multiple pathways that may mitigate the adverse effects of prescribed prednisolone

2 Another renaissance for bile acid gastrointestinal microbiology

Abstract The field of bile acid microbiology in the gastrointestinal tract is going through a current rebirth after a peak of activity in the late 1970s and early 1980s. This renewed activity is a result of many factors including the discovery near the turn of the century that bile acids are potent signaling molecules, technological advances in next generation sequencing, computation, culturomics, gnotobiology, and metabolomics. The current state of the field with particular emphasis on questions that have remained unanswered for many decades in both bile acid synthesis by the host, and metabolism by the gut microbiota will be addressed. Current knowledge of established enzymatic pathways including bile salt hydrolase, hydroxysteroid dehydrogenases involved in the oxidation and epimerization of bile acid hydroxy groups, the Hylemon-Bjӧrkhem Pathway (HBP) of bile acid C7-dehydroxylation, and the formation of secondary allo-bile acids are described. Aspects of bile acid conjugation and esterification, and evidence for bile acid C3- and C12-dehydroxylation that are less well understood, but potentially critical for our understanding of bile acid metabolism in the human gut will be covered. The physiological consequences of bile acid metabolism for human health, important caveats and cautionary notes on experimental design and interpretation of bile acid metabolism are also explored.

HSD17B13 liquid-liquid phase separation promotes leukocyte adhesion in chronic liver inflammation.

The rs72613567:TA polymorphism in 17-beta hydroxysteroid dehydrogenase 13 (HSD17B13) has been found to reduce the progression from steatosis to nonalcoholic steatohepatitis. In this study, we sought to define the pathogenic role of HSD17B13 in triggering liver inflammation. Here we find that HSD17B13 forms liquid-liquid phase separation (LLPS) around lipid droplets in the livers of nonalcoholic steatohepatitis patients. The dimerization of HSD17B13 supports the LLPS formation and promotes its enzymatic function. HSD17B13 LLPS increases the biosynthesis of platelet activating factor (PAF), which in turn promotes fibrinogen synthesis and leukocyte adhesion. Blockade of PAFR or STAT3 pathway inhibited the fibrinogen synthesis and leukocyte adhesion. Importantly, adeno-associated viral-mediated xeno-expression of human HSD17B13 exacerbated western diet/carbon tetrachloride-induced liver inflammation in Hsd17b13-/- mice. In conclusion, our results suggest that HSD17B13 LLPS triggers liver inflammation by promoting PAF-mediated leukocyte adhesion, and targeting HSD17B13 phase transition could be a promising therapeutic approach for treating hepatic inflammation in chronic liver disease.

Acarbose a promising inhibitor of 17®-hydroxysteroid dehydrogenase type 1 as a potential candidate for anticancer therapy

Acarbose a promising inhibitor of 17®-hydroxysteroid dehydrogenase type 1 as a potential candidate for anticancer therapy

Ultrasensitive electrochemical detection of bile acids via ZIF-67-MOF-derived CoNi(OH)x/CeO2/COOH-MWCNTs composite electrodes

Fast and ultra-sensitive assays are essential for the assessment of the efficacy of traditional Chinese medicines. Here, an ultrasensitive and rapid electrochemical detection of bile acid in traditional Chinese medicine using composite electrodes is presented. Cobalt-nickel hydroxide and cerium dioxide (CoNi(OH)x/CeO2) nanomaterials were synthesized in situ by a two-step reflux method with cobalt-containing metal–organic framework (ZIF-67), as precursor. It was ultrasonically complexed with carboxylated carbon nanotubes (COOH-MWCNTs) to form CoNi(OH)x/CeO2/COOH-MWCNTs composites, for facile direct detection of bile acids in traditional Chinese medicine. Characterization methods were used to verify the successful creation of the composite CoNi(OH)x/CeO2/COOH-MWCNTs, a material which demonstrated excellent electrochemical properties. The experimental results showed that the electrode responded well when bile acids, nicotinamide adenine dinucleotide, and 3α hydroxysteroid dehydrogenase were simultaneously present in the solution, and that the peak current had a good linear relationship (R2 = 0.988) with the concentration of bile acids (0.05–10.0 µM). The sensor has an outstanding detection limit of 1.0 nM, 2–3 orders of magnitude more sensitive than the concentration. Therefore, it can be used for rapid on-site detection of bile acids in real traditional Chinese medicines.

HSD17B1 Compensates for HSD17B3 Deficiency in Fetal Mouse Testis but Not in Adults.

Hydroxysteroid (17β) dehydrogenase (HSD17B) enzymes convert 17-ketosteroids to 17beta-hydroxysteroids, an essential step in testosterone biosynthesis. Human XY individuals with inactivating HSD17B3 mutations are born with female-appearing external genitalia due to testosterone deficiency. However, at puberty their testosterone production reactivates, indicating HSD17B3-independent testosterone synthesis. We have recently shown that Hsd17b3 knockout (3-KO) male mice display a similar endocrine imbalance, with high serum androstenedione and testosterone in adulthood, but milder undermasculinization than humans. Here, we studied whether HSD17B1 is responsible for the remaining HSD17B activity in the 3-KO male mice by generating a Ser134Ala point mutation that disrupted the enzymatic activity of HSD17B1 (1-KO) followed by breeding Hsd17b1/Hsd17b3 double-KO (DKO) mice. In contrast to 3-KO, inactivation of both HSD17B3 and HSD17B1 in mice results in a dramatic drop in testosterone synthesis during the fetal period. This resulted in a female-like anogenital distance at birth, and adult DKO males displayed more severe undermasculinization than 3-KO, including more strongly reduced weight of seminal vesicles, levator ani, epididymis, and testis. However, qualitatively normal spermatogenesis was detected in adult DKO males. Furthermore, similar to 3-KO mice, high serum testosterone was still detected in adult DKO mice, accompanied by upregulation of various steroidogenic enzymes. The data show that HSD17B1 compensates for HSD17B3 deficiency in fetal mouse testis but is not the enzyme responsible for testosterone synthesis in adult mice with inactivated HSD17B3. Therefore, other enzymes are able to convert androstenedione to testosterone in the adult mouse testis and presumably also in the human testis.

Mitigating effect of pomegranate peel extract against the furan induced testicular injury by apoptosis, steroidogenic enzymes and oxidative stress

Furan is generated in a wide array of heat-treated foods through thermal degradation, leading to severe impairments in the male reproductive system. The main objective of this study was to investigate the potential of pomegranate peel extract (PGPE) in mitigating testicular dysfunctions induced by furan. Male rats were categorized into four groups: control/untreated, PGPE, furan, and PGPE + furan group. The study results revealed that furan-treated rats exhibited significantly elevated aminotransferase and phosphatase activity, and also generated increased oxidative stress, and reduced antioxidative stress protein activity. Additionally, protein content levels (ALT, AST, ALP, and ACP) and activities of steroidogenic Leydig cell hydroxysteroid dehydrogenase (3β-HSD and 17β-HSD) enzymes were significantly decreased. Significant variations in testicular parameters, apoptotic genes (Bcl-2, P53, and Caspase3), inflammatory and anti-inflammatory cytokines (IL1β, IL10), male sex hormones follicle-stimulating hormone (FSH), luteinizing hormone (LH), testosterone, and sperm quality were also observed. Furthermore, testicular histological abnormalities were confirmed by biochemical and molecular modifications. Notably, PGPE pre-treated furan-intoxicated animals exhibited significant improvements in most of the assessed parameters compared to furan-treated groups. In conclusion, PGPE presents essential preventive measures and a novel pharmacological potential therapy against furan-induced testicular injury.

A Perspective on Male Contraceptive Therapeutics of Hydro-ethanol Extract of Seeds of <i>Luffa acutangula (L.) Roxb.</i> in Human and Albino Rat: An <i>In-vitro</i> Meta-Analysis Study

Traditionally Luffa acutangula (L.) Roxb. has been utilized as an herbal contraceptive. This in-vitro study has been performed to search out the male contraceptive efficacy of hydro-ethanol (60:40) extract of Luffa acutangula (LAHEE) seeds in a dose-specific manner (1, 2, and 4 mg/ml of in-vitro media). The percentages of motile, viable, hypo-osmotic swelled (HOS), and acrosomal intact of human and rat sperms were declined significantly (p<0.05) at the above-mentioned doses of LAHEE-exposed groups against the placebo group. The inhibitory concentration 50% value (IC50) of LAHEE was 2.5 mg/ml in human and 1.2 mg/ml in rat spermatozoa immobilization. The ∆5,3β and 17β hydroxysteroid dehydrogenase (HSD) activities of rat’s testicular tissues were inhibited significantly at p<0.05 in LAHEE-treated groups than the placebo group. Activities of superoxide dismutase (SOD) and catalase were significantly inhibited (p<0.05) along with the significant increment (p<0.05) in the quantity of thiobarbituric acid reactive substances (TBARS) in rat’s testes and epididymis, sperm pellets of humans and rats in LAHEE-treated groups against the placebo group without any significant difference (p>0.05) in above said sensors in the liver and cardiac tissues of rats. The non-toxic nature of LAHEE was indicated by no significant alterations (p>0.05) in the activities of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) after LAHEE exposure in said tissues of rats. The LC-MS study of LAHEE identified the presence of oleic acid, dihydroquercetin, luteolin-7-glucoside, formononetin, luteolin 8-C-pentoside-6-C-hexoside, pterosin B, boldine and berberine. These findings interpreted that the LAHEE possesses spermiological inhibitory and hypo-testicular activities, which indicate a promising possibility to develop a potent herbal male contraceptive agent from this plant extract.

The role of cholesterol and its oxidation products in tuberculosis pathogenesis.

Mycobacterium tuberculosis causes tuberculosis (TB), one of the world's most deadly infections. Lipids play an important role in M. tuberculosis pathogenesis. M. tuberculosis grows intracellularly within lipid-laden macrophages and extracellularly within the cholesterol-rich caseum of necrotic granulomas and pulmonary cavities. Evolved from soil saprophytes that are able to metabolize cholesterol from organic matter in the environment, M. tuberculosis inherited an extensive and highly conserved machinery to metabolize cholesterol. M. tuberculosis uses this machinery to degrade host cholesterol; the products of cholesterol degradation are incorporated into central carbon metabolism and used to generate cell envelope lipids, which play important roles in virulence. The host also modifies cholesterol by enzymatically oxidizing it to a variety of derivatives, collectively called oxysterols, which modulate cholesterol homeostasis and the immune response. Recently, we found that M. tuberculosis converts host cholesterol to an oxidized metabolite, cholestenone, that accumulates in the lungs of individuals with TB. M. tuberculosis encodes cholesterol-modifying enzymes, including a hydroxysteroid dehydrogenase, a putative cholesterol oxidase, and numerous cytochrome P450 monooxygenases. Here, we review what is known about cholesterol and its oxidation products in the pathogenesis of TB. We consider the possibility that the biological function of cholesterol metabolism by M. tuberculosis extends beyond a nutritional role.

LET-767 determines lipid droplet protein targeting and lipid homeostasis.

Lipid droplets (LDs) are composed of a core of neutral lipids wrapped by a phospholipid (PL) monolayer containing several hundred proteins that vary between different cells or organisms. How LD proteins target to LDs is still largely unknown. Here, we show that RNAi knockdown or gene mutation of let-767, encoding a member of hydroxysteroid dehydrogenase (HSD), displaced the LD localization of three well-known LD proteins: DHS-3 (dehydrogenase/reductase), PLIN-1 (perilipin), and DGAT-2 (diacylglycerol O-acyltransferase 2), and also prevented LD growth in Caenorhabditis elegans. LET-767 interacts with ARF-1 (ADP-ribosylation factor 1) to prevent ARF-1 LD translocation for appropriate LD protein targeting and lipid homeostasis. Deficiency of LET-767 leads to the release of ARF-1, which further recruits and promotes translocation of ATGL-1 (adipose triglyceride lipase) to LDs for lipolysis. The displacement of LD proteins caused by LET-767 deficiency could be reversed by inhibition of either ARF-1 or ATGL-1. Our work uncovers a unique LET-767 for determining LD protein targeting and maintaining lipid homeostasis.

Abstract 3588: Androgen synthesis disruption, ferroptosis induction and prostate cancer inhibition by androgen and vitamin D combination

Abstract Background: Metastatic castration-resistant prostate cancer (CRPC) is a fatal condition that progresses to end-of-life within two years, and it is the fifth leading cause of cancer death in men globally. New avenues for preventing or delaying lethal CRPC progression are a critical unmet need. Hormonal vitamin D3 (calcitriol) inhibits prostate cancer - both castration sensitive and castration resistant - in preclinical models, as documented widely. However, vitamin D as an anti-cancer therapeutic is clinically untenable since prostate tumor suppression in patients requires hormone dosing at a supraphysiologic level which induces hypercalcemia and other toxicities. Results: Herein we provide the first evidence that a subnanomolar, non-calcemic calcitriol dose can inhibit CRPC cell proliferation, G1 to S phase transition, clonal growth, migration, and invasion in vitro, and xenograft growth in vivo, only upon concurrent dosing with 5α-DHT at a physiologic or subphysiologic level. Calcitriol as a single treatment is non-inhibitory at a subnanomolar dose. HSD3β1, a hydroxysteroid dehydrogenase, was markedly induced by calcitriol, while DHT cotreatment reduced the induction. HSD3β1 catalyzes conversion of dehydroepiandrosterone, an adrenal androgen, to androstenedione (AED) - a rate-limiting step in the androgen biosynthesis pathway. AKR1C3, an aldo-keto reductase which promotes androgen biosynthesis, was also reduced by calcitriol/DHT cotreatment. In contrast, AKR1D1, an androgen-inactivating 5β-reductase, was robustly induced by a non-calcemic calcitriol dose in the presence of DHT, not by calcitriol singly. AKR1D1 converts AED and testosterone to 5β-diols. HSD3β1 and AKR1C3 are elevated in CRPC. Disruption of androgen biosynthesis and net reduction of the intratumor androgen pool are expected outcomes of reduced HSD3β1 and AKR1C3, and elevated AKR1D1. The hormone pair led to LC3B induction, ferritin degradation, and induction of ALOX-5 and NCoA4 - the genes which regulate ferroptosis, indicating autophagy mediated ferroptosis (ferritinophagy). Apoptosis and necroptosis were absent. We conclude that vulnerabilities created by combined androgen and non-calcemic vitamin D dosing can lead to cell death by ferroptosis, and this hormone pairing may be a new avenue for arresting CRPC clinical progression. Significance: The translational potential of our results is bolstered by the precedent that supraphysiologic testosterone improved serum PSA scores and other clinical parameters for a subset of therapy-inert CRPC patients who received repeated cycles of once-a-month single testosterone dosing- a protocol known as bipolar androgen therapy (BAT)1. Our findings may offer a strategic framework for enhancing the efficacy of BAT by including oral dosing of a non-calcemic level of vitamin D3. W81XWH-21-1-0307 (DoD); IK6 BX004207 (VA). Ref 1: Nat Rev Urol 2023; PMID 36543976 Citation Format: Bobae Park, Jacqueline Vaquiz, Okunola Igbekoyi, Bandana Chatterjee. Androgen synthesis disruption, ferroptosis induction and prostate cancer inhibition by androgen and vitamin D combination [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3588.

Thiacloprid impairs reproductive functions of male Wistar rats

Global male infertility correlated to the rise of endocrine-disrupting chemicals, including insecticides, has grown into a pressing problem. Thiacloprid is one of the most commonly used neonicotinoids that accounts for more than 25% of the global pesticide industry. However, its impact on the reproductive system and male fertility has not been fully elucidated. The object of this study was to explore the adverse effects of thiacloprid on male Wistar rats’ reproductive system. Thirty healthy male rats were separated into one of three groups: control group, and two groups that were orally administered with low (22.5 mg/kg) and high dose (62.1 mg/kg) of thiacloprid for 56 days. Thiacloprid significantly (p<0.05) reduced body weight and relative testicular weight, as well as sperm quality (count, motility, viability, and morphology), in a dose-dependent manner. THIA-treated groups revealed a large effect (d > 0.8) on semen quality with Cohen’s d of (6.57, 8.82), (20.14, 23.54), and (2.81, 9.10) for count, motility, and viability respectively. Meanwhile, the serum testosterone level dropped while the levels of luteinizing and follicle-stimulating hormones increased. 17ꞵ-hydroxy steroid dehydrogenase and 3ꞵ-hydroxy steroid dehydrogenase levels were significantly decreased in a dose-dependent manner. The activity of the tested antioxidant enzymes catalase (CAT), glutathione reduced (GSH), and superoxide dismutase (SOD) exhibited a considerable decrease compared to the control group with a significant elevation in the lipid peroxidation activity as indicated by malondialdehyde (MDA) level. The testicular histology revealed degenerative changes in spermatogenic cells and interstitial tissue. Comet assay revealed DNA fragmentation in treated groups’ testicular tissue. Thiacloprid exposure interferes with reproductive function and impairs male Wistar rat fertility. Such harmful consequences may also develop in humans frequently exposed to thiacloprid.

Identification of Hydroxysteroid Dehydrogenase Type 1 As a Potential Bladder Tumor Marker.

The 17beta-hydroxysteroid dehydrogenase type 1 (HSD17B) family has been implicated in the prognosis and treatment prediction of various malignancies; however, its association with bladder cancer (BLCA) remains unclear. This study aimed to evaluate the potential of HSD17B1, as a prognostic biomarker, for the survival of patients with BLCA and to determine its effectiveness as a supplemental biomarker for BLCA. A series of bioinformatics techniques were applied to investigate the expression of HSD17B1 in different types of cancer and its potential association with the prognosis of BLCA patients using diverse databases. The UALCAN, Human Protein Atlas, cBioPortal, Metascape, GEPIA, MethSurv, and TIMER were employed to analyze expression differences, mutation status, enrichment analysis, overall survival, methylation, and immune-infiltrating cells. The real-time reverse transcription-PCR (qRT-PCR) was implemented to detect the messenger ribonucleic acid (mRNA) expression levels of HSD17B1 in vitro. Elevated mRNA and protein levels of HSD17B1, surpassing normal levels, were observed in BLCA samples. In addition, the BLCA patients with higher mRNA expression level of HSD17B1 significantly reduced the overall survival. Also, several immune infiltrating cells, including mast cell resting CIBERSORT-ABS, have been identified as tumor-associated biomarker genes, with the potential to significantly influence the immunological environment. Finally, qRT-PCR analysis revealed a significant upregulation of HSD17B1 mRNA expression level in the cancer cells compared to the human 293T cells, which was consistent with the bioinformatics data. There is a strong correlation between the elevated HSD17B1 expression and positive prognosis in patients with BLCA. Therefore, HSD17B1 can be used as a prognostic biomarker in these patients.

Another renaissance for bile acid gastrointestinal microbiology.

The field of bile acid microbiology in the gastrointestinal tract is going through a current rebirth after a peak of activity in the late 1970s and early 1980s. This renewed activity is a result of many factors, including the discovery near the turn of the century that bile acids are potent signalling molecules and technological advances in next-generation sequencing, computation, culturomics, gnotobiology, and metabolomics. We describe the current state of the field with particular emphasis on questions that have remained unanswered for many decades in both bile acid synthesis by the host and metabolism by the gut microbiota. Current knowledge of established enzymatic pathways, including bile salt hydrolase, hydroxysteroid dehydrogenases involved in the oxidation and epimerization of bile acid hydroxy groups, the Hylemon-Bjӧrkhem pathway of bile acid C7-dehydroxylation, and the formation of secondary allo-bile acids, is described. We cover aspects of bile acid conjugation and esterification as well as evidence for bile acid C3-dehydroxylation and C12-dehydroxylation that are less well understood but potentially critical for our understanding of bile acid metabolism in the human gut. The physiological consequences of bile acid metabolism for human health, important caveats and cautionary notes on experimental design and interpretation of data reflecting bile acid metabolism are also explored.

JTT-654, an 11-beta hydroxysteroid dehydrogenase type 1 inhibitor, improves hypertension and diabetic kidney injury by suppressing angiotensinogen production

11β-Hydroxysteroid dehydrogenase type 1 (11β-HSD1) plays an important role in regulating the expression of glucocorticoid actions in target tissues. Overexpression of 11β-HSD1 in mouse adipose tissue causes a metabolic syndrome-like phenotype, leading to hypertension. Although, many 11β-HSD1 inhibitors have been studied, few have shown a clear ameliorative effect against hypertension. We investigated whether JTT-654, a novel 11β-HSD1 inhibitor, ameliorated hypertension and elucidated the underlying mechanisms. JTT-654 showed inhibitory effects on angiotensinogen production in cortisone-treated 3T3-L1 adipocytes and in a rat model. JTT-654 improved hypertension not only in cortisone-treated rats and spontaneously hypertensive rats (SHR), but also in SHR/NDmcr-cp rats. In the SHR study, JTT-654 and losartan showed the same degree of antihypertensive efficacy. In addition, JTT-654 ameliorated diabetic nephropathy by suppressing renal angiotensinogen production in SHR/NDmcr-cp rats. These effects of JTT-654 were independent of its insulin-sensitizing effects, and similar effects were not observed for pioglitazone, an insulin sensitizer. Moreover, JTT-654 did not affect normotension or hypothalamus-pituitary-adrenal (HPA) axis function in normal Sprague-Dawley rats. Our results indicate that JTT-654 ameliorates hypertension and diabetic nephropathy by inhibiting 11β-HSD1 in the adipose tissue, liver, and kidney.

Evaluation of antisense oligonucleotide therapy targeting Hsd17b13 in a fibrosis mice model

Human genetic evidence suggests a protective role of loss-of-function variants in 17-beta hydroxysteroid dehydrogenase 13 (HSD17B13) for liver fibrotic diseases. Although there is limited preclinical experimental data on Hsd17b13 antisense oligonucleotide (ASO) or siRNA in a fibrosis model, several ASO and siRNA approaches are being tested clinically as potential therapies for nonalcoholic steatohepatitis (NASH). The aim of this study was to assess the therapeutic potential of Hsd17b13 ASO in a preclinical advanced NASH-like hepatic fibrosis in vivo model. In vitro testing on primary hepatocytes demonstrated that Hsd17b13 ASO exhibited strong efficacy and specificity for knockdown of the Hsd17b13 gene. In choline-deficient, L-amino acid-defined, HFD (CDAHFD)-induced steatotic and fibrotic mice, therapeutic administration of Hsd17b13 ASO resulted in a significant and dose-dependent reduction of hepatic Hsd17b13 gene expression. The CDAHFD group exhibited considerably elevated liver enzyme levels, hepatic steatosis score, hepatic fibrosis, and increased fibrotic and inflammatory gene expression, indicating an advanced NASH-like hepatic fibrosis phenotype. Although Hsd17b13 ASO therapy significantly affected hepatic steatosis, it had no effect on hepatic fibrosis. Our findings demonstrate, for the first time, that Hsd17b13 ASO effectively suppressed Hsd17b13 gene expression both in vitro and in vivo, and had a modulatory effect on hepatic steatosis in mice, but did not affect fibrosis in the CDAHFD mouse model of NASH.

QSAR of Oxazinanone Derivatives As 11-Β Hydroxysteroid Dehydrogenase Type 1 Inhibitor A Potent Anti Diabetic Agent

11-β hydroxysteroid dehydrogenase type 1 is a key enzyme accountable for the interconversion of physiologically inert cortisone to active cortisol thus presents an effective target for the development of pharmacologically active anti diabetic agents focused on manage blood glucose levels, improve insulin sensitivity. The 11β-HSD1 facilitates intracellular cortisol construction that have a disease-causing role in type 2 diabetes and the co-morbidities that it causes. Drugs in habiting the enzyme 11 β-HSD1 offers a potential therapy to lessen the type 2 diabetes. Oxazinanone ring has shown activities as antitumor, antihypertensive, antibacterial, anti-inflammatory, antioxidant and many more. Oxazinanone ring have emerged as potent inhibitors of 11β-HSD1 enzyme. QSAR of Oxazinanone derivatives is performed with a goal of elucidating the key characteristics that cause their anti-diabetic action. QSAR is the most widespread method to ligand-based drug design. It is supposed that structures of the molecules are directly proportional with biological activities, and thus, the biological activities can be altered with any structural changes. The process involves computational or mathematical models to find important correlations between a series of structures and functions. Step wise partial least square, multiple linear regressions, and feed forward neural network were used in a QSAR investigation on enzyme (IC50 nM). The developed models were cross confirmed by the ‘‘leave one out’’ method. The model reveals the significance of steric parameter Verloop B1 (Substitution 1) and Total lipole molecular descriptor.Total lipole bear a resemblance to lipophilicity which is a ratio of the capability of molecules to transfer between oily partition and aqueous partition. These descriptors will have an impact on the design and expansion of novel anti-diabetic 11-hydroxysteroid dehydrogenase type 1 inhibitors.

A Dietary Supplement Jinghuosu Ameliorates Reproductive Damage Induced by Tripterygium Glycosides.

To determine the possible protective effects of Jinghuosu, a dietary supplement (DS), on tripterygium glycosides (TG)-induced reproductive system injury in rats and its underlying mechanisms. A reproductive damage model was established in rats by feeding of TGs. Twenty-eight male Sprague Dawley rats were randomly divided into 4 groups using a random number table (n=7 in each): control (C) group, model (M) group, DS group and L-carnitine (LC) group. Rats in M, DS and LC groups received 40 mg/kg TGs orally. Starting from the 5th week, after administration of TGs for 4 h every day, rats in DS and LC groups were administered with 2.7 g/kg DS and 0.21 g/kg LC, respectively, for protective treatment over the next 4 weeks. Rats in Group C continued to receive the control treatment. Hematoxylin-eosin staining was used for histopathological analysis of rat testicular tissues. Enzyme-linked immunosorbent assay was performed to measure alkaline phosphatase (ALP), lactate dehydrogenase, alcohol dehydrogenase, total antioxidant capacity (T-AOC), superoxide dismutase, glutathione peroxidase (GSH-Px), and malondialdehyde (MDA) concentrations. Chemiluminescence assay was used to determine the serum testosterone content. Quantitative real-time PCR and Western blotting were conducted to analyze the expression of genes and proteins related to the testosterone synthesis pathway and the nuclear factor erythroid 2-related factor 2/heme oxygenase 1 antioxidant pathway. Oral administration of TGs induced significant increases in the testicular levels of zinc transporter 1 and MDA (P<0.05). On the other hand, sperm concentration, sperm motility, and serum testosterone, serum zinc, testicular zinc, Zrt-, Irt-like protein 1, ALP, luteinizing hormone (LH) receptor, steroidogenic acute regulatory protein, Cytochrome P450 family 11 subfamily A member 1, 3 β -hydroxysteroid dehydrogenase 1 T-AOC, GSH-Px, nuclear factor erythroid 2-related factor 2, heme oxygenase-1 and NAD (P)H: quinone oxidoreductase 1 levels decreased following TGs exposure (P<0.05). All of these phenotypes were evidently reversed by DS (P<0.05). DS Jinghuosu protects against TG-induced reproductive system injury in rats, probably by improving zinc homeostasis, enhancing the testosterone synthesis and attenuating oxidative stress.

Not exclusively the activity, but the sweet spot: a dehydrogenase point mutation synergistically boosts activity, substrate tolerance, thermal stability and yield.

Catalytic activity is undoubtedly a key focus in enzyme engineering. The complicated reaction conditions hinder some enzymes from industrialization even though they have relatively promising activity. This has occurred to some dehydrogenases. Hydroxysteroid dehydrogenases (HSDHs) specifically catalyze the conversion between hydroxyl and keto groups, and hold immense potential in the synthesis of steroid medicines. We underscored the importance of 7α-HSDH activity, and analyzed the overall robustness and underlying mechanisms. Employing a high-throughput screening approach, we comprehensively assessed a mutation library, and obtained a mutant with enhanced enzymatic activity and overall stability/tolerance. The superior mutant (I201M) was identified to harbor improved thermal stability, substrate susceptibility, cofactor affinity, as well as the yield. This mutant displayed a 1.88-fold increase in enzymatic activity, a 1.37-fold improvement in substrate tolerance, and a 1.45-fold increase in thermal stability when compared with the wild type (WT) enzyme. The I201M mutant showed a 2.25-fold increase in the kcat/KM ratio (indicative of a stronger binding affinity for the cofactor). This mutant did not exhibit the highest enzyme activity compared with all the tested mutants, but these improved characteristics contributed synergistically to the highest yield. When a substrate at 100 mM was present, the 24 h yield by I201M reached 89.7%, significantly higher than the 61.2% yield elicited by the WT enzyme. This is the first report revealing enhancement of the catalytic efficiency, cofactor affinity, substrate tolerance, and thermal stability of NAD(H)-dependent 7α-HSDH through a single-point mutation. The mutated enzyme reached the highest enzymatic activity of 7α-HSDH ever reported. High enzymatic activity is undoubtedly crucial for enabling the industrialization of an enzyme. Our findings demonstrated that, when compared with other mutants boasting even higher enzymatic activity, mutants with excellent overall robustness were superior for industrial applications. This principle was exemplified by highly active enzymes such as 7α-HSDH.