THERMOCHIMICA ACTA
·
2025
·
IF 3.5
Jixiu Deng,Pengshuai Zhang,Han Sun,Wang Rongrong,Binbin Wu,Yuemei Li,Beibei Feng,Haowei Sun,Shuoye Yang
This study aimed to investigate the solubility and dissolution thermodynamics of pregnenolone (Form I) in n -propanol, N-Methyl-2-pyrrolidone (NMP), ethyl acetate, 2-methoxyethanol, ethyl formate, n -butanol, 2-butoxyethanol, N, N-Dimethylformamide (DMF), 2-propoxyethanol, i -propanol, ethanol, 2-ethoxyethanol and methanol from 283.15 K to 323.15 K by a laser dynamic method. The experimental solubility of pregnenolone (Form I) was successfully fitted using five thermodynamic models (Jouyban, Apelblat, NRTL, van't Hoff, UNIQUAC), and the van't Hoff equation provided the best prediction effect with a minimum average RAD (relative average deviation) of 1.00 × 10 -2 and RMSD (root mean square deviation) of 6.58 × 10 -5 . Hansen solubility parameter (HSPs) was used to evaluate the dissolution behavior including the solubility order of pregnenolone (Form I). The apparent thermodynamics were evaluated, and results showed that entropy was the main driving force during the dissolution process of pregnenolone (Form I).
研究领域:
药学
·
物理化学
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DOI
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2018
·
IF 3.909
Di Xu,Mengyun Wu,Xue Li,Mingyu Xia,Dongchun Liu,Yinghui Dai,Qing Yu,Bin Wu,Dong Wang
Bufadienolides, one kind of steroids, are the major active component secreted by ear-side gland of Bufo species. Preliminary studies on high-throughput transcriptome sequencing about B. bufo gargarizans showed that the expression of 3 β -Hydroxysteroid dehydrogenase (3 β HSD) in ear-side gland was nearly 20 times higher than that in liver. The enzyme 3 β HSD is an essential step in the biosynthesis of steroid such as progesterone, estrogens and androgens in steroidogenic tissues. Accordingly, 3 β HSD is probably an important enzyme involved in the biosynthesis of bufadienolides. In this study, Bbg-3βHSD cDNA was cloned from the ear-side gland of B. bufo gargarizans . Genetic engineering techniques were used to construct a recombinant prokaryotic fusion expression plasmid pCOLD- Bbg3βHSD which was introduced into E. coli BL21 for prokaryotic expression. Bbg- 3 β HSD has an open reading frame (ORF) of 1134 bp and encodes 377 amino acid residues. The speculated protein molecular weight is 42.8 kDa and its theoretical isoelectric point is 8.68. Amino acid sequence homologous analysis showed that Bbg -3 β HSD was highly homologous to the 3 β HSD protein of other species. Phylogenetic tree showed the highest similarity between Bbg -3 β HSD and 3 β HSD from Rana rugosa . The optimized expression of recombinant Bbg -3 β HSD were achieved by inducing with 0.1 mmol L −1 IPTG at 15 °C for 20 h. Enzymatic activity in vitro shows that pregnenolone and dehydroepiandroesterone could be 3 β -oxidized by Bbg -3 β HSD when NAD + was used as the coenzyme. Enzymatic properties showed that the optimum reaction temperature of recombinant Bbg- 3 β HSD was 40 °C, the optimum pH was 8.5, and the optimum coenzyme concentration was 1.5 mmol L −1 .
研究领域:
生物化学
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分子生物学
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酶学
·
药理学
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产品关联
DOI
PubMed
高影响因子
ACTA PHARMACOLOGICA SINICA
·
2024
·
IF 6.9
Liang Hang-fei,Yang Xiao,Li Hui-lin,Li Xuan,Tian Jia-ning,Su Hai-guo,Huang Min,Fang Jian-hong,Bi Hui-chang
Our previous study shows that activation of pregnane X receptor (PXR) exerts hepatoprotection against lithocholic acid (LCA)-induced cholestatic liver injury. In this study we investigated whether PXR activation could inhibit hepatocyte pyroptosis, as well as the underlying mechanisms. Male mice were treated with mouse PXR agonist pregnenolone 16α-carbonitrile (PCN, 50 mg·kg −1 ·d −1 , i.p.) for 7 days, and received LCA (125 mg/kg, i.p., bid) from D4, then sacrificed 12 h after the last LCA injection. We showed that LCA injection resulted in severe cholestatic liver injury characterized by significant increases in gallbladder size, hepatocellular necrosis, and neutrophil infiltration with a mortality rate of 68%; PCN treatment significantly inhibited hepatocyte pyroptosis during LCA-induced cholestatic liver injury, as evidenced by reduced serum lactic dehydrogenase (LDH) levels, TUNEL-positive cells and hepatocyte membrane damage. Furthermore, PXR activation suppressed both the NOD-like receptor protein 3 (NLRP3) inflammasome-induced canonical pyroptosis and the apoptosis protease activating factor-1 (APAF-1) pyroptosome-induced non-canonical pyroptosis. Inhibition of the nuclear factor kappa B (NF-κB) and forkhead box O1 (FOXO1) signaling pathways was also observed following PXR activation. Notably, dual luciferase reporter assay showed that PXR activation inhibited the transcriptional effects of NF-κB on NLRP3, as well as FOXO1 on APAF-1. Our results demonstrate that PXR activation protects against cholestatic liver injury by inhibiting the canonical pyroptosis through the NF-κB-NLRP3 axis and the non-canonical pyroptosis through the FOXO1-APAF-1 axis, providing new evidence for PXR as a prospective anti-cholestatic target.
研究领域:
肝病学
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药理学
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分子生物学
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产品关联
DOI
PubMed
高影响因子
Cell Metabolism
·
2024
·
IF 27.7
Hui Lin,Chuanshun Ma,Xiao Zhuang,Shuo Liu,Dong Liu,Mingxiang Zhang,Yan Lu,Guangjian Zhou,Chao Zhang,Tengwei Wang,Zihao Zhang,Lin Lv,Daolai Zhang,Xiong-Zhong Ruan,Yunfei Xu,Renjie Chai,Xiao Yu,Jin-Peng Sun,Bo Chu
G protein-coupled receptors (GPCRs) mediate most cellular responses to hormones, neurotransmitters, and environmental stimulants. However, whether GPCRs participate in tissue homeostasis through ferroptosis remains unclear. Here we identify that GPR56/ADGRG1 renders cells resistant to ferroptosis and deficiency of GPR56 exacerbates ferroptosis-mediated liver injury induced by doxorubicin (DOX) or ischemia-reperfusion (IR). Mechanistically, GPR56 decreases the abundance of phospholipids containing free polyunsaturated fatty acids (PUFAs) by promoting endocytosis-lysosomal degradation of CD36. By screening a panel of steroid hormones, we identified that 17α-hydroxypregnenolone (17-OH PREG) acts as an agonist of GPR56 to antagonize ferroptosis and efficiently attenuates liver injury before or after insult. Moreover, disease-associated GPR56 mutants were unresponsive to 17-OH PREG activation and insufficient to defend against ferroptosis. Together, our findings uncover that 17-OH PREG-GPR56 axis-mediated signal transduction works as a new anti-ferroptotic pathway to maintain liver homeostasis, providing novel insights into the potential therapy for liver injury.
研究领域:
药理学
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细胞生物学
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分子生物学
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肝病学
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DOI
PubMed
高影响因子
TOXICOLOGY LETTERS
·
2023
·
IF 3.5
Yingna Zhai,Shaowei Wang,Bingru Zhang,Yunbing Tang,Hong Wang,Jingjing Li,Zhiyan Hu,Yiyan Wang,Huitao Li,Ren-shan Ge
3β-Hydroxysteroid dehydrogenase/steroid Δ 5,4 -isomerase 1 (3β-HSD1) plays a critical role in the biosynthesis of progesterone from pregnenolone in the human placenta to maintain normal pregnancy. Whether they inhibit placental 3β-HSD1 and mode of inhibition remains unclear. In this study, we screened 21 pesticides and fungicides in five classes to inhibit human 3β-HSD1 and compared them to rat homolog 3β-HSD4. 3β-HSD activity was measured by catalyzing pregnenolone to progesterone in the presence of NAD + . Of the 21 chemicals, azoles (difenoconazole), thiocarbamates (thiram and ferbam) and organochlorine (hexachlorophene) significantly inhibited human 3β-HSD1 with half maximal inhibitory concentration (IC 50 ) values of 2.77, 0.24, 0.68, and 17.96 μM, respectively. We also found that difenoconazole, ferbam and hexachlorophene are mixed/competitive inhibitors of 3β-HSD1 while thiram is a mixed/noncompetitive inhibitor. Docking analysis showed that difenoconazole and hexachlorophene bound steroid-binding site. Difenoconazole and hexachlorophene except thiram and ferbam also significantly inhibited rat 3β-HSD4 activity with IC 50 of 1.12 and 2.28 µM, respectively. Thiram and ferbam significantly inhibited human 3β-HSD1 possibly by interfering with cysteine residues, while they had no effects on rat 3β-HSD4. In conclusion, some pesticides potently inhibit placental 3β-HSD, leading to the reduction of progesterone formation.
研究领域:
药理学
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毒理学
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生殖生物学
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酶学
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产品关联
DOI
PubMed
高影响因子
DRUG METABOLISM AND DISPOSITION
·
2024
·Vol. 52
·期 7
·597-605
·
IF 4.4
Ting Wu,Lu Li,Wenhong Zhou,Guofang Bi,Xiaowen Jiang,Manlan Guo,Xiao Yang,Jianhong Fang,Jianxin Pang,Shicheng Fan,Huichang Bi
Pregnane X receptor (PXR) is essential in the regulation of liver homeostasis, and the gut microbiota is closely linked to liver physiologic and pathologic status. We previously found that activation of PXR significantly promotes liver enlargement through interaction with yes-associated protein (YAP). However, whether gut microbiota contributes to PXR-induced hepatomegaly and the involved mechanisms remain unclear. In this study, C57BL/6 mice were administered the mouse-specific agonist pregnenolone 16α-carbonitrile (PCN) for 5 days. Depletion of gut microbiota was achieved using broad-spectrum antibiotics (ABX) and fecal microbiota transplantation (FMT) was performed to restore the gut microbia. The composition of gut microbiota was analyzed by 16S rRNA sequencing, while the expression of PXR, YAP, and their downstream target genes and proteins were assessed. The results indicated that PCN treatment altered the composition and abundance of specific bacterial taxa. Furthermore, depletion of gut microbiota using ABX significantly attenuated PCN-induced hepatomegaly. FMT experiments further demonstrated that the fecal microbiota from PCN-treated mice could induce liver enlargement. Mechanistic studies revealed that ABX treatment impeded the PXR and YAP activation induced by PCN, as evidenced by decreased expression of PXR, YAP, and their downstream targets. Moreover, alterations in PXR and YAP activation were likely contributing to hepatomegaly in recipient mice following FMT from PCN-treated mice. Collectively, the current study demonstrated that gut microbiota is involved in PCN-induced hepatomegaly via regulating PXR and YAP activation, providing potential novel insights into the involvement of gut microbiota in PXR-mediated hepatomegaly.
研究领域:
药理学
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微生物学
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肝脏病学
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分子生物学
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DOI
PubMed
高影响因子
STEROIDS
·
2022
·
IF 2.76
Yaqin Jia,Zhe Wang,Yuyi Feng,Meixian Wang,Lili Jiang,Zhijun Yu,Xiaoguang Shao,Guiyuan He,Yong Liu
Quantification of endogenous steroids and their precursors is essential for diagnosis of a wide range of causes for female infertility. However, immunoassays often overestimate concentrations due to assay interference by other endogenous steroids, especially at low concentrations. In addition, it still lacks of diagnostic reference intervals for five sex steroid hormones, including estradiol (E 2 ), 11-deoxycorticosterone (DOC), 17-hydroxyprogesterone (17-OHP 4 ), pregnenolone (P 5 ) and progesterone (P 4 ), which are crucial for distinguishing between normal individuals and female infertility. Therefore, we developed and validated a reliable and rapid ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for simultaneous determination and quantification of five sex hormones, giving the reference intervals to accurately evaluate and diagnose female infertility. Our results showed that the developed UPLC-MS/MS assay was fast, high throughput, reproducible, specific, accurate, highly sensitive, and fully validated for simultaneous determination of P 5 , P 4 , 17-OHP 4 , DOC and E 2 in human follicular fluid. The simple sample preparation procedure in the current study gave reproducible and consistent recoveries. The validation results show that the UPLC-MS/MS assay has acceptable accuracy and precision at low concentrations, which permits their use in clinical study. In addition, our data gave the concentration range of five steroid hormones quantification in patients with female infertility and normal individuals. Our data can be used to accurately evaluate and diagnose female infertility.
研究领域:
生殖医学
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内分泌学
·
临床化学
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产品关联
DOI
PubMed
高影响因子
ACS Synthetic Biology
·
2023
·
IF 4.7
Ruosi Zhang,Mingdong Yao,Haidi Ma,Wenhai Xiao,Ying Wang,Yingjin Yuan
Substrate competition within a metabolic network constitutes a common challenge in microbial biosynthesis system engineering, especially if indispensable enzymes can produce multiple intermediates from a single substrate. Androstenedione (4AD) is a central intermediate in the production of a series of steroidal pharmaceuticals; however, its yield via the coexpression of 3β-hydroxysteroid dehydrogenase (3β-HSD) and 17α-hydroxylase/17,20-lyase (CYP17A1) in a microbial chassis affords a nonlinear pathway in which these enzymes compete for substrates and produce structurally similar unwanted intermediates, thereby reducing 4AD yields. To avoid substrate competition, we split the competing 3β-HSD and CYP17A1 pathway components into two separate Yarrowia lipolytica strains to linearize the pathway. This spatial segregation increased substrate availability for 3β-HSD in the upstream strain, consequently decreasing the accumulation of the unwanted intermediate 17-hydroxypregnenolone (17OHP5) from 94.8 ± 4.4% in single-chassis monocultures to 24.8 ± 12.6% in cocultures of strains expressing 3β-HSD and CYP17A1 separately. Orthologue screening to increase CYP17A1 catalytic efficiency and the preferential production of desired intermediates increased the biotransformation capacity in the downstream pathway, further decreasing 17OHP5 accumulation to 3.9%. Furthermore, nitrogen limitation induced early 4AD accumulation (final titer, 7.71 mg/L). This study provides a framework for reducing intrapathway competition between essential enzymes during natural product biosynthesis as well as a proof-of-concept platform for linear steroid production.
研究领域:
代谢工程
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生物技术
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药学
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微生物生物技术
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产品关联
DOI
PubMed
高影响因子
CHEMISTRY & BIODIVERSITY
·
2022
·Vol. 19
·期 10
·e202200648
·
IF 2.745
Xiao-San Li,Mao-Xun Yang,Yu-Hao Luo,He-Hui Zhan,Mei-Fang Chen,Yong-Mei Huang,Li Liu,Xue-Mei Yang
A series of novel D-ring fused or substituted steroidal N-heterocycles were synthesized, and their chemical structures were characterized by spectroscopic analysis. The anticancer activity of these compounds against four human cancer cell lines (MCF-7, H1299, HeLa and HepG2) were evaluated and the structure-activity relationship (SAR) was also investigated. Compound 3c displayed significant inhibitory activity on the four cancer cells with IC 50 values ranging from 3.88 to 10.05 μM. Overall, these studies indicated that construction of N-heterocyclic system with D-ring substituted containing a double bond at C-16 and C-17 or D-ring fused with [17,16-d]azolo[1,5-a]pyrimidine could be a promising strategy to improve antitumor activity for steroids deserved further investigation. Graphical
研究领域:
有机化学
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药物化学
·
肿瘤学
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产品关联
DOI
PubMed
高影响因子
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