Frontiers in Pharmacology
·
2024
·
IF 4.4
Chu Simeng,Yang Wei,Lu Yujie,Li Junjie,Peng Jiamin,Liu Wenjuan,Jiang Min,Bai Gang
Introduction: Tetrandrine (Tet) is the main pharmacological component of Stephania tetrandra S. Moore, which is a well-documented traditional Chinese medicine known for its diuretic and antihypertensive properties. Unraveling the specific targets and mechanisms of Tet involved in inducing diuresis and mitigating hypertension can provide valuable insights into its therapeutic effects. This study aimed to explore the diuretic and antihypertensive targets and mechanisms of Tet using chemical biology coupled with activity analyses in vivo and in vitro.Methods: The diuretic effects of Tet were evaluated using a water-loaded mouse model. The direct target proteins for the diuretic and antihypertensive effects of Tet were determined using chemical biology. Furthermore, the molecular mechanism of Tet binding to target proteins was analyzed using a multidisciplinary approach based on the structure and function of the proteins. Finally, the effects of the Tet-targeted protein on downstream signaling pathways and blood pressure were evaluated in hypertensive model rats.Results: Tet exhibited significant antihypertensive and potassium-preserving diuretic effects. The mechanism underlying these effects involves the modulation of the enzyme activity by covalent binding of Tet to Cys423 of CYP11A1. This interaction alters the stability of heme within CYP11A1, subsequently impeding electron transfer and inhibiting aldosterone biosynthesis.Discussion: This study not only revealed the mechanism of the diuretic and antihypertensive effects of Tet but also discovered a novel covalent inhibitor of CYP11A1. These findings contribute significantly to our understanding of the therapeutic potential of Tet and provide a foundation for future research in the development of targeted treatments for hypertension.
研究领域:
药理学
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化学生物学
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心血管研究
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分子生物学
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DOI
PubMed
高影响因子
METABOLIC ENGINEERING
·
2026
·
IF 7.3
Xuqian Fan,Rongxin Zhang,Gen Li,Kaiyu Ma,Qilong Wen,Feiyu Fan,Xueli Zhang
Vertebrate CYP11A1 gains activity in yeast through coupling with a Class II-type electron transfer system (ETS). • AI-guided engineering identifies the SsCYP11A1 F75S mutation that enhances SsCYP11A1–AoCPR interaction and activity. • Disruption of the vacuolar iron transporter CCC1 enhances the function of Class I-type ETS in the yeast cytosol • CYP11A1 utilizes dual ETSs to drive gram-scale de novo progesterone biosynthesis in yeast. CYP11A1-mediated catalysis is the rate-limiting step in the de novo biosynthesis of progesterone in yeast, and its relatively low catalytic efficiency largely results from insufficient electron transfer efficiency. In this study, we addressed this bottleneck by systematically reconstructing the electron transfer system (ETS) of CYP11A1. We found that, beyond its native Class I-type ETS (Adx/AdR), CYP11A1 can also function with diverse Class II-type ETSs (CPRs) across species. Screening identified the combination of CYP11A1 from Sus scrofa (SsCYP11A1) and CPR from Absidia orchidis (AoCPR) as the most effective pair. Furthermore, fusion of the two proteins via a flexible GGGS linker significantly enhanced catalytic efficiency. Subsequently, single-site mutational scanning of SsCYP11A1 using the protein language model ESM-2 identified the F75S variant, which increased progesterone titer by 2.43-fold compared to the wild type. Further analysis integrating residue flexibility prediction and site-directed mutagenesis revealed that the F75S mutation improves overall catalytic efficiency by strengthening interfacial interactions between SsCYP11A1 and AoCPR and stabilizing a catalytically favorable conformation. In addition, deletion of the vacuolar iron transporter CCC1 increased the availability of [2Fe-2S] clusters in the yeast cytosol, thereby enhancing Adx functionality and enabling the coordinated operation of Class I-type and Class II-type ETSs. Ultimately, in a 5 L bioreactor fermentation system, the engineered yeast chassis achieved a progesterone titer of 1594.15 ± 71.36 mg/L, representing the highest reported level of de novo progesterone production in yeast to date.
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DOI
高影响因子
Veterinary Sciences
·
2025
·Vol. 12
·期 5
·428
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IF 2.3
Yanbing Lou,Tingting Yang,Chenglong Xia,Shijun Yang,Huidan Deng,Yanqiu Zhu,Jing Fang,Zhicai Zuo,Hongrui Guo
Simple SummaryYak, as a unique species of plateau cattle, is highly susceptible to Cu deficiency, which is further exacerbated by poor soil conditions and low nutrient content in yak feed, which is associated with the decline of yak performance. Hormones synthesized by granulosa cells of yak follicles play an indispensable role in reproductive function. In this study, we investigated the effects of different concentrations of copper on the hormone secretion of cultured granulosa cells of yak follicles and the related molecular mechanisms. The above results showed that copper could positively promote the hormone secretion of female yaks, which provided a strategy direction with potential value for improving the fertility of yaks under the condition of copper deficiency.Yak (Bos grunniens) is the main economic animal growing in the Qinghai–Tibet Plateau. Because of its poor growing environment, copper deficiency is common. Studies have shown that appropriate copper supplementation can improve the reproductive performance of heifers. In this study, the primary granulosa cells of yak were isolated, and different copper levels were established by adding copper chelator and copper sulfate. Biochemical, Western blot and transcriptome methods were used to reveal the mechanism of copper on the hormone secretion function of granulosa cells. The results showed that copper promoted hormone secretion by regulating the expression of steroid synthase such as StAR, CYP19A1 and CYP11A1. Transcriptome analysis showed that copper ion levels significantly affected the function of cells, and changes in copper ion level affected genes related to protein phosphorylation, lipid metabolism, lipid biosynthesis, steroid hormone synthesis and the MAPK pathway. In conclusion, copper can promote steroid hormone synthesis in the granulosa cells of yak follicles, and the MAPK signaling pathway may be involved.
研究领域:
动物科学
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生殖生物学
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分子生物学
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营养学
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DOI
PubMed
高影响因子
Bioresources and Bioprocessing
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2022
·Vol. 9
·期 1
·1-14
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IF 4.983
Liu Kun,Wang Feng-Qing,Liu Ke,Zhao Yunqiu,Gao Bei,Tao Xinyi,Wei Dongzhi
Progesterone is one of the classical hormone drugs used in medicine for maintaining pregnancy. However, its manufacturing process, coupled with organic reagents and poisonous catalysts, causes irreversible environmental pollution. Recent advances in synthetic biology have demonstrated that the microbial biosynthesis of natural products, especially difficult-to-synthesize compounds, from building blocks is a promising strategy. Herein, overcoming the heterologous cytochrome P450 enzyme interdependency in Mycolicibacterium neoaurum successfully constructed the CYP11A1 running module to realize metabolic conversion from waste phytosterols to progesterone. Subsequently, progesterone yield was improved through strategies involving electron transfer and NADPH regeneration. Mutant CYP11A1 (mCYP11A1) and adrenodoxin reductase (ADR) were connected by a flexible linker (L) to form the chimera mCYP11A1-L-ADR to enhance electron transfer. The chimera mCYP11A1-L-ADR, adrenodoxin (ADX), and ADR-related homolog ARH1 were expressed in M. neoaurum, showed positive activity and produced 45 mg/L progesterone. This electron transfer strategy increased progesterone production by 3.95-fold compared with M. neoaurum expressing mCYP11A1, ADR, and ADX. Significantly, a novel inorganic–biological hybrid system was assembled by combining engineered M. neoaurum and InP nanoparticles to regenerate NADPH, which was increased 84-fold from the initial progesterone titer to 235 ± 50 mg/L. In summary, this work highlights the green and sustainable potential of obtaining synthetic progesterone from sterols in M. neoaurum.
研究领域:
合成生物学
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生物技术
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环境科学
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药学
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微生物学
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产品关联
DOI
PubMed
高影响因子
ENVIRONMENTAL TOXICOLOGY
·
2023
·
IF 4.5
Peipei Pan,Zina Wen,Feifei Ma,Zhen Lei,Chengshuang Pan,Qianjin Fei,Erpo Tian,Yiyan Wang,Qiqi Zhu,Huitao Li,Xiaoheng Li,Ying Zhong,Ren-Shan Ge,Ren-ai Xu
Bisphenol S (BPS) is a novel bisphenol A (BPA) analogue, a ubiquitous environmental pollutant that disrupts male reproductive system. Whether BPS affects Leydig cell maturation in male puberty remains unclear. Male Sprague–Dawley rats (age of 35 days) were daily gavaged to 0, 1, 10, 100, and 200 mg/kg/day from postnatal days 35–56. BPS at 1–10 mg/kg/day and higher doses markedly reduced serum testosterone and progesterone levels but it at 200 mg/kg/day significantly increased estradiol level. BPS at 100 and 200 mg/kg/day significantly elevated serum luteinizing hormone (LH) levels. BPS at 1–10 mg/kg/day and higher doses significantly reduced inhibin A and inhibin B levels. BPS at 100 and 200 mg/kg/day markedly increased CYP11A1 + Leydig cell number, but did not affect HSD11B1 + (a mature Leydig cell marker) cell number. BPS at 10 mg/kg/day and higher doses significantly downregulated the expression of Cyp11a1 and at 100 and 200 mg/kg/d significantly lowered Cyp17a1 , Hsd11b1 , and Nr5a1 in the testes. BPS at 100 and/or 200 mg/kg/day significantly elevated Lhb in the pituitary. BPS at 100 and 200 mg/kg/day significantly increased the phosphorylation of AKT1, AKT2, and CREB without affecting total AKT1, AKT2, and CREB levels. BPS at 1–100 μM significantly suppressed testosterone production and induced proliferation of primary immature Leydig cells after 24 h of treatment and these actions were reversed by estrogen receptor α antagonist, ICI 182780, and partially reversed by vitamin E. BPS at 0.1–10 μM significantly increased oxidative stress of Leydig cells in vitro. BPS also directly inhibited 17β-hydroxysteroid dehydrogenase 3 activity at 10–100 μM. In conclusion, BPS causes hypergonadotropic androgen deficiency in male rats during pubertal exposure via activating ESR1 and inducing ROS in immature Leydig cells and directly inhibiting 17β-hydroxysteroid dehydrogenase 3 activity.
研究领域:
生殖生物学
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内分泌学
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毒理学
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环境健康
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产品关联
DOI
PubMed
高影响因子
ECOTOXICOLOGY AND ENVIRONMENTAL SAFETY
·
2024
·
IF 6.2
Ling-Zi Xia,Li-Lan Liu,Jun-Zhe Yue,Zhen-Yu Lu,Jie Zheng,Ming-Zhe Jiang,Min Lin,Jiaming Liu,Hai-Tao Gao
Phthalates (PEs) are widely used plasticizers in polymer products, and humans are increasingly exposed to them. This study was designed to investigate the alleviative effect of phytochemicals quercetin (Que) against male reproductive toxicity caused by the mixture of three commonly used PEs (MPEs), and further to explore the underlying mechanism. Forty-eight male SD rats were randomly and evenly divided into control group, Que group, MPEs group and MPEs+Que group ( n = 12); The oral exposure doses of MPEs and Que were 450 mg/kg/d and 50 mg/kg/d, respectively. After 91 days of continuous intervention, compared with control group, the testes weight, epididymis weight, serum sex hormones, and anogenital distance were significantly decreased in MPEs group ( P < 0.05); Testicular histopathological observation showed that all seminiferous tubules were atrophy, leydig cells were hyperplasia, spermatogenic cells growth were arrested in MPEs group. Ultrastructural observation of testicular germ cells showed that the edges of the nuclear membranes were indistinct, and the mitochondria were severely damaged with the cristae disrupted, decreased or even disappeared in MPEs group. Immunohistochemistry and Western blot analysis showed that testicular CYP11A1, CYP17A1 and 17β-HSD were up-regulated, while StAR, PIWIL1 and PIWIL2 were down-regulated in MPEs group ( P < 0.05); However, the alterations of these parameters were restored in MPEs+Que group. The results indicated MPEs disturbed steroid hormone metabolism, and caused male reproductive injuries; whereas, Que could inhibit MPEs’ male reproductive toxicity, which might relate to the restored regulation of steroid hormone metabolism.
研究领域:
毒理学
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药理学
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生殖生物学
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内分泌学
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产品关联
DOI
PubMed
高影响因子
ENVIRONMENTAL TOXICOLOGY
·
2023
·
IF 4.5
Shuang Wang,Xueyan Gu,Jianhua Ma,Zhangyu Gu,Rui Zhang,Ruifen Li,Jun Bai,Peng Li,Linyu Wei,Yixing Ye,Yan Wang,Li Zhang,Li Su,Changhao Liang
The present study was designed to investigate the role of miR-708-5p/p38 mitogen-activated protein kinase (MAPK) pathway during the mechanism of selenium nanoparticles (Nano-Se) against nickel (Ni)-induced testosterone synthesis disorder in rat Leydig cells. We conducted all procedures based on in vitro culture of rat primary Leydig cells. After treating Leydig cells with Nano-Se and NiSO 4 alone or in combination for 24 h, we determined the cell viability, reactive oxygen species (ROS) levels, testosterone production, and the protein expression of key enzymes involved in testosterone biosynthesis: steroidogenic acute regulatory (StAR) and cytochrome P450 cholesterol side chain cleavage enzyme (CYP11A1). The results indicated that Nano-Se antagonized cytotoxicity and eliminated ROS generation induced by NiSO 4 , suppressed p38 MAPK protein phosphorylation and reduced miR-708-5p expression. Importantly, we found that Nano-Se upregulated the expression of testosterone synthase and increased testosterone production in Leydig cells. Furthermore, we investigated the effects of p38 MAPK and miR-708-5p using their specific inhibitor during Nano-Se against Ni-induced testosterone synthesis disorder. The results showed that Ni-inhibited testosterone secretion was alleviated by Nano-Se co-treatment with p38 MAPK specific inhibitor SB203580 and miR-708-5p inhibitor, respectively. In conclusion, these findings suggested Nano-Se could inhibit miR-708-5p/p38 MAPK pathway, and up-regulate the key enzymes protein expression for testosterone synthesis, thereby antagonizing Ni-induced disorder of testosterone synthesis in Leydig cells.
研究领域:
分子生物学
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内分泌学
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毒理学
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DOI
PubMed
高影响因子
Journal of Animal Science and Biotechnology
·
2022
·Vol. 13
·期 1
·1-13
·
IF 6.175
Zhou Xiaoge,Mo Zhaoyi,Li Yankun,Huang Liang,Yu Sihai,Ge Lan,Hu Yamei,Shi Shengjie,Zhang Lutong,Wang Liguang,Gao Lei,Yang Gongshe,Chu Guiyan
Background Oleic acid is an abundant free fatty acid present in livestock that are in a negative energy-balance state, and it may have detrimental effects on female reproduction and fertility. Oleic acid induces lipid accumulation in bovine granulosa cells, which leads to a foam cell-like morphology and reduced steroidogenesis. However, why oleic acid increases lipid accumulation but decreases steroidogenesis remains unclear. This study focused on oleic acid’s effects on lipid type and steroidogenesis. Results Oleic acid increased the lipid accumulation in a concentration-dependent manner and mainly increased the triglyceride level and decreased the cholesterol ester level. Oleic acid also led to a decline in estradiol and progesterone production in porcine granulosa cells in vitro. In addition, oleic acid up-regulated the expression of CD36 and diacylglycerol acyltransferase 2, but down-regulated the expression of 3-hydroxy-3-methylglutaryl-coenzyme A reductase, scavenger receptor class B member 1 and acetyl-Coenzyme A acetyltransferase 2, as well as steroidogenesis-related genes, including cytochrome P450 family 11 subfamily A member 1, cytochrome P450 family 19 subfamily A member 1 and 3 as well as steroidogenic acute regulatory protein at the mRNA and protein levels. An oleic acid-rich diet also enhanced the triglyceride levels and reduced the cholesterol levels in ovarian tissues of female mice, which resulted in lower estradiol levels than in control-fed mice. Compared with the control, decreases in estrus days and the numbers of antral follicles and corpora lutea, as well as an increase in the numbers of the atretic follicles, were found in the oleic acid-fed female mice. Conclusions Oleic acid changed the lipid type stored in lipid droplets of ovarian granulosa cells, and led to a decrease in steroidogenesis. These results improve our understanding of fertility decline in livestock that are in a negative energy-balance state.
研究领域:
生殖生物学
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内分泌学
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动物科学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）