MOLECULES
·
2016
·Vol. 21
·期 3
·349
·
IF 2.465
Ying-Hui Dai,Bo Shen,Ming-Yu Xia,An-Dong Wang,Yu-Lin Chen,Dong-Chun Liu,Dong Wang
A new indole alkaloid named bufobutarginine (1), along with three known bufotenines, namely, serotonin (2), bufotenidine (3), and bufotenine (4), were isolated from the water extract of toad venom. Their structures were elucidated by spectral methods. This is the first time that arginine has been found to be involved in the biosynthesis of bufotenines in parotid of toad. The cytotoxic activities of these compounds have been assayed against A375 and A549 cell lines by the MTT method; however, they showed no cytotoxic activities.Keywords:Bufo bufo gargarizans;toad venom;bufobutarginine;cytotoxic activity
研究领域:
药理学
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生物化学
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DOI
PubMed
高影响因子
AQUATIC ECOLOGY
·
2018
·Vol. 53
·期 1
·9-19
·
IF 1.978
Zhao Tian,Wang Xiaoyi,Wang Xungang,Wang Sishuo,Chen Youhua,Jiang Jianping
As one of the important contributors of biodiversity, amphibian populations are declining worldwide. Numerous factors are involved in these declines, one of them being the use of fertilizers in agriculture. This is especially true for tadpoles which can live in the fertilizer-polluted farmland water bodies until metamorphosis. The present study aimed to assess the effects of urea (CH 4 N 2 O), as one of the most economical and effective fertilizers, on the anti-predator behavior and intraspecific functional trait variability of Asiatic toad ( Bufo gargarizans ) tadpoles. Based on published literatures and the field observation of urea concentrations in China, glass beakers with a gradient of urea concentrations (0, 200, 400, 600, and 1200 mg/L) were prepared, with 10 tadpoles placed in each glass beaker. Each treatment was replicated three times. Mosquito fish ( Gambusia affini s) cues were used as the predator disturbance, and three main functional traits (body mass, trunk bending shape, and eye position) were selected. Our results revealed that tadpoles activity levels decreased when exposed to urea as well as to mosquito fish cues. However, urea exposure did not alter the anti-predator behaviors of tadpoles. Additionally, we found that increasing urea concentrations might modify some functional traits of tadpoles. Importantly, urea disturbance decreased tadpoles intraspecific functional trait variability. (Functional similarity increased between developmental stages.) Given that functional similarity between developmental stages could potentially increase intraspecific competition, urea could indirectly reduce tadpoles survival by decreasing intraspecific traits variability.
研究领域:
生态学
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环境科学
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DOI
高影响因子
JOURNAL OF CHROMATOGRAPHY A
·
2022
·
IF 4.601
Qianying Sheng,Ling Wang,Leyuan Zhang,Xue Wang,Shengxu Qian,Minbo Lan,Guangyan Qing,Xinmiao Liang
Natural products are rather complex samples containing a large number of compounds ranging from polar to nonpolar, small molecules to macromolecules , as well as numerous homologs/analogs with quite similar structures, which create great opportunities and treasures for discovery of bioactive drugs. For the purpose of better understanding the complex natural products and controlling their qualities, powerful analytical techniques for adequately separating chemical constituents and tracking potentially bioactive components are quite essential. Here, a design concept of bidirectional β-cyclodextrin (β-CD)–modified chromatographic stationary phase toward separation of bufadienolides extracted from toad is proposed. Bufadienolides could be divided into two classes: toad venom ligand (AAUBs) and toad toxin (AACBs) with remarkable differences in structures and polarity. The hydrophobic cavity of β-CD can encapsulate the steroid part of AACBs while the hydroxyls exposed on the β-CD surface have strong hydrophilic interactions with the arginine part of AACBs. Isothermal titration calorimetry and hydrogen nuclear magnetic resonance titration experiment further validate the rationality of this design. Furthermore, the β-CD-based stationary phase can be used as a hydrophilic material to construct a HILIC × RPLC 2D separation mode for the separation of AACBs, also works as a reverse phase material to construct a RPLC × RPLC 2D separation mode for the separation of AAUBs with good orthogonality. This study will open an avenue and provide a novel insight for high-efficiency two-dimensional separation of natural products.
研究领域:
分析化学
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药物化学
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DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2024
·
IF 7.7
Yancheng Xie,Xue Feng,Jiaqi Tao,Qianhui Gao,Yue Li,Xinyi Liu,Mingyu Xia,Dong Wang
5-Hydroxytryptamine (5-HT) and its derivative bufotenine, which possess important physiological functions, are the primary active components in the secretions of toad parotid and skin gland. However, the biosynthetic pathway of these substances remains unclear in toads. To characterize toad's Aromatic-L-amino-acid decarboxylase (AADC), the key enzyme in the predicted 5-HT derivatives biosynthetic pathway, the full-length cDNA of AADC from Bufo bufo gargarizans ( Bbg AADC) was cloned from the parotoid gland of B. bufo gargarizans . The recombinant Bbg AADC exhibited optimal expression in E. coli BL21 (DE3) containing pCold- Bbg AADC after induction for 16 h at 15 °C with 0.3 mM IPTG, resulting in substantial yields of soluble proteins . The enzymological properties of Bbg AADC were assessed, and it was determined that the optimal reaction temperature was 37 °C, the optimal pH was 8.6, and the optimum molar ratio of pyridoxal-5′-phosphate (PLP) to Bbg AADC was found to be 3.6:1. Additionally, high substrate specificity was observed, as Bbg AADC could catalyze the production of 5-HT from 5-hydroxytryptophan (5-HTP) but not dopamine or tryptamine from levodopa or tryptophan , respectively. The Km of the recombinant protein Bbg AADC was 0.2918 mM and the maximum reaction rate (Vmax) was 1.182 μM·min −1 when 5-HTP was used as substrate. The Kcat was 0.0545 min −1 , and Kcat/Km was 0.1868 mM −1 ·min −1 . To elucidate the mechanism of Bbg AADC, molecular docking was performed with PLP and 5-HTP, or the external aldimine formed by 5-HTP and PLP. The results indicated that the active sites for Bbg AADC to bind with PLP were K303, H192, N300, A148, F309, T246, A273, and T147. W71, Y79, F80, P81, T82, H192, T246, N300, H302, F309, and R477 served as catalytically active sites for the binding of Bbg AADC to 5-HTP. Furthermore, R447, W71, S149, N300, A148, and T147 of Bbg AADC were involved in the decarboxylation reaction of the aldimine formed by PLP and 5-HTP.
研究领域:
生物化学
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分子生物学
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酶学
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DOI
PubMed
高影响因子
MOLECULES
·
2020
·Vol. 25
·期 18
·4269
·
IF 3.267
Ji-Heng Wu,Yue-Ting Cao,Hong-Ye Pan,Long-Hu Wang
(1) Background: Toad venom (Bufonis Venenum, known as ‘Chansu’ in Chinese), the secretion of the ear-side gland and skin gland of Bufo gargarizans cantor or Duttaphrynus melanostictus Schneider, has been utilized to treat several diseases in China for thousands of years. However, due to the chemical variability of the components, systematic chemical composition and the key pharmacophores in toad venom have not yet fully understood. Besides, it contains a variety of effective compounds with different physiological activity and chemotypes, mainly including alkaloids, bufogenins, bufotoxins, and so on. The recent pharmacological researches have demonstrated that several bufogenins have remarkable pharmacological effects, such as anti-inflammatory, analgesic effects, and anti-tumor effects. Aim of the study: To identify the bioactive compounds and pharmacophores originating from toad venom based on analyzing spectrum-effect relationship by chemometrics and to explore the anti-cancer mechanism primarily. (2) Materials and methods: Fingerprint of the 21 batches of samples was established using HPLC (High Performance Liquid Chromatography). The anti-tumor activity of extracts were determined by in-vitro assays. Chemometric analysis was used to establish the spectrum-effect model and screen for active ingredients. Pharmacodynamic tests for the screened active compound monomers were conducted with in-vitro assays. Further anti-tumor mechanisms were investigated using western blot and flow cytometry. (3) Results: The established spectrum-effect model has satisfactory fitting effect and predicting accuracy. The inhibitory effect of major screened compounds on lung carcinoma cells A549 were validated in vitro, demonstrating that arenobufagin, telocinobufogenin, and cinobufotalin had significant anti-tumor effects. Through further investigation of the mechanism by western blotting and flow cytometry, we elucidated that arenobufagin induces apoptosis in A549 cells with the enhanced expression of cleaved PARP (poly (ADP-ribose) polymerase). These results may provide valuable information for further structural modification of bufadienolides to treat lung cancer and a method for discovery of anti-tumor active compounds. Conclusions: Our research offers a more scientific method for screening the principal ingredients dominating the pharmacodynamic function. These screened compounds (arenobufagin, etc.) were proven to induce apoptosis by overactivation of the PARP-pathway, which may be utilized to make BRCA (breast cancer susceptibility gene) mutant cancer cells more vulnerable to DNA damaging agents and kill them.Keywords:toad venom;spectrum-effect relationship;chemometrics;apoptosis
研究领域:
药理学
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肿瘤学
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化学计量学
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分子生物学
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DOI
PubMed
高影响因子
PHARMACEUTICAL DEVELOPMENT AND TECHNOLOGY
·
2023
·
IF 3.4
Quandong Li,Xinru Chen,Wen Lin,Xiaofeng Guo,Yan Ma
Toad skin has many pharmacological activities and bufadienolides are regarded as its main anti-tumor components. The poor water solubility, high toxicity, rapid elimination and less selectivity in vivo of bufadienolides limit the application of toad skin. Based on the “unification of drugs and excipients” theory, the toad skin extracts (TSE) and Brucea javanica oil (BJO) nanoemulsions (NEs) were designed to solve the aforementioned problems. BJO as the main oil phase was not only used to prepare the NEs, but played a synergistic therapeutic role combined with TSE. TSE-BJO NEs showed 155 nm particle size, entrapment efficiency of >95% and good stability. TSE-BJO NEs demonstrated superior anti-tumor activity compared with the TSE or BJO NEs alone. The mechanism of TSE-BJO NEs to enhance the antineoplastic efficacy involved several pathways, such as inhibiting cell proliferation, inducing tumor cell apoptosis >40% and arresting cell cycle at G2/M. TSE-BJO NEs could co-deliver drugs into the target cells efficiently and exhibit satisfying synergism. Besides, TSE-BJO NEs facilitated the long circulation of bufadienolides contributing to the high accumulation of drugs at tumor sites and the improvement of anti-tumor efficacy. The study achieves the combinative administration of the toxic TSE and BJO with high efficacy and safety.
研究领域:
药剂学
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肿瘤学
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纳米技术
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DOI
PubMed
高影响因子
Frontiers in Chemistry
·
2021
·
IF 5.221
Li Jianwen,Zhang Zhanxia,Deng Haibin,Zheng Zhan
Cinobufagin is used as a traditional Chinese medicine for cancer therapy. However, it has some disadvantages, such as poor water solubility, short circulating half-life, and low bioavailability. In the present study, a targeted delivery and smart responsive polydopamine (PDA)-based nanomedicine for delivering cinobufagin was rationally designed to improve the anticancer efficacy of the compound for the treatment of lung cancer. The modification of the nanomedicine using folic acid first mediated tumor targeting via the interaction between folic acid and its receptors on tumor cells. After lysosomes escape, the PDA nanomedicine was triggered by the low pH and released its cargo into the tumor microenvironment. The nanomedicine had a better therapeutic effect against lung cancer when used in combination with photothermal therapy. Compared with other nanomedicines used with photothermal therapy, this nanocarrier was not only sensitive to biologically low pH levels for on-demand drug release, but was also biodegradable, breaking down into biocompatible terminal products. Therefore, the proposed drug delivery system with targeted delivery and smart release demonstrated potential as a multifunctional nanoplatform that can enhance the bioavailability and reduce the side effects of chemotherapeutic agents.
研究领域:
药学
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纳米技术
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癌症研究
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生物医学工程
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DOI
PubMed
高影响因子
Metabolomics
·
2023
·Vol. 19
·期 2
·1-13
·
IF 3.6
Zhang Rong,Zhang Zijia,Wu Wenyong,Shi Jingying,Berk Entezar,Li Wei,Deng Yanping,Wang Zhaojun,Hou Jinjun,Long Huali,Lei Min,Wu Wanying
Introduction Lipid metabolism participates in various biological processes such as proliferation, apoptosis, migration, invasion, and maintenance of membrane homeostasis of prostate tumor cells. Bufadienolides, the active ingredients of Chansu, show a robust anti-proliferative effect against prostate cancer cells in vitro, but whether bufadienolides could regulate the lipid metabolism in prostate cancer has not been evaluated. Objectives Our study explored the regulatory effects of bufadienolides on lipid metabolism in human prostate carcinoma cells (PC-3). Methods Untargeted lipidomics and transcriptomics were combined to study the effect of different bufadienolides interventions on lipid and gene changes of PC-3 cells. The key genes related to lipid metabolism and prostate cancer development were verified by qPCR and western blotting. Results Lipidomic analysis showed that the active bufadienolides significantly downregulated the content of long-chain lipids of PC-3 cells. Based on transcriptomic and qPCR analyses, many genes related to lipid metabolism were significantly regulated by active bufadienolides, such as ELOVL6 , CYP2E1 , GAL3ST1 , CERS1 , PLA2G10 , PLD1 , SPTLC3, and GPX2 . Bioinformatics analysis of the Cancer Genome Atlas database and literature retrieval showed that elongation of very long-chain fatty acids protein 6 ( ELOVL6 ) and phospholipase D1 ( PLD1 ) might be important regulatory genes. Western blot analysis revealed that active bufadienolides could downregulate PLD1 protein levels which might promote anti-prostate cancer effect. Conclusions All these findings support that bufadienolides might induce lipid metabolic remodeling by regulating long-chain lipids synthesis and phospholipid hydrolysis to achieve an anti-prostate cancer effect, and PLD1 would probably be the key protein.
研究领域:
肿瘤学
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药理学
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分子生物学
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生物化学
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DOI
PubMed
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
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药理学
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产品关联
DOI
PubMed
高影响因子
MOLECULAR ECOLOGY
·
2024
·
IF 4.5
Juliane P. C. Monteiro,Heike Pröhl,Mariana L. Lyra,Andrés E. Brunetti,Eli C. de Nardin,Thais H. Condez,Célio F. B. Haddad,Ariel Rodríguez
Colour signals play pivotal roles in different communication systems, and the evolution of these characters has been associated with behavioural ecology, integumentary production processes and perceptual mechanisms of the species involved. Here, we present the first insight into the molecular and histological basis of skin colour polymorphism within a miniaturized species of pumpkin toadlet, potentially representing the lowest size threshold for colour polytypism in tetrapods. Brachycephalus actaeus exhibits a coloration ranging from cryptic green to conspicuous orange skin, and our findings suggest that colour morphs differ in their capability to be detected by potential predators. We also found that the distribution and abundance of chromatophores are variable in the different colour morphs. The expression pattern of coloration related genes was predominantly associated with melanin synthesis (including dct , edn1 , mlana , oca2 , pmel , slc24a5 , tyrp1 and wnt9a ). Up-regulation of melanin genes in grey, green and brown skin was associated with higher melanophore abundance than in orange skin, where xanthophores predominate. Our findings provide a significant foundation for comparing and understanding the diverse pathways that contribute to the evolution of pigment production in the skin of amphibians.
研究领域:
进化生物学
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分子生物学
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动物行为学
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两栖动物学
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DOI
PubMed
高影响因子
ENVIRONMENTAL POLLUTION
·
2023
·
IF 8.9
Xinyi Li,Kaiyue Li,Jiayi Li,Zhiqin Zhang,Hongyuan Wang
Endocrine disruptors have been demonstrated to exert adverse effects on growth and development of amphibians by disrupting hormone levels. Tail resorption, which is one of the most remarkable events during amphibian metamorphosis, is closely associated with thyroid hormones levels. However, limited research has been conducted on the effects of endocrine disruptors on tail resorption in amphibians. This study explored the effects of NaClO 4 and T4 on the growth, development and tail resorption during the metamorphosis of Rana Chensinensis. The results demonstrated that exposure to NaClO 4 led to an increase in body size and a delay in metamorphosis of R. Chensinensis tadpoles. Histological analysis revealed that both NaClO 4 and exogenous T4 exposure resulted in thyroid gland injury, and NaClO 4 treatment delayed the degradation of notochord and muscles, thereby delaying tail resorption. Moreover, transcriptome sequencing results showed that apoptosis-related genes ( APAF1 , BAX and CASP6 ) and cell component degradation-related genes ( MMP9 and MMP13 ) were highly expressed in the T4 exposure group, and the expression of oxidative stress-related genes ( SOD and CAT ) was higher in the NaClO 4 exposure group. Taken together, both NaClO 4 and exogenous T4 affect tail resorption in R. Chensinensis , thereby affecting their adaptation to terrestrial life. The present study will not only provide a reference for future experimental research on the effects of other endocrine disruptors on the growth, development and tail resorption of amphibians but will also provide insights into environmental protection and ecological risk assessment.
研究领域:
生态毒理学
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发育生物学
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内分泌学
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分子生物学
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DOI
PubMed
高影响因子
MOLECULAR PHARMACEUTICS
·
2024
·
IF 4.5
Shuai Hong,Changxiu Lin,Junsheng Hu,Jingshu Piao,Ming Guan Piao
Nimodipine is the primary clinical drug used to treat cerebral vasospasm following subarachnoid hemorrhage. Currently, tablets have low bioavailability when taken orally, and injections contain ethanol. Therefore, we investigated a new method of nimodipine administration, namely, nasoencephalic administration. Nasal administration of nimodipine was carried out by attaching the cell-penetrating peptide octa-arginine (R8) to liposomes of nimodipine and incorporating it into a temperature-sensitive in situ gel. The prepared liposomes and gels underwent separate evaluations for in vitro characterization. In vitro release exhibited a significant slow-release effect. In vitro toad maxillary cilia model, RPMI 2650 cytotoxicity, and in vivo SD rat pathological histotoxicity experiments showed that all the dosage from the groups had no significant toxicity to toad maxillary cilia, RPMI 2650 cells, and SD rat tissues and organs, and the cilia continued to oscillate up to 694 ± 10.15 min, with the survival rate of the cells being above 85%. A transwell nasal mucosa cell model and an isolated porcine nasal mucosa model were established, and the results showed that the osmolality of the R8-modified nimodipine liposomal gel to nasal mucosal cells and isolated porcine nasal mucosa was 30.41 ± 2.14 and 65.9 ± 7.34 μg/mL, respectively, which was significantly higher than that of the NM-Solution and PEGylated nimodipine liposome gel groups. Animal fluorescence imaging studies revealed that the R8-modified nimodipine liposomal gel displayed increased brain fluorescence intensity compared to the normal liposomal gel. Pharmacokinetic results showed that after transnasal administration, the AUC(0–∞) of the R8-modified nimodipine liposomal gel was 11.662 ± 1.97 μg·mL–1, which was significantly higher than that of the plain nimodipine liposomal gel (5.499 ± 2.89 μg·mL–1). Brain-targeting experiments showed that the brain-targeting efficiencies of the PEGylated nimodipine liposome gel and R8-modified PEGylated nimodipine liposome gels were 20.44 and 33.45, respectively, suggesting that R8/PEG/Lip-NM-TSG significantly increased the brain-targeting of the drug.
研究领域:
药学
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神经学
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药物递送系统
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药理学
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生物技术
✓ 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）