Genes
·
2021
·Vol. 12
·期 8
·1287
·
IF 4.096
Shouxiang Sun,Yumei Wang,Pei-Tian Goh,Mónica Lopes-Marques,L. Filipe C. Castro,Óscar Monroig,Meng-Kiat Kuah,Xiaojuan Cao,Alexander Chong Shu-Chien,Jian Gao
Elongation of very long-chain fatty acid (Elovl) proteins are key enzymes that catalyze the rate-limiting step in the fatty acid elongation pathway. The most recently discovered member of the Elovl family, Elovl8, has been proposed to be a fish-specific elongase with two gene paralogs described in teleosts. However, the biological functions of Elovl8 are still to be elucidated. In this study, we showed that in contrast to previous findings,elovl8is not unique to teleosts, but displays a rather unique and ample phylogenetic distribution. For functional determination, we generatedelovl8a(elovl8a−/−) andelovl8b(elovl8b−/−) zebrafish using CRISPR/Cas9 technology. Fatty acid composition in vivo and zebrafish liver cell experiments suggest that the substrate preference of Elovl8 overlapped with other existing Elovl enzymes. Zebrafish Elovl8a could elongate the polyunsaturated fatty acids (PUFAs) C18:2n-6 and C18:3n-3 to C20:2n-6 and C20:3n-3, respectively. Along with PUFA, zebrafish Elovl8b also showed the capacity to elongate C18:0 and C20:1. Gene expression quantification suggests that Elovl8a and Elovl8b may play a potentially important role in fatty acid biosynthesis. Overall, our results provide novel insights into the function of Elovl8a and Elovl8b, representing additional fatty acid elongases not previously described in chordates.Keywords:elovl8;gene knockout;zebrafish;fatty acid synthesis
研究领域:
分子生物学
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遗传学
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生物化学
✓ 已验证
DOI
PubMed
高影响因子
JOURNAL OF ETHNOPHARMACOLOGY
·
2023
·
IF 5.4
Xueping Wei,Junsong Jing,Rongrong Huang,Ting Zhou,Lianhao Wu,Guoteng Ou,Youping Wu,Jingjin Hu,Wenwen Zhu,Yueguo Wu,Yuanyuan Li,Sheng Zhang,Zhenqiang You
Ethnopharmacological relevance Pulmonary fibrosis (PF) is an irreversible lung disease that severely affects human respiratory function. Traditionally, the natural plant Quzhou Fructus Arantii (QFA) has therapeutic effects on respiratory diseases. However, the effects and the mechanism of anti-fibrotic have not been elucidated. Aim of the study In this study, QFAE-nB was extracted from QFA, the aims of this study include understanding the correlation between Bleomycin (BLM)-induced PF and STING pathway in mice, as well as exploring the role and mechanisms of QFAE-nB in the treatment of PF. Materials and methods QFAE-nB was extracted from QFA, six main chemical components in QFAE-nB were identified by HPLC–QTOF–MS/MS, and quantitative analysis was conducted by HPLC. qPCR and Western blot were used to verify the molecular mechanism of QFAE-nB, and the anti-fibrotic effect of QFAE-nB was determined by hematoxylin-eosin (HE) staining and Masson staining as well as immunohistochemistry. TREX1-KO and STING-KO mice were used to verify the relationship between STING and PF and the important target action of QFAE-nB. Results Six main flavonoids in QFAE-nB were identified as eriocitrin (0.76%), neoeriocitrin (2.79%), narirutin (4.31%), naringin (35.41%), hesperidin (1.74%), and neohesperidin (27.18%). The results showed that BLM-induced PF was associated with its exacerbated release of proinflammatory factors and chemokines in lung tissues. In addition, QFAE-nB alleviated BLM-induced lung fibrosis in mice by inhibiting the activation of the STING signaling pathway and reducing the signal transduction of TBK1–IRF3 and TBK1–NF-κB pathways. Notably, knockout of the TREX1 gene caused massive inflammation and even induced PF in the lung tissues, whereas QFAE-nB effectively alleviated inflammation and reduced PF. The deletion of the STING gene suppressed BLM-induced PF and inflammation, but STING-KO mice treated with QFAE-nB showed even lower expression levels of proinflammatory factors and chemokine. Conclusions The STING pathway plays an important role in PF, and QFAE-nB alleviates PF by mainly targeting the inhibition of the STING pathway to reduce inflammation. Together, the study paves the way for targeting the STING pathway in PF treatment.
研究领域:
药理学
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民族药理学
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分子生物学
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免疫学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Molecular Medicine Reports
·
2025
·Vol. 32
·期 1
·1-15
·
IF 3.5
Zhenfei Mo,Chengcheng Su,Jinxia Liu,Jiabo Ren,Lu Liu,Yueming Wang,Yanqin Li,Chunsun Li,Zhen Yang,Xiuqing Ma,Liangan Chen
The oncogene lung cancer metastasis‑related protein 1 (LCMR1) is associated with neoplastic diseases and LCMR1 conditional knockout affects cell homeostasis. In the present study, the role of LCMR1 in lipopolysaccharide (LPS)‑induced acute lung injury (ALI) was investigated. Firstly, wild‑type C57BL/6 mice were used to establish an LPS‑induced ALI model via intratracheal injection of LPS, and the expression of LCMR1 was examined at 24, 48, 72 and 96 h after injury. The LPS‑induced lung injury model was subsequently constructed in mice with conditional knockout of LCMR1 in type II alveolar epithelial cells (AEC‑II). Subsequently, histopathological analysis, lung wet/dry weight ratio comparisons and lung function tests were performed; survival rates after LPS challenge of the conditional knockout mice were measured; bronchoalveolar lavage fluid (BALF) was collected, and the concentrations of protein and inflammatory cytokines in BALF were measured; and transmission electron microscopy of lung tissue was conducted to evaluate the degree of lung injury. To further investigate the mechanism, a lung‑on‑a‑chip model with overexpression or knockdown of LCMR1 was constructed to simulate the alveolar environment under LPS treatment. The expression levels of E‑cadherin and pro‑pulmonary surfactant C precursor (proSP‑C) in the chips were determined by immunofluorescence, and the integrity of the air‑blood barrier was analyzed using a permeability assay. In the mouse model, LCMR1 expression was downregulated in wild‑type mice with LPS‑induced lung injury. LCMR1 conditional knockout in AEC‑II caused increased mortality, impaired lung function, aggravated pathological damage and increased the inflammatory response in mice with LPS‑induced ALI. Furthermore, in the lung‑on‑a‑chip model, LCMR1 knockdown reduced the expression of E‑cadherin and proSP‑C, and impaired the air‑blood barrier function, whereas LCMR1 overexpression attenuated these effects, which may be related to cell differentiation dysfunction and enhanced apoptosis. In conclusion, the present study revealed that LCMR1 deficiency may exacerbate LPS‑induced ALI and could be considered a novel target for intervention in ALI.
研究领域:
医学
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分子生物学
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病理学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
RSC Advances
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2025
·Vol. 15
·期 30
·24437
·
IF 4.6
Feng Wen,Yongjun Wu,Jing Jin,Lincheng Zhang,Shuoqiu Tong,Cen Li,Li Ran,Yuzhang Zhu
Fermented soybean products are an integral part of the Chinese diet, valued for their distinctive aroma. Bacillus subtilis ( B. subtilis ) BJ3-2 is a highly effective fermentation strain widely used in the industrial production of bacterial-type douchi. However, the specific aroma characteristics of soybeans fermented by this strain remain unclear. This study utilized gene knockout, GC-O-MS, and multivariate statistical analysis to comprehensively investigate the aroma components of soybeans fermented by B. subtilis BJ3-2. For the first time, 12 key aroma skeleton compounds were identified, including guaiacol, 2,5-dimethylpyrazine, and 2-methylbutyric acid, which primarily contribute to the smoky, roasted, and sweaty aroma attributes of fermented soybeans. Additionally, 17 compounds were considered to have important coordinating effects on soybean aroma, such as 3-hydroxy-2-butanone (buttery, creamy). Partial least squares regression (PLSR) analysis demonstrated a good correlation between key aroma compounds and sensory attributes. Gene knockout of fadA and bkdB resulted in a significant increase in 2,5-dimethylpyrazine and 1-octen-3-ol levels, along with a marked reduction in 2-methylbutyric acid levels. These changes enhanced the roasted and beany notes, reduced the sweaty aroma, and ultimately improved the overall aroma profile. These findings provide insights into the genetic regulation of aroma in fermented soybean products and form a foundation for reducing undesirable aromas and improving the overall aroma quality of industrially produced douchi. Bacillus subtilis fermented soy food is deeply loved by consumers, and its unique aroma characteristics are influenced by the fadA and bkdB genes.
研究领域:
食品科学
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微生物学
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分子生物学
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感官分析
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生物技术
✓ 已验证
产品关联
DOI
PubMed
高影响因子
XENOBIOTICA
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2019
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IF 1.902
Huanying Ye,Danjuan Sui,Wei Liu,Yuannan Yuan,Zhen Ouyang,Yuan Wei
1. CYP2C11 is the most abundant isoform of cytochrome P450s (CYPs) in male rats and is considered the main enzyme for warfarin metabolism.2. To further access the in vivo function of CYP2C11 in warfarin metabolism and efficacy, a CYP2C11-null rat model was used to study warfarin metabolism with both in vitro and in vivo approaches. Prothrombin time (PT) of warfarin was also determined.3. The maximum rate of metabolism (Vmax) and intrinsic clearance (CLint) of liver microsomes from CYP2C11-null males were reduced by 37 and 64%, respectively, compared to those in Sprague Dawley (S-D) rats. The Km of liver microsomes from CYP2C11-null males was increased by 73% compared to that of S-D rats. The time to reach the maximum plasma concentration (Tmax) of warfarin in CYP2C11-null males was significantly delayed compared to that in S-D males, and the CL rate was also reduced. The PT of CYP2C11-null rats was moderately longer than that of S-D rats.4. In conclusion, the clearance rate of warfarin was mildly decreased and its anticoagulant effect was moderately increased in male rats following CYP2C11 gene knockout. CYP2C11 played a certain role in the clearance and efficacy of warfarin, while it did not seem to be essential.
研究领域:
药理学
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分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
·
2025
·
IF 2.2
Youming Shi,Wenjiao Gu,Yujie Wei,Shuangyi Li,Shining Zhang,Ying jiang,Chuanjian Chen,Ting Liu,Limei Shuai,Xiaochun Zhou,Futian Tang
Purpose Atherosclerosis (AS) and atherosclerotic calcification (AC) are closely related to the cardiovascular diseases, largely due to its induction of vasomotor dysfunction. We previously reported that Calpain-1 inhibitor attenuated AS and AC. The present study was designed to investigate the effects and potential mechanisms of Calpain-1 knockout (Calpain-1 KO) in Apolipoprotein E KO (ApoE KO) mice on AS, AC, aortic vasomotor function as well as the liver dysfunction. Methods We hybridized ApoE KO mice with Calpain-1 KO mice to obtain ApoE /Calpain-1 double KO (A×C DKO) mice. The formation of AS and AC was evaluated and liver function was determined. Aortic vasomotor function was assessed. Contents of TNF-α, IL-6, IL-18, IL-1β and NO and the activity of AST, ALT, ALP and eNOS in serum were quantified. The mRNA expression of CD68, SR-A, CD36, PPAR-γ, LXR-α, ABCA1, BMP-2, OPN, ALP and Runx2 in aorta and/or liver were measured. Results The results showed that in comparison to C57 mice, ApoE KO mice demonstrated the significant increases in the areas of AS and AC, the increases in the mRNA expression of CD68 in the aorta, the increases in the AST, ALT and ALP activity in serum. ApoE KO mice also showed the dysfunction of the aortic contraction induced by phenylephrine and of the relaxation induced by acetylcholine. However, compared with ApoE KO mice, A×C DKO mice exhibited the significant attenuation of AS and AC and the downregulation of mRNA expression of CD68 in aorta. A×C DKO mice revealed the reduction of AST, ALT and ALP activity in serum, the improvements in aortic contraction and relaxation as well as the increases in eNOS activity and NO content in serum. A×C DKO mice also showed the decreases in the contents of TNF-α, IL-6, IL-18 and IL-1β in serum. The mRNA expression of CD68 in aorta, SR-A and CD36 in both aorta and liver of A×C DKO mice was downregulated, while that of PPAR-γ, LXR-α, and ABCA1 was upregulated in comparison of ApoE KO mice. In addition, the mRNA expression of BMP-2, OPN, ALP and Runx2 in aorta of A×C DKO mice was downregulated in comparison of ApoE KO mice. Conclusion The results suggested that deficiency of Calpain-1 attenuated the formation of AS and AC and improved vasomotor and liver dysfunction in ApoE KO mice through anti-inflammation, and modulation of the mRNA expression of genes related to AS and AC.
研究领域:
心血管疾病研究
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分子生物学
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药理学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Cellular & Molecular Immunology
·
2024
·
IF 21.8
Wu Xiaoxia,Zhao Xiaoming,Li Fang,Wang Yang,Ou Yangjing,Zhang Haiwei,Li Xiaoming,Wu Xuanhui,Wang Lingxia,Li Ming,Zhang Yue,Liu Jianling,Xing Mingyan,Liu Han,Tan Yongchang,Wang Yangyang,Xie Yangyang,Zhang Hanwen,Luo Yan,Li Hong,Wang Jing,Sun Liming,Li Yu,Zhang Haibing
The hypersecretion of cytokines triggers life-threatening systemic inflammatory response syndrome (SIRS), leading to multiple organ dysfunction syndrome (MODS) and mortality. Although both coagulopathy and necroptosis have been identified as important factors in the pathogenesis of SIRS, the specific cell types that undergo necroptosis and the interrelationships between coagulopathy and necroptosis remain unclear. In this study, we utilized visualization analysis via intravital microscopy to demonstrate that both anticoagulant heparin and nonanticoagulant heparin (NAH) pretreatment protect mice against TNF-α-induced mortality in SIRS. Moreover, the deletion of Mlkl or Ripk3 resulted in decreased coagulation and reduced mortality in TNF-α-induced SIRS. These findings suggest that necroptosis plays a key role upstream of coagulation in SIRS-related mortality. Furthermore, using a genetic lineage tracing mouse model (Tie2-Cre;Rosa26-tdT ), we tracked endothelial cells (ECs) and verified that EC necroptosis is responsible for the vascular damage observed in TNF-α-treated mice. Importantly, Mlkl deletion in vascular ECs in mice had a similar protective effect against lethal SIRS by blocking EC necroptosis to protect the integrity of the endothelium. Collectively, our findings demonstrated that RIPK3–MLKL-dependent necroptosis disrupted vascular integrity, resulting in coagulopathy and multiorgan failure, eventually leading to mortality in SIRS patients. These results highlight the importance of targeting vascular EC necroptosis for the development of effective treatments for SIRS patients.
研究领域:
免疫学
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病理生理学
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细胞生物学
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炎症
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血管生物学
✓ 已验证
DOI
PubMed
高影响因子
Frontiers in Cell and Developmental Biology
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2021
·
IF 6.684
Salehpour Aria,Rezaei Mohammad,Khoradmehr Arezoo,Tahamtani Yaser,Tamadon Amin
Despite extensive studies on type 2 diabetes mellitus (T2DM), there is no definitive cure, drug, or prevention. Therefore, for developing new therapeutics, proper study models of T2DM is necessary to conduct further preclinical researches. Diabetes has been induced in animals using chemical, genetic, hormonal, antibody, viral, and surgical methods or a combination of them. Beside different approaches of diabetes induction, different animal species have been suggested. Although more than 85% of articles have proposed rat (genus Rattus) as the proper model for diabetes induction, zebrafish (Danio rerio) models of diabetes are being used more frequently in diabetes related studies. In this systematic review, we compare different aspects of available methods of inducing hyperglycemia referred as T2DM in zebrafish by utilizing a scoring system. Evaluating 26 approved models of T2DM in zebrafish, this scoring system may help researchers to compare different T2DM zebrafish models and select the best one regarding their own research theme. Eventually, glyoxalase1 (glo1−/−) knockout model of hyperglycemia achieved the highest score. In addition to assessment of hyperglycemic induction methods in zebrafish, eight most commonly proposed diabetic induction approval methods are suggested to help researchers confirm their subsequent proposed models.
研究领域:
医学
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生物学
✓ 已验证
DOI
PubMed
高影响因子
Parasites & Vectors
·
2024
·Vol. 17
·期 1
·1-10
·
IF 3
Huang Yue,Li Jinli,Pei Shifeng,You Heng,Liu Huimin,Guo Yaqiong,Xu Rui,Li Na,Feng Yaoyu,Xiao Lihua
Background The dimerizable Cre recombinase system (DiCre) exhibits increased leaky activity in Cryptosporidium, leading to unintended gene editing in the absence of induction. Therefore, optimization of the current DiCre technique is necessary for functional studies of essential Cryptosporidium genes.Methods Based on the results of transcriptomic analysis of Cryptosporidium parvum stages, seven promoters with different transcriptional capabilities were screened to drive the expression of Cre fragments (FKBP-Cre59 and FRB-Cre60). Transient transfection was performed to assess the effect of promoter strength on leakage activity. In vitro and in vivo experiments were performed to evaluate the leaky activity and cleavage efficiency of the optimized DiCre system by polymerase chain reaction (PCR), nanoluciferase, and fluorescence analyses.Results The use of promoters with lower transcriptional activity, such as pcgd6_4110 and pcgd3_260, as opposed to strong promoters such as pActin, pα-Tubulin, and pEnolase, reduced the leakage rate of the system from 35–75% to nearly undetectable levels, as verified by transient transfection. Subsequent in vitro and in vivo experiments using stable lines further demonstrated that the optimized DiCre system had no detectable leaky activity. The system achieved 71% cleavage efficiency in vitro. In mice, a single dose of the inducer resulted in a 10% conditional gene knockout and fluorescent protein expression in oocysts. These fluorescently tagged transgenic oocysts could be enriched by flow sorting for further infection studies.Conclusion sA DiCre conditional gene knockout system for Cryptosporidium with good cleavage efficiency and reduced leaky activity has been successfully established.Graphical Abstract
研究领域:
寄生虫学
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分子生物学
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基因组编辑
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Systems Microbiology and Biomanufacturing
·
2024
·
IF 0
Huang Yu,Li Hedan,Zhao Guihong,Hu Xiaoqing,Wang Xiaoyuan
The production of L-alanine was enhanced in Corynebacterium glutamicum ATCC13869 through metabolic engineering of the biosynthesis pathways of L-alanine and fatty acids. Strains Δ fasB , Δ fasBR , Δ fasB Δ pks13 and Δ fasBR Δ pks13 were constructed and exhibited increased L-alanine yields up to 17.29 g/L. Different from Δ fasB mutant constructed from C. glutamicum ATCC13032 in which L-glutamate production accumulated, the muatnt Δ fasB constructed from C. glutamicum ATCC13869 in this study significantly produce L-alanine without L-glutamate accumulation. Transcriptional level analysis revealed that the knockout of fasB upregulated the expression levels of the genes related to L-alanine synthesis but downregulated those associated with fatty acid synthesis, confirming the redirection of metabolic flux from fatty acid synthesis to L-alanine synthesis in these strains. L-alanine productions were further enhanced in strains Δ fasB and Δ fasBR through the combinatorial expression of heterologous genes Bacillus subtilis alaD encoding alanine dehydrogenase and Escherichia coli alaE encoding alanine export protein, and the yields reached 55.21 g/L and 54.95 g/L, respectively. Finally, 69.9 g/L L-alanine was obtained in Δ fasB /pJYW-5- alaDE after 60 h of fermentation by supplementing glucose. Our data indicate that disrupting the fatty acid biosynthesis could redirect metabolic flux towards L-alanine biosynthesis. These results provide a new strategy for increasing the production of L-alanine in C. glutamicum . Graphical Abstract
研究领域:
微生物学
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生物工程
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分子生物学
✓ 已验证
DOI
GENE
·
2023
·
IF 3.5
Shaolin Xie,Yongyong Feng,Aiguo Zhou,Zhiyuan Lu,JixingZou
Cytochrome P450 1 (CYP1) is an important enzyme family involved in the metabolism of pollutants, and used as a biomarker to monitor environmental pollution. In this study, a fluorescence-labeled cyp1a zebrafish line, named as KI ( cyp1a +/+ -T2A-mCherry) (KICM), was originally constructed to monitor dioxin-like compounds in the environment. However, the cyp1a gene expression in the KICM line was inhibited by the fluorescence labeling, thus leading to a significantly increased sensitivity of KICM zebrafish line to PAHs. Then, a cyp1a knockout zebrafish line, named KOC, were constructed for comparative analysis with the cyp1a low-expression line. Interestingly, knockout of the cyp1a gene did not increase the sensitivity of zebrafish to PAHs as significantly as the cyp1a low-expression line. So, the expression levels of related genes in the aryl hydrocarbon receptor pathway were analyzed and the results showed that the expression level of cyp1b in KOC group was significantly higher than that of wild type and KICM under the same PAH exposure. This indicated that the effect of losing cyp1a was compensated by inducing expression of cyp1b . In conclusion, two new zebrafish models including cyp1a low-expression line and cyp1a knockout line were constructed in this study, which may provide a convenient model for subsequent studies on the toxicity mechanism of PAHs and the role of cyp1a in detoxification.
研究领域:
环境毒理学
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分子生物学
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遗传学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Foods
·
2023
·Vol. 12
·期 16
·2987
·
IF 5.2
Cheng Guo,Wangyang Shen,Weiping Jin,Xiwu Jia,Zhili Ji,Jinling Li,Bin Li
The composition of green tea cream is extremely complex, and identification of key components is a prerequisite for elucidating its microstructure formation mechanism. This study examined the dynamic changes in the content of components and properties of colloid particles during the formation process of tea cream by chemical analysis and dynamic laser scattering (DLS). A “knock-out/knock-in” method was developed and used to further explore the relationship between the interaction of these components and the microstructure formation of tea cream. The results revealed that polysaccharides, proteins, epigallocatechin gallate (EGCG), and caffeine were the main components involved in tea cream formation. These components participated in the formation process in the form of polysaccharide–protein and EGCG–caffeine colloidal particles. Consequently, there were synchronized dynamic changes in the levels of polysaccharides, proteins, EGCG, and caffeine. The “knock-out/knock-in” experiment revealed that the interactions between EGCG or caffeine and macro-molecule components were not the key factors in tea cream microstructure formation. However, it was found that the complexation between EGCG and caffeine played a crucial role in the formation of tea cream. The findings suggested that decreasing the concentrations of EGCG and caffeine could be useful in controlling tea cream formation during tea beverage processing and storage.Keywords:green tea cream;dynamic formation process;key components;colloid particles;“knock-out/knock-in” method
研究领域:
食品科学
·
化学
✓ 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）