PHYTOMEDICINE
·
2024
·
IF 6.7
Yu-Qing Li,Xin-Ming Yu,Xue-Mei Shang,Jing-Yi Lin,Rui-Zhi Tan,Jian-Chun Li,Hong-Wei Su,Hong-Ping Shen,Hong-Lian Wang,Li Wang
Background Renal fibrosis is a hallmark of chronic kidney disease (CKD). Smad3 serves as the principal transcription factor mediating the pro-fibrosis effects of TGF-β signaling in renal fibrosis. Biochanin A (BCA), a natural isoflavone, has been shown to attenuate renal fibrosis by inhibiting TGF-β signaling but the detailed mechanisms remain unresolved. This study aimed to elucidate the specific mechanisms by which BCA modulates TGF-β signaling. Methods Renal fibrosis models were established both in vitro , using TGF-β1-stimulated mouse renal tubular TCMK1 cells, and in vivo , employing mice with unilateral ureter obstruction (UUO). RNA-seq was conducted to identify BCA-regulated genes. The AnimalTFDB4.0 database was utilized to predict transcription factors with potential binding to Smad3 promoter. The activities of TGF-β signaling and the cloned mouse Smad3 promoter were assessed using luciferase reporter assays. Plasmid transfection was performed using polyethylenimine in TCMK1 cells or ultrasound microbubbles in UUO kidneys. Gene expression was analyzed by RT-PCR, western blot, and immunohistochemistry assays. Results BCA significantly inhibited TGF-β signaling activity and suppressed TGF-β1-induced fibrotic gene expression in TCMK1 cells. RNA-seq and in silico analyses identified Smad3 as the key gene downregulated by BCA, while leaving Smad2 unaffected. This selective transcriptional suppression of Smad3 by BCA was validated by luciferase reporter assays using the cloned Smad3 promoter. Furthermore, transcription factor binding prediction identified that Klf6, a transcription factor downregulated by BCA, has binding potential to the Smad3 promoter and promotes Smad3 transcription. Klf6 expression was induced in TGF-β1-stimulated TCMK1 cells and UUO kidneys, but this induction was abolished upon BCA treatment. Importantly, Klf6 overexpression restored Smad3 expression and counteracted the anti-fibrosis effects of BCA in both TGF-β1-stimulated TCMK1 cells and UUO kidneys. Conclusion TGF-β-responsive Klf6 transcriptionally transactivates Smad3 expression. BCA exerts anti-renal fibrosis effects by inhibiting the Klf6-Smad3 signaling axis, underscoring its therapeutic potential in the treatment of CKD.
研究领域:
药理学
·
分子生物学
·
肾病学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
SEPARATION AND PURIFICATION TECHNOLOGY
·
2013
·
IF 2.894
Fei-Yue Ma,Cheng-Bo Gu,Chun-Ying Li,Meng Luo,Wei Wang,Yuan-Gang Zu,Ji Li,Yu-Jie Fu
An efficient microwave-assisted aqueous two-phase extraction (MA-ATPE) technique was developed to simultaneously extract and enrich genistein and biochanin A from Dalbergia odorifera T. Chen leaves. Three salts were investigated, and dipotassium hydrogen phosphate was selected to form aqueous two-phase system (ATPS). Box–Behnken Design combined with response surface methodology was applied to optimize the operating parameters of MA-ATPE. The optimal conditions were as follows: salt concentration 20%, absolute alcohol concentration 25%, extraction temperature 45.5 °C, extraction time 10 min and irradiation power 300 W. Under the optimized condition, the extraction yields of genistein and biochanin A reached 1.034 mg/g DM and 2.023 mg/g DM, respectively. Compared with microwave assisted extraction (MAE) and heat reflux extraction (HRE), the contents of genistein and biochanin A in the extracts reached 12.966 mg/g extract and 25.526 mg/g extract, which were 2.48- and 2.30-fold to that of MAE, 2.64- and 2.80-fold to that of HRE. Furthermore, the antioxidant activities of extracts from MA-ATPE, MAE and HRE were 0.342, 0.389, 0.411 mg/mL (IC 50 ), according to the DPPH radical-scavenging assay. Moreover, the phase-forming salt can be recyclable. In conclusion, MA-ATPE was an efficient, eco-friendly and economical technique, which represented an excellent alternative for the extraction of bioactive compounds from plant.
研究领域:
植物化学
·
天然产物提取
✓ 已验证
DOI
高影响因子
Food & Function
·
2021
·Vol. 12
·期 5
·1958-1972
·
IF 5.396
Zhaohui Xue,Xin Gao,Wancong Yu,Qian Zhang,Weichen Song,Shihao Li,Xu Zheng,Xiaohong Kou
Urban particulate matter (UPM), an air pollutant-absorbing toxic substance, can access alveoli, leading to pulmonary diseases. Studies have shown that the water-soluble components of UPM (WS-UPM), containing main toxic substances, can induce oxidative damage in lung cells. In this study, the UPM particle size and composition were detected via instrumental analysis. The isoflavones (biochanin A (BCA), formononetin and daidzein) from chickpeas possess biological antioxidant properties. The present study aimed to investigate the mechanism of the oxidative damage induced by WS-UPM, and the protective role of isoflavones in human alveolar basal epithelial cells. The antioxidant activity of BCA, formononetin and daidzein was investigated through the total reduction capacity, diphenylpicrylhydrazine radical (DPPH), superoxide radical, and hydroxyl radical scavenging capacity detection. We also established cell models in vitro to further explore the BCA-protective mechanism. BCA presented a significant protection, and increased the levels of antioxidant makers including superoxide dismutase (SOD), catalase (CAT), and glutathione (GSH). The effects were also reflected as the reduction of malondialdehyde (MDA) and nitric oxide (NO). Moreover, results obtained from RT-PCR and western blot techniques revealed that MEK5/ERK5 played an indispensable role in regulating the antioxidant effect of BCA, alleviating WS-UPM-induced lung injury. Furthermore, BCA mitigated WS-UPM-exposed damage through upregulating the Nrf2 signaling pathway to enhance the antioxidase expression downstream of Nrf2. In summary, our findings indicated that the WS-UPM-induced pulmonary disease was involved in oxidative stress and the MEK5/ERK5-Nrf2 signaling pathway, and BCA regulated the WS-UPM-induced lung damage via upregulation of the MEK5/ERK5-Nrf2 pathway.
研究领域:
环境科学
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毒理学
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药理学
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细胞生物学
✓ 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）