Ethyl Acetate Fractions of Salvia miltiorrhiza Bunge (Danshen) Crude Extract Modulate Fibrotic Signals to Ameliorate Diabetic Kidney Injury
Abstract
:1. Introduction
2. Results
2.1. Effects of S. miltiorrhiza Extracts on PPAR-α and PPAR-γ Transcriptional Activities in Renal Mesangial Cells Using Dual-Luciferase Reporter Assay
2.2. Effects of S. miltiorrhiza EtOAc Extract on Renal Inflammation/Fibrosis-Related Genes in HG-Stressed Mesangial Cells
2.3. Inhibitory Effects of S. miltiorrhiza Extracts on TGF-β1-Induced Myofibroblast Activation
2.4. In Vivo Effects of EtOAc Extract of S. miltiorrhiza on Renal Gene Expression in STZ-Induced Diabetic Mice
2.5. Renoprotective Effects of S. miltiorrhiza EtOAc Extract STZ-Induced Diabetic Mice
2.6. HPLC Fingerprint Profile of the EtOAc Extract and the Isolation Components
3. Discussion
4. Materials and Methods
4.1. Cell Cultures
4.1.1. In Vitro Model of High Glucose Stress
4.1.2. In Vitro Model of Myofibroblast Activation
4.2. Preparation of the S. miltiorrhiza (Danshen) Extract
4.3. Main Compounds Isolation and HPLC Profile of the EtOAc Extract
4.4. Measurement of PPAR-α and PPAR-γ Activities by Reporter Assay
4.5. RNA Extraction and Quantitative Reverse Transcription-PCR (qRT-PCR)
4.6. Protein Extraction and Western Blot Analysis
4.7. Streptozotocin (STZ)-Induced Diabetic Animal Model and Treatment
4.8. Urine and Blood Biochemistry
4.9. Histological and Immunohistochemical Examinations
4.10. Statistical Analysis
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Genes | Sense (Forward) | Antisense (Reverse) |
---|---|---|
PPAR-γ | 5′-CCAAAGCCTGAGCCCAGA-3′ | 5′-GCACCACTCCCATGGCAT-3′ |
IL-1β | 5′-CCAGGATGAGGACCCAAGCA-3′ | 5′-TCCCGACCATTGCTGTTTCC-3′ |
TGF-β1 | 5′-TGAGTGGCTGTCTTTTGACG-3′ | 5′-TGGGACTGATCCCATTGATT-3′ |
Fibronectin | 5′-CAGCCCCTGATTGGAGTC-3′ | 5′-TGGGTGACACCTGAGTGAAC-3′ |
β-actin | 5′-CGCCAACCGCGAGAAGAT-3′ | 5′-CGTCACCGGAGTCCATCA-3′ |
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Hsu, Y.-C.; Shih, Y.-H.; Ho, C.; Liu, C.-C.; Liaw, C.-C.; Lin, H.-Y.; Lin, C.-L. Ethyl Acetate Fractions of Salvia miltiorrhiza Bunge (Danshen) Crude Extract Modulate Fibrotic Signals to Ameliorate Diabetic Kidney Injury. Int. J. Mol. Sci. 2024, 25, 8986. https://doi.org/10.3390/ijms25168986
Hsu Y-C, Shih Y-H, Ho C, Liu C-C, Liaw C-C, Lin H-Y, Lin C-L. Ethyl Acetate Fractions of Salvia miltiorrhiza Bunge (Danshen) Crude Extract Modulate Fibrotic Signals to Ameliorate Diabetic Kidney Injury. International Journal of Molecular Sciences. 2024; 25(16):8986. https://doi.org/10.3390/ijms25168986
Chicago/Turabian StyleHsu, Yung-Chien, Ya-Hsueh Shih, Cheng Ho, Cheng-Chi Liu, Chia-Ching Liaw, Hui-Yi Lin, and Chun-Liang Lin. 2024. "Ethyl Acetate Fractions of Salvia miltiorrhiza Bunge (Danshen) Crude Extract Modulate Fibrotic Signals to Ameliorate Diabetic Kidney Injury" International Journal of Molecular Sciences 25, no. 16: 8986. https://doi.org/10.3390/ijms25168986