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Outline
Diabetes mellitus is mainly caused by insufficient insulin secretion or impaired insulin action, and the patients exhibit varying degrees of pancreatic islet β-cell dysfunction. In the present study, Rin-m5F pancreatic islet β-cells were used to investigate the protective effect of puerarin against islet β-cell injury and its underlying mechanism. The Cell Counting Kit-8 (CCK-8) assay was performed to evaluate cell viability, so as to screen the optimal concentrations of glucose (as the injury inducer) and puerarin (as the therapeutic agent). Flow cytometry was employed to evaluate the effect of puerarin on apoptosis in the glucose-induced islet β-cell injury model. Molecular docking technology was used to analyze the binding energy between puerarin and B-cell lymphoma-2 (Bcl-2) protein, and Western blot analysis was conducted to analyze the expression levels of Bcl-2 and its downstream-related proteins. The results of CCK-8 showed that 50 mmol·L-1 glucose induced significant injury to Rin-m5F pancreatic islet β-cells, which was improved by puerarin (25-200 μmol·L-1) in a dose-dependent manner. Flow cytometry results demonstrated that puerarin effectively reduced cell apoptosis. Molecular docking analysis revealed a favorable binding energy between puerarin and Bcl-2 protein. Further, Western blot results indicated that puerarin upregulated the expression of Bcl-2, while downregulated the expression of apoptotic pathway-related proteins, including Bcl-2-associated X protein (Bax), cysteinyl aspartate specific proteinase-3 (Caspase-3), and poly (ADP-ribose) polymerase 1 (PARP-1). Additionally, puerarin promoted the expression of microtubule-associated protein 1 light chain 3 (LC3) Ⅱ/Ⅰ. After co-treated with Venetoclax, a Bcl-2 inhibitor, puerarin failed to protect against islet β-cell injury, inhibit apoptosis, and upregulate the expression of Bcl-2. These findings suggest that the protective effect of puerarin on islet β-cell injury is abrogated when Bcl-2 is inhibited. In conclusion, puerarin alleviates pancreatic islet β-cell injury by activating Bcl-2, providing a basis for the further investigation of the mechanism underlying the therapeutic effect of traditional Chinese medicine prescriptions containing Pueraria lobata in the treatment of diabetes mellitus.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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