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| Study on the berberine regulates miR-29b to inhibit epithelial-mesenchymal transition of MPC-5 podocytes induced by high glucose |
| Hits 1496 Download times 1170 Received:August 15, 2020 |
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| DOI
10.11656/j.issn.1672-1519.2021.01.25 |
| Key Words
berberine;diabetic nephropathy;podocyte;micro RNA-29b;epithelial-mesenchymal transition |
| Author Name | Affiliation | | GUO Xiaolei | Department of Emergency, First Teaching Hospital of Tianjin University of Traditional Chinese Medicine, Tianjin 300381, China |
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| Abstract
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| [Objective] To investigate the role of miR-29b in the high glucose induced epithelial-mesenchymal transition of MPC-5 podocytes and the protective mechanism of berberine.[Methods] The podocytes of MPC-5 mice were divided into normal group,high glucose group and high glucose + 50 μmol/L berberine group. After 48 hours of culture,TGF-β1 in the supernatant of culture medium was measured by ELISA,the expression of miR-29b was measured by RT-qPCR,and EMT related proteins[α-smooth muscle actin (α-SMA),desmin and podocin] were detected by Western blot. The podocytes were transfected into miR-29b mimic or miR-29b inhibitor respectively,and then cultured with high glucose stimulation and/or berberine. TGF-β1 in the supernatant of the culture solution was determined by ELISA. The expression of EMT-related proteins was detected by Western blot. The double luciferase reporter gene experiment was used to detect whether miR-29b and TGF-β1 can be directly combined.[Results] Compared with the normal group,the expression of miR-29b decreased,the concentration of TGF-β1 increased,the expressions of EMT-related proteins α-SMA and desmin increased,and the expression of podocin decreased in the high glucose group. Compared with high glucose group,the expression of miR-29b in high glucose + berberine group increased,the concentration of TGF-β1,the expression of α-SMA and desmin decreased,and the expression of podocin increased (P<0.01). Compared with the high glucose group,the concentration of TGF-β1 decreased,the expression of α-SMA and desmin decreased and the expression of podocin increased in the high glucose miR-29b mimic group (P<0.01). Compared with high glucose + berberine group,the concentration of TGF-β1,the expression of α-SMA and desmin increased and the expression of podocin decreased in high glucose + berberine miR-29b inhibitor group (P<0.01). The experiment of double luciferase reporter gene showed that miR-29b could directly act on TGF-β1 and inhibit its expression.[Conclusion] Berberine can reduce the expression of TGF-β1 in podocytes induced by high glucose and the degree of epithelial-mesenchymal transition in podocytes by regulating the level of miR-29b. |
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