Tanshinone IIA down-regulates -transforming growth factor beta 1 to relieve renal tubular epithelial cell inflammation and pyroptosis caused by high glucose

Figures

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  Figure 1.. Prediction of tanshinone IIA as the effective component of salvia miltiorrhiza in DN and its structural formula. (a) The intersected targets of salvia miltiorrhiza/tanshinone IIA and DN. (b) pubChem analysis of the chemical structure of tanshinone IIA.

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  Figure 2.. Prediction and verification of the effect of tanshinone IIA on TGFB1. (a) String database predicted protein interactions. (b) Western blotting was used to detect the effect of tanshinone IIA on the expression of TGFB1 protein. **P< 0.01, ***P< 0.005.

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  Figure 3.. High glucose did not cause the apoptosis of renal tubular epithelial cells. (a) The levels of TNF-α, IL-6 were detected by RT-qPCR. (b) Flow cytometry was used to detect the effect of high glucose on HK-2 cell apoptosis. (c) Western blotting was used to detect the expression levels of apoptosis-related proteins in each group. **P< 0.01, ***P< 0.005.

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  Figure 4.. High glucose induced renal tubular epithelial cell damage by the pyroptosis pathway. (a) The levels of IL-1β and IL-18 were detected by RT-qPCR. (b) Western blotting was used to detect matured IL-1β expression level in each group. (c) Western blotting was used to detect the expression levels of caspase-1 and N-GSDMD proteins in each group. (d) The pyroptosis of renal tubular epithelial cells was detected by N-GSDMD and TUNEL immunofluorescence double staining. **P < 0.01, ***P< 0.005.

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  Figure 5.. The role of tanshinone IIA in hyperglycemia-induced HK-2 cell injury and its relationship with apoptosis pathway. High glucose-cultured HK-2 cells were treated with tanshinone IIA (1, 5, 10 μM) and tolbutamide. (a) The levels of TNF-α, IL-6 were detected by RT-qPCR. (b) Flow cytometry was used to detect the effect of tanshinone IIA on HK-2 cell apoptosis. (c) The protein expression of genes-related to cell apoptosis was detected using western blot. *P < 0.05, **P < 0.01, ***P< 0.005.

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  Figure 6.. The role of tanshinone IIA in hyperglycemia-induced cell injury and its relationship with pyroptosis pathway. (a) The levels of IL-1β and IL-18 were detected by RT-qPCR. (b) Western blotting was used to detect matured IL-1β expression level. (c) Western blotting was used to detect the expression levels of caspase-1 and N-GSDMD proteins in each group. (d) The effect of tanshinone IIA on pyroptosis of HK-2 cells was detected by N-GSDMD and TUNEL immunofluorescence double staining. *P < 0.05, **P< 0.01, ***P< 0.005.

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  Figure 7.. The role of TGFB1 in the in-vitro model of DN after tanshinone IIA treatment. (a-b) The levels of IL-1β, IL-18, TNF-α, IL-6 were detected by RT-qPCR. (c) Flow cytometry was used to detect the effect of TGFB1 on HK-2 cell apoptosis. (d) The effect of TGFB1 on pyroptosis of HK-2 cells was detected by N-GSDMD and TUNEL immunofluorescence double staining. (e) Western blotting was used to detect the expression levels of caspase-3 and caspase-9, caspase-1 and N-GSDMD proteins in each group. (f) The levels of FN and COL-1 were detected by RT-qPCR. (g) The levels of p65, p-IKKβ, p-IκBα in HK-2 cells under indicated transfections. **P< 0.01, ***P< 0.005, compared with HG group; ##P < 0.01, ###P < 0.005, compared with HG+tanshinone IIA+pcDNA-NC group; &P < 0.05 compared with the HG+tanshinone IIA+pcDNA-NC group.