Qingda Granule Attenuates Angiotensin II-Induced Renal Apoptosis and Activation of the p53 Pathway

Figures

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  FIGURE 1. QDG attenuates Ang II-induced elevation of blood pressure and renal injury. (A) SBP, (B) DBP, and (C) MAP of mice from each group were monitored prior to the experiment and once a week for 4 weeks by using the CODA™ noninvasive blood pressure system. (D) HE staining was performed to determine the pathological changes of renal tissues from each group. The representative images were taken at a magnification of ×400 (scale bar 60 μm). n = 5 for each group. Data were presented as mean ± SD; *p < 0.05 vs. the Control group, #p < 0.05 vs. the Ang II group.

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  FIGURE 2. QDG treatment on gene expression profiling of renal tissues in Ang II-infused mice. RNA-seq was performed to determine the DETs in renal tissues from each group. (A) Hierarchical clustering plots and (B) volcano plots were used to compare gene expression profiles (|fold change| ≥ 2, p < 0.05). (C) Integrative analysis was performed to identify the integrated transcripts between the two comparisons. The overlapped area in the left panel represents decreased transcript number (261) in the Ang II group but increased in the Ang II + QDG group. The overlapped area in the right panel represents increased transcript number (401) in the Ang II group but decreased in the Ang II + QDG group.

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  FIGURE 3. GO enrichment analysis. GO enrichment analysis was performed based on the DETs from both comparisons of (A) Ang II vs. Control and (B) Ang II + QDG vs. Ang II. The top 30 enriched items of the cellular composition (left panel), biological processes (middle panel), and molecular function (right panel).

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  FIGURE 4. KEGG pathway enrichment analysis. (A) KEGG enrichment analysis was performed to identify the enriched signaling pathway in both comparisons of Ang II vs. Control (left panel) and Ang II + QDG vs. Ang II (right panel). The top 30 enriched signaling pathways were presented. (B) Integrative analysis was performed to identify the integrated transcripts between the two comparisons. The overlapped area represents the number of enriched signaling pathways in both comparisons of Ang II vs. Control and Ang II + QDG vs. Ang II.

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  FIGURE 5. QDG attenuates Ang II-induced renal cell apoptosis in vivo and in vitro. The TUNEL assay was performed to observe cell apoptosis in renal tissues of mice from each group (A) and the percentage of TUNEL-positive cells (B). Viability of NRK-52E cells after treatment with QDG, as analyzed by the CCK-8 assay (C). The TUNEL assay was performed to observe cell apoptosis in NRK-52E cells from each group (E) and the percentage of TUNEL-positive cells (D). Representative dot plot of Annexin V- and PI-stained cells (F) and a columnar graph that comprised the percentage of apoptosis in NRK-52E cells (G). Data were presented as mean ± SD; *p < 0.05 vs. the Control group, #p < 0.05 vs. the Ang II group.

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  FIGURE 6. QDG inhibits activation of p53 signaling pathways induced by angiotensin (Ang II). Western blot analysis was performed to determine the protein expression of p53, BAX, BCL2, and cle-caspase-9/caspase-3. GAPDH was used as the internal control. Data were presented as mean ± SD; *p < 0.05 vs. the Control group, #p < 0.05 vs. the Ang II group.