Korean J Physiol Pharmacol.  2021 Sep;25(5):413-423. 10.4196/kjpp.2021.25.5.413.

Curcumin attenuates renal ischemia reperfusion injury via JNK pathway with the involvement of p300/CBP-mediated histone acetylation

• Yang L
• Chen X
• Bi Z
• Liao J
• Zhao W
• Huang W

• KMID: 2519404
• DOI: http://doi.org/10.4196/kjpp.2021.25.5.413

Abstract

Apoptosis is proved responsible for renal damage during ischemia/reperfusion. The regulation for renal apoptosis induced by ischemia/reperfusion injury (IRI) has still been unclearly characterized to date. In the present study, we investigated the regulation of histone acetylation on IRI-induced renal apoptosis and the molecular mechanisms in rats with the application of curcumin possessing a variety of biological activities involving inhibition of apoptosis. Sprague–Dawley rats were randomized into four experimental groups (SHAM, IRI, curcumin, SP600125). Results showed that curcumin significantly decreased renal apoptosis and caspase-3/-9 expression and enhanced renal function in IRI rats. Treatment with curcumin in IRI rats also led to the decrease in expression of p300/cyclic AMP response element-binding protein (CBP) and activity of histone acetyltransferases (HATs). Reduced histone H3 lysine 9 (H3K9) acetylation was found near the promoter region of caspase-3/-9 after curcumin treatment. In a similar way, SP600125, an inhibitor of c-Jun N-terminal kinase (JNK), also attenuated renal apoptosis and enhanced renal function in IRI rats. In addition, SP600125 suppressed the binding level of p300/CBP and H3K9 acetylation near the promoter region of caspase-3/-9, and curcumin could inhibit JNK phosphorylation like SP600125. These results indicate that curcumin could attenuate renal IRI via JNK/p300/CBP-mediated anti-apoptosis signaling.

Keyword

Curcumin; Histone acetylation; MAP kinase signaling system; p300/CBP; Reperfusion Injury for Renal IRI

Figure

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  Fig. 1 Serum creatinine (Scr) and blood urea nitrogen (BUN) levels were used for assessing renal function. Scr (A) and BUN (B) levels in SHAM group, IRI group and IRI/Cur group. Statistical significance was determined by one-way ANOVA followed by Fisher’s Least Significant Difference test. n = 8 per group. IRI, ischemia/reperfusion injury; Cur, curcumin. *p < 0.05 vs. SHAM group; #p < 0.05 vs. IRI group.

• 

  Fig. 2 Histopathological investigation in the presence or absence of curcumin treatment. (A) Representative images of renal tissue sections stained with H&E and PAS. Black arrows indicate proximal tubule injuries mainly including brush border loss and flatten cells. (B) Semi-quantitative scoring of renal injuries based on morphological changes in renal tissue sections stained with H&E and PAS. n = 8 per group. IRI, ischemia/reperfusion injury; Cur, curcumin. *p < 0.05 vs. SHAM group; #p < 0.05 vs. IRI group.

• 

  Fig. 3 TUNEL assay was used for in situ detection of apoptosis in renal tissue sections. (A) Representative images of renal cell apoptosis in SHAM group, IRI group and IRI/Cur group. Red arrows indicate TUNEL-positive cells. (B) Apoptosis indexes of above groups. n = 8 per group. IRI, ischemia/reperfusion injury; Cur, curcumin. *p < 0.05 vs. SHAM group; #p < 0.05 vs. IRI group.

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  Fig. 4 Protein levels of caspae-3 and caspase-9 in IRI rats with or without curcumin treatment. (A) The protein levels of caspase-3/-9 and cleaved caspase-3/-9 in SHAM group, IRI group, and IRI/Cur group. (B) Quantification results of Western blotting. n = 8 per group. IRI, ischemia/reperfusion injury; Cur, curcumin. *p < 0.05 vs. SHAM group; #p < 0.05 vs. IRI group.

• 

  Fig. 5 Histone acetylation near the promoter region of caspae-3 and caspase-9. ChiP assay was used to detect acH3K9 level in the upstream region of (A) caspase-3 (1,000 bp) and (B) caspase-9 (1,000 bp). The enrichment of CREB-binding protein (CBP)/p300 in the promoter region of (C) caspase-3 and (D) caspase-9 in SHAM group, IRI group and IRI/Cur group. (E) The protein levels of p300 and CBP in above groups. (F) Quantification results of western blotting. Results are expressed as mean ± SD. Statistical significance was determined by One-way ANOVA followed by Fisher’s Least Significant Difference test. n = 8 per group. IRI, ischemia/reperfusion injury; Cur, curcumin. *p < 0.05 vs. SHAM group; #p < 0.05 vs. IRI group.

• 

  Fig. 6 c-Jun N-terminal kinase (JNK) and curcumin show similar effects on p300/CREB-binding protein (CBP) and caspase3/9 expression. (A) The protein levels of caspase-3/-9 and cleaved caspase-3/-9 in SHAM group, IRI group, IRI/SP group and IRI/Cur group. (B) The level of histone acetylation related proteins including p300/CBP and acetyl-histone H3 in above groups. (C) The protein level of phosphorylated JNK in above groups. Results are expressed as mean ± SD. Statistical significance was determined by One-way ANOVA followed by Fisher’s Least Significant Difference test. n = 8 per group. IRI, ischemia/reperfusion injury; SP, SP600125; Cur, curcumin. *p < 0.05 vs. SHAM group; #p < 0.05 vs. IRI group.

• 

  Fig. 7 c-Jun N-terminal kinase (JNK) and curcumin show similar effects on H3K9 acetylation near the promoter region of caspase-3/-9. The enrichment of acH3K9 in the promoter regions of (A) caspase-3 and (B) caspase-9 in SHAM group, IRI group, IRI/SP group and IRI/Cur group. The binding level of CREB-binding protein (CBP)/p300 to the promoter region of (C) caspase-3 and (D) caspase-9 in above groups. (E) histone acetyltransferase (HAT) activity in above groups. Results are expressed as mean ± SD. Statistical significance was determined by One-way ANOVA followed by Fisher’s Least Significant Difference test. n = 8 per group. IRI, ischemia/reperfusion injury; SP, SP600125; Cur, curcumin. *p < 0.05 vs. SHAM group; #p < 0.05 vs. IRI group.

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  Fig. 8 Schematic illustration of the protective mechanism of curcumin against renal ischemia/reperfusion injury (IRI). The mechanism involves suppression on activation of c-Jun N-terminal kinase (JNK) pathway via epigenetic regulation of p300/CREB-binding protein (CBP)-mediated histone acetylation. In the IRI rat model, JNK activation promotes the elevation in acH3K9 level at caspase-3/-9 gene promoters, as well as the occupancy of p300/CBP on the gene promoters. These events can enhance caspase-3/-9 expression, thereby resulting in renal cell apoptosis involved in IRI. Curcumin can attenuate these changes through its effects on JNK signaling and histone acetylation.

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