Cinnamaldehyde protects against rat intestinal ischemia/reperfusion injuries by synergistic inhibition of NF-κB and p53

Abstract

Our preliminary study shows that cinnamaldehyde (CA) could protect against intestinal ischemia/reperfusion (I/R) injuries, in which p53 and NF-κB p65 play a synergistic role. In this study, we conducted in vivo and in vitro experiments to verify this proposal. SD rats were pretreated with CA (10 or 40 mg · kg −1 · d −1 , ig) for 3 days, then subjected to 1 h mesenteric ischemia followed by 2 h reperfusion. CA pretreatment dose-dependently ameliorated morphological damage and reduced inflammation evidenced by decreased TNF-α, IL-1β, and IL-6 levels and MPO activity in I/R-treated intestinal tissues. CA pretreatment also attenuated oxidative stress through restoring SOD, GSH, LDH, and MDA levels in I/R-treated intestinal tissues. Furthermore, CA pretreatment significantly reduced the expression of inflammation/apoptosis-related NF-κB p65, IKKβ, IK-α, and NF-κB p50, and downregulated apoptotic protein expression including p53, Bax, caspase-9 and caspase-3, and restoring Bcl-2, in I/R-treated intestinal tissues. We pretreated IEC-6 cells in vitro with CA for 24 h, followed by 4 h hypoxia and 3 h reoxygenation (H/R) incubation. Pretreatment with CA (3.125, 6.25, and 12.5 μmol · L −1 ) significantly reversed H/R-induced reduction of IEC-6 cell viability. CA pretreatment significantly suppressed oxidative stress, NF-κB activation and apoptosis in H/R-treated IEC-6 cells. Moreover, CA pretreatment significantly reversed mitochondrial dysfunction in H/R-treated IEC-6 cells. CA pretreatment inhibited the nuclear translocation of p53 and NF-κB p65 in H/R-treated IEC-6 cells. Double knockdown or overexpression of p53 and NF-κB p65 caused a synergistic reduction or elevation of p53 compared with knockdown or overexpression of p53 or NF-κB p65 alone. In H/R-treated IEC-6 cells with double knockdown or overexpression of NF-κB p65 and p53, CA pretreatment caused neither further decrease nor increase of NF-κB p65 or p53 expression, suggesting that CA-induced synergistic inhibition on both NF-κB and p53 played a key role in ameliorating intestinal I/R injuries. Finally, we used immunoprecipitation assay to demonstrate an interaction between p53 and NF-κB p65, showing the basis for CA-induced synergistic inhibition. Our results provide valuable information for further studies.

Sesamin Protects against and Ameliorates Rat Intestinal Ischemia/Reperfusion Injury with Involvement of Activating Nrf2/HO‐1/NQO1 Signaling Pathway

“…DAPI staining is used to detect apoptosis in vitro [ 22 , 28 ]. The in situ observation in the IEC-6 battery configuration is shown in Figure 5(d) .…”

“…Histological evidence of intestinal I/R injury includes decreased villus height, increased cellular infiltrate, and mucosal sloughing [ 45 ]. And the increase of serum aminotransferase levels, such as AST and ALT, caused by remote organ injuries is also one of the characteristics of intestinal I/R injury [ 28 , 46 , 47 ]. Proinflammatory cytokines, such as TNF- α , IL-6, and IL-1 β , in serum of intestinal I/R animals are significantly increased [ 22 , 28 , 48 ], which is related to neutrophil activation and tissue damage caused by I/R [ 47 , 49 ].…”

Protective Effects of Cinnamaldehyde against Mesenteric Ischemia-Reperfusion-Induced Lung and Liver Injuries in Rats

“…Our previous study showed that mesenteric I/R induced excessive intestinal (local) morphological changes, inflammation, oxidative stress, apoptosis, and upregulated p53 and NF- κ B pathway-related proteins in mesenteric I/R-injured rats and hypoxia/reoxygenation- (H/R-) injured intestinal epithelial cells-6 (IEC-6), and CA pretreatment significantly restored all the above-mentioned changes in mesenteric I/R-treated rats and H/R-treated IEC-6 cells [ 18 ].…”

“…The rats were randomly assigned to 5 groups (5 rats/group): (1) sham group: via the intragastric gavage (ig) route, rats were given a vehicle daily for 3 days before rat sham surgery; (2) sham+CA group: rats were subjected to sham surgery after pretreatment with CA (ig) with the concentration 40 mg/kg daily/3 days; (3) I/R group: via the intragastric gavage (ig) route, rats were given a vehicle daily for 3 days before they were induced with 1 h mesenteric ischemia and then reperfusion for additional 2 h; (4) I/R+CA (L) group: rats were subjected to I/R surgery after they were pretreated with CA (ig) with the concentration 10 mg/kg/day/3 days [ 18 , 34 ]; and (5) I/R+CA (H) group: rats were subjected to I/R surgery after they were pretreated with CA (ig) with the concentration 40 mg/kg/day/3 days [ 18 , 35 , 36 ]. The intragastric gavage suspension of CA in carboxymethyl cellulose (1% CMC) was prepared daily and freshly and given at 2 mL/kg.…”

“…Irisin protects against mesenteric I/R-induced liver injury [ 17 ]. And in our previous study, CA attenuates mesenteric I/R-induced gut injury via a synergistic inhibition of p53/NF- κ B signaling pathways [ 18 ].…”