Abstract
Inflammation is characterized by an increase in vascular permeability leading to edema formation. Phosphoinositide 3-kinase (PI3K) signaling pathway plays the main role to regulate several key events in the inflammatory response to damage and infection. Our previous in silico study showed that anthocyanin and ternatin flavonoids may offer antiinflammatory potential via molecular docking model. Here, we explored whether Clitoria ternatea anthocyanin extract (CTA) had protective activity against inflammation via regulating gene expression related to PI3Ks in rat edema. CTA was extracted in methanol and acetone using maceration and lyophilization. Edema model was done by injecting carrageenan into rat paw. Oral supplementation of CTA (100, 250, and 500 mg/kg BW) and ibuprofen standard (15 mg/kg BW) was given to rats daily for 7 days. After sample treatment, the right hind paw of the rat was injected with carrageenan to induce edema condition. The ability of CTA to suppress the inflammation in paw edema was measured by quantifying the increased percentage of rat paw volume at 1, 3, and 5 hrs after injection. Rat paw tissue was collected, and gene expression related to PI3Ks, i.e. Akt1, PKB, Ilk, Pdk2, Pik3Ca, and p53 was determined using quantitative PCR (qPCR). Carrageenan showed a time-dependent increase in volume of rat paw starting from injection until the next 5 hrs. It reached maximum volume increase of rat paw at 3 hrs after injection. CTA treatment had a dose-dependent pattern in reducing the edema volume in carrageenan-induced paw edema. At 500 mg/kg BW, CTA significantly suppressed the inflammation in paw edema compared to that of positive control, indicating by the low percentage of paw edema volume at 1, 3, and 5 hrs after carrageenan injection. At gene level, qPCR data showed that CTA significantly down-regulated the expression of genes involved in PI3Ks, such as Akt1, PKB, Ilk, Pdk2, Pik3Ca, and p53 in paw tissues. Thus, CTA may have a protective effect against edema via suppressing genes of PI3Ks.
Highlights
Inflammation is a body defense reaction to eliminate or limit the spread of injurious agent
Several pro-inflammatory mediators were released during the inflammatory response, including COX-2, TNF-α, NFκB, IL-7, and IL-6 and the induction of these proinflammatory mediators are largely regulated by transcriptional activation (Rommel and Camps, 2007)
We evaluated whether Clitoria ternatea anthocyanin extract (CTA) exerts its antiinflammatory effect in association with Phosphoinositide 3-kinase (PI3K) signaling pathway by evaluating genes that are involved in the pathway, such as Akt1, PKB, p53, Ilk, Pdk2, and Pik3Ca in carrageenan-induced paw edema in rats
Summary
Inflammation is a body defense reaction to eliminate or limit the spread of injurious agent. Acute inflammatory response is characterized by an increase in vascular permeability and cellular infiltration leading to edema formation, as a result of extravasation of fluid and proteins and accumulation of leukocytes at the inflammatory site. It is a defense mechanism, the complex events and mediators involved in the inflammatory reaction can induce, maintain or aggravate several diseases (Amdekar et al, 2012). Inhibitors of PI3K disrupt the accumulation of PIP3which in turn down-regulate the pro-inflammatory cytokines, thereby limit the inflammation, increase macrophage phagocytic ability, and resistance to infection (Koyasu, 2003). C. ternatea anthocyanin extract (CTA) was evaluated for anti-inflammatory activities to reduce paw edema volume by suppressing gene expression related to PI3K signaling in carrageenan-induced rat paw edema
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