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. 2011 Dec;11(6):420-3.
doi: 10.4110/in.2011.11.6.420. Epub 2011 Dec 31.

CKD-712, (S)-1-(α-naphthylmethyl)-6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline, Inhibits the NF-κB Activation and Augments Akt Activation during TLR4 Signaling

Jeonggi Lee  1 , Eun-Jeong Yang, Jeon-Soo Shin, Dal-Hyun Kim, Sung-Sook Lee, In-Hong Choi
Affiliations

CKD-712, (S)-1-(α-naphthylmethyl)-6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline, Inhibits the NF-κB Activation and Augments Akt Activation during TLR4 Signaling

Jeonggi Lee et al. Immune Netw. 2011 Dec.

Abstract

Since CKD-712 has been developed as an anti-inflammatory agent, we examined the effect of CKD-712 during TLR4 signaling. Using HEK293 cells expressing TLR4, CKD-712 was pre-treated 1 hr before LPS stimulation. Activation of NF-κB was assessed by promoter assay. The activation of ERK, JNK, p38, IRF3 and Akt was measured by western blotting. CKD-712 inhibited the NF-κB signaling triggered by LPS. The activation of ERK, JNK, p38 or IRF3 was not inhibited by CKD-712. On the contrary the activation of these molecules was augmented slightly. The activation of Akt with stimulation of LPS was also enhanced with CKD-712 pre-treatment at lower concentration, but was inhibited at higher concentration. We suggest that during TLR4 signaling CKD-712 inhibits NF-κB activation. However, CKD-712 augmented the activation of Akt as well as Map kinases. Therefore, we suggest that CKD-712 might have a role as an immunomodulator.

Keywords: Akt; CKD-712; TLR4; immunomodulator.

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Conflict of interest statement

The authors have no financial conflict of interest.

Figures

Figure 1
Figure 1
Effect of CKD-712 on TLR4 signaling. (A) To assess CKD-712 toxicity HEK293-TLR4 cells (2×105) were cultured with CKD-712 for 24 hrs and cell viability was determined by MTT assay. (B) To evaluate the effect of CKD-712 on NF-κB activation 2×105 HEK-TLR4 cells/well were seeded in 12-well plates and were cultured for 2 days. Then NF-κB promoter and pCMV β-gal vector were co-transfeceted overnight and CKD-712 (50µM) was pre-treated before LPS treatment. LPS was treated for 6 hrs. After measuring luciferase activity and β-galactosidase activity the luciferase activity was normalized for transfection efficiency with the β-galactosidase activity.
Figure 2
Figure 2
Effect of CKD-712 on activation of MAP kinases and IRF3 in TLR4 signaling. HEK293-TLR4 cells were stimulated with LPS (50 ng/ml). Western blot was performed anti-ERK, anti-JNK, anti-p38 or anti-IRF3 antibodies. CKD-712 (50µM) was pre-treated 1 hr before LPS stimulation. Three independent experiments were performed and the representative data are shown.
Figure 3
Figure 3
Effect of CKD-712 on Akt activation in TLR4 signaling. HEK293-TLR4 cells were stimulated with LPS (50 ng/mL) and western blot was performed with anti-Akt antibodies. CKD-712 (50 or 100µM) was pretreated 1 hr before LPS stimulation. Two independent experiments were performed and the representative data are shown. The density was measures by densitometry and relative ratio of p-Akt/Akt in study groups was calculated based on the p-Akt/Akt of control group as 1.0.
See this image and copyright information in PMC

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