Limits...
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.

Lee J, Yang EJ, Shin JS, Kim DH, Lee SS, Choi IH - Immune Netw (2011)

Bottom Line: Since CKD-712 has been developed as an anti-inflammatory agent, we examined the effect of CKD-712 during TLR4 signaling.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.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, Brain Korea 21 Project for Medical Science, Institute for Immunology and Immunological Diseases, Yonsei University College of Medicine, Seoul 120-752, Korea.

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.

No MeSH data available.


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.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3275714&req=5

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.

Mentions: Next, we investigated whether treatment with CKD-712 modulates MAP kinaseactivation. After stimulation with LPS, CKD-712 did not inhibit activation of ERK, JNK or p38 kinase in HEK293-TLR4 cells (Fig. 2). Rather, pre-treatment with CKD-712 delayed activation of these kinases. As shown in Fig. 3, MAP kinases were phosphorylated 15~30 min following stimulation with LPS, after which activation subsided in 60 min. However, CKD-712 treatment extended the activation of ERK and JNK, which were found to be activated 60 min after TLR4 stimulation (Fig. 2). Phosphorylation of IRF3 was not inhibited and showed a sustained activation like the MAP kinases (Fig. 2). Interestingly, the activation of Akt was augmented slightly with pre-treatment of CKD-712 (50µM) after LPS stimulation (Fig. 3, left). But higher concentration of CKD-712 (100µM) inhibited the activation of Akt completely (Fig. 3, right).


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.

Lee J, Yang EJ, Shin JS, Kim DH, Lee SS, Choi IH - Immune Netw (2011)

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.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3275714&req=5

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.
Mentions: Next, we investigated whether treatment with CKD-712 modulates MAP kinaseactivation. After stimulation with LPS, CKD-712 did not inhibit activation of ERK, JNK or p38 kinase in HEK293-TLR4 cells (Fig. 2). Rather, pre-treatment with CKD-712 delayed activation of these kinases. As shown in Fig. 3, MAP kinases were phosphorylated 15~30 min following stimulation with LPS, after which activation subsided in 60 min. However, CKD-712 treatment extended the activation of ERK and JNK, which were found to be activated 60 min after TLR4 stimulation (Fig. 2). Phosphorylation of IRF3 was not inhibited and showed a sustained activation like the MAP kinases (Fig. 2). Interestingly, the activation of Akt was augmented slightly with pre-treatment of CKD-712 (50µM) after LPS stimulation (Fig. 3, left). But higher concentration of CKD-712 (100µM) inhibited the activation of Akt completely (Fig. 3, right).

Bottom Line: Since CKD-712 has been developed as an anti-inflammatory agent, we examined the effect of CKD-712 during TLR4 signaling.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.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, Brain Korea 21 Project for Medical Science, Institute for Immunology and Immunological Diseases, Yonsei University College of Medicine, Seoul 120-752, Korea.

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.

No MeSH data available.