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ENGLISH ABSTRACT
JOURNAL ARTICLE
RESEARCH SUPPORT, NON-U.S. GOV'T
[Effects of leflunomide on podocytes exposed to high glucose condition and its mechanism].
Zhonghua Yi Xue za Zhi [Chinese medical journal] 2013 June 26
OBJECTIVE: To explore the protective effects of leflunomide (A771726) on the expression of podocalyxin, NF-κB and matrix metalloproteinase-9 (MMP-9) in podocytes exposed to high glucose environment and elucidate its mechanism.
METHODS: Podocytes were cultured in high glucose. And the altered expressions of podocyte protein podocalyxin were detected by Western blotting at different timepoints. Then podocytes were divided into 4 groups of normal glucose control, leflunomide, high glucose and hypertonic control. The expression level of podocalyxin protein in each group was detected by Western blotting. And NF-κB p65 and phosphorylation of NF-κB p65 (P-NF-κBp65) in podocytes cultured in high glucose were detected at different timepoints. And then the podocytes were divided into 5 groups of normal glucose, mannitol, hypertonic control, high glucose, leflunomide and PDTC (NF-κB blocker). And the expressions of MMP-9 protein in these groups were also detected by Western blotting.
RESULTS: In the high glucose environment, the expression of podocalyxin declined instantly. Compared with the high-glucose group, the podocalyxin expression of the leflunomide group was significantly higher than the high glucose group (0.46 ± 0.04 vs 0.13 ± 0.03, P < 0.05). After 30-minute stimulation by high glucose, the activation of NF-κB started and the expression of P-NF-κBp65 protein increased. Such activities peaked at 60 minutes and reverted to a basic level after 6 hours. Compared with the high glucose group, the expressions of MMP-9 in PDTC and leflunomide groups were significantly lower than the high glucose group. And the differences were statistically significant (0.71 ± 0.01, 0.64 ± 0.03 vs 1.64 ± 0.03, both P < 0.05).
CONCLUSIONS: Leflunomide has protective effects on podocytes in high glucose. And its mechanism is possibly due to a lowered expression of MMP-9 through an inhibition of NF-κB activation.
METHODS: Podocytes were cultured in high glucose. And the altered expressions of podocyte protein podocalyxin were detected by Western blotting at different timepoints. Then podocytes were divided into 4 groups of normal glucose control, leflunomide, high glucose and hypertonic control. The expression level of podocalyxin protein in each group was detected by Western blotting. And NF-κB p65 and phosphorylation of NF-κB p65 (P-NF-κBp65) in podocytes cultured in high glucose were detected at different timepoints. And then the podocytes were divided into 5 groups of normal glucose, mannitol, hypertonic control, high glucose, leflunomide and PDTC (NF-κB blocker). And the expressions of MMP-9 protein in these groups were also detected by Western blotting.
RESULTS: In the high glucose environment, the expression of podocalyxin declined instantly. Compared with the high-glucose group, the podocalyxin expression of the leflunomide group was significantly higher than the high glucose group (0.46 ± 0.04 vs 0.13 ± 0.03, P < 0.05). After 30-minute stimulation by high glucose, the activation of NF-κB started and the expression of P-NF-κBp65 protein increased. Such activities peaked at 60 minutes and reverted to a basic level after 6 hours. Compared with the high glucose group, the expressions of MMP-9 in PDTC and leflunomide groups were significantly lower than the high glucose group. And the differences were statistically significant (0.71 ± 0.01, 0.64 ± 0.03 vs 1.64 ± 0.03, both P < 0.05).
CONCLUSIONS: Leflunomide has protective effects on podocytes in high glucose. And its mechanism is possibly due to a lowered expression of MMP-9 through an inhibition of NF-κB activation.
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