Abstract:
Objective To study the effect and mechanism of monocyte chemotactic protein-1 (MCP-1)/transforming growth factor beta1(TGF-β1)/collagen-Ⅰ (COL-Ⅰ) pathways for human kidney 2 (HK-2) cells induced by high glucose.
Methods Methyl Thiazolyl Tetrazolium (MTT) method was used to establish HK-2 cell model damaged by sugar. The protein expression levels of MCP-1, TGF-β1 and COL-Ⅰ in supernatant of HK-2 cells were determined by enzyme-linked immunosorbent assay (ELISA), and the expressions of MCP-1 , TGF-β1 and COL-ⅠmRNA in HK-2 cells were determined by reverse transcription-polymerase chain reaction (RT-PCR) after treatment with different concentrations of alpha linolenic acid (ALA)/linoleic acid (LA).
Results After 48 h intervention in HK-2 cells induced by glucose were treated with 50 μmol/L ALA and LA and 50 μmol/L mixture of ALA/LA with the ratio of 1 to 4, the expression levels of MCP-1mRNA and MCP-1 protein were significantly decreased (P < 0.05). After HK-2 cells induced by glucose and treated with 100 μmol/L ALA and LA for 48 h, TGF-β1mRNA and TGF-β1 protein expression were significantly decreased(P < 0.05). After HK-2 cells induced by glucose and treated with 100 μmol/L ALA, 50 μmol/L LA and 50 μmol/L mixture of ALA/LA with the ratio of 1 ∶ 4 for 48 h, the expression of COL-Ⅰ mRNA and COL-Ⅰ protein were significantly decreased (P < 0.05).
Conclusion HK-2 cells are damaged by high glucose, manifesting as increased expression of cytokines MCP-1 and TGF-β1, and increased expression of fibrosis product COL-Ⅰ. ALA/LA can down-regulate the expression of MCP-1 and TGF-β1, reduce the level of COL-Ⅰ, and protect HK-2 cells damaged by glucose.
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