肾损伤因子-1在脂多糖诱导的HK-2细胞炎症反应中的作用

doi:10.3969/j.issn.2095-9400.2019.02.002

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Abstract Objective: To investigate the expression of kidney injury molecule-1 (KIM-1) in lipopolysaccharide (LPS)-induced HK-2 cells and explore the biological process it maybe involved. Methods: Human renal tubular epithelial HK-2 cells were treated with LPS to establish the sepsis-induced renal cell inflammatory model. CCK-8 assay was used to detect the growth inhibition effect of LPS on HK-2 cells. The mRNA and protein expression levels of KIM-1 were detected by RT-PCR and Western blot respectively. The siRNA sequences targeted on KIM-1 gene were designed and tranfected to HK-2 cells, and then the effects of LPS on the growth inhibition of transfected HK-2 cells were detected by CCK-8 assays. Hoechst33342/PI staining was performed to detect the apoptosis in siRNA and control groups. Results: The proliferation of HK-2 cells was inhibited after treatment with LPS at final concentration of 50 μg/mL and 100 μg/mL for 24 h and 48 h, and the differences were statistically significant compared with the control group (P<0.05). The mRNA and protein levels of KIM-1 and IL-6 gene were greatly up-regulated after treatment with LPS in a dose-dependent manner, the differences being statistically significant (P<0.05). The proportion of Hoechst3342 positive cells in LPS treatment group was significantly higher than that in control group. The expression levels of KIM-1 and IL-6 genes in siRNA groups were significantly lower than that in siRNA-NC group, with significant differences (P<0.05); The proliferation rate was significantly higher in siRNA transfection group, compared with the siRNA-NC group, and the differences were statistically significant (P<0.05). The fluorescence intensity of Hoechst3342 staining in siRNA transfection groups was lower than that in siRNA-NC group, suggesting that siRNA transfection targeting on KIM-1 gene could inhibit the apoptosis induced by LPS. Conclusion: LPS can induce the proliferation inhibition and apoptosis of HK-2 cells, and up-regulate the expression of KIM-1 and IL-6 genes. KIM-1 may participate in the inflammatory reaction by regulating the process of apoptosis and cell growth inhibition.

Key words： lipopolysaccharide kidney injury molecule-1 HK-2 cells cellular inflammatory reaction

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	JIANG Yingying
	CHEN Longwang
	LIN Yue
	CHEN Xinguo
	ZHEGN Yanyan
	LU Zhongqiu

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https://xb.wmu.edu.cn/EN/10.3969/j.issn.2095-9400.2019.02.002 OR https://xb.wmu.edu.cn/EN/Y2019/V49/I2/85

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