外源性趋化因子在高糖环境下对人脐静脉内皮细胞功能的影响
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上海市重点实验室资助项目(No.17DZ2260100)


Effect of exogenous chemokines on the function of human umbilical vein endothelial cells under the environment of high glucose
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Science and Technology Commission of Shanghai(No.17DZ2260100)

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    摘要:

    目的:探讨高糖环境下外源性趋化因子CXCL9、CXCL10、CXCL11对人脐静脉内皮细胞(HUVEC)功能的影响及其机制。

    方法:将对数生长期的细胞分为对照组(葡萄糖浓度5.5mmol/L)、低糖组(葡萄糖浓度5mmol/L)、高糖组(葡萄糖浓度30mmol/L),分别加入CXCL9(100ng/mL)、CXCL10(10ng/mL)、CXCL11(100ng/mL),培养24、48、72h,采用CCK-8法检测细胞增殖能力,RT-PCR检测CXCR3 mRNA的表达,免疫荧光法检测细胞增殖标志分子Ki-67阳性表达情况。

    结果:CCK-8法检测结果显示,加入三种外源性趋化因子后,随着时间的延长,对照组细胞吸光度值逐渐增强; 低糖组细胞吸光度值呈现先增强后降低的趋势,48h达到高峰; 高糖组细胞吸光度值总体呈降低趋势。RT-PCR检测结果显示,加入三种外源性趋化因子后48、72h,低糖组和高糖组细胞CXCR3 mRNA表达水平均高于对照组,且均较同组24h时升高。免疫荧光检测结果显示,加入三种外源性趋化因子后72h,低糖组与高糖组细胞Ki-67荧光强度降低,高糖组变化更明显。

    结论:高糖环境下外源性加入CXCL9、CXCL10、CXCL11可使HUVEC细胞活力下降并诱导CXCR3表达增强,且以外源性加入CXCL10、CXCL11配体后,CXCR3表达增幅最高,这可能成为临床干预糖尿病视网膜病变的靶点。

    Abstract:

    AIM: To investigate the effects of exogenous CXCL9, CXCL10 and CXCL11 on human umbilical vein endothelial cells under a high glucose environment and explore its mechanisms.

    METHODS: The cells in logarithmic growth stage were divided into control group(glucose concentration 5.5mmol/L), low glucose group(glucose concentration 5mmol/L), high glucose group(glucose concentration 30mmol/L). CXCL9(100ng/mL), CXCL10(10ng/mL)and CXCL11(100ng/mL)were added respectively, cultured for 24, 48 and 72h. CCK-8 method was used to detect cell proliferation, RT-PCR was used to detect CXCR3 mRNA expression, and immunofluorescence was used to detect Ki-67 expression.

    RESULTS: The results of CCK-8 method showed that the absorbance value of the control group increased gradually with the increase of time after adding three exogenous chemokines. The absorbance value of the low glucose group increased first and then decreased, reaching the peak at 48h. The absorbance value of the high glucose group decreased generally. The results of RT-PCR showed that the expression of CXCR3 mRNA in low glucose group and high glucose group was higher than that in 24h after adding CXCL9, CXCL10 and CXCL11 for 48 and 72h. The results of immunofluorescence showed that the fluorescence intensity of Ki-67 decreased in the low and high glucose 72h after adding CXCL9, CXCL10 and CXCL11. The change in the high glucose group is more obvious.

    CONCLUSION: Exogenous CXCL9, CXCL10 and CXCL11 can decrease the activity of human umbilical vein cell under high glucose environments and induce the increase in CXCR3 expression. The increase of CXCR3 reached the highest after adding exogenous CXCL10 and CXCL11, suggesting a target for clinical intervention of diabetic retinopathy.

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胡海林,罗敏.外源性趋化因子在高糖环境下对人脐静脉内皮细胞功能的影响.国际眼科杂志, 2020,20(5):773-777.

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  • 收稿日期:2019-06-04
  • 最后修改日期:2020-04-15
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  • 在线发布日期: 2020-04-26
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