白细胞介素10对HK-2细胞转分化的影响及其机制的初步探讨
发布时间：2018-07-27 13:14
【摘要】： 目的探讨白细胞介素10对肾小管上皮细胞转分化的影响,并初步讨论其作用机制。 方法本实验采用HK-2细胞为靶细胞。将HK-2细胞完全随机分为5组:①对照组:5.5mmol/L的D-葡萄糖组;②高糖组:30mmol/L的D-葡萄糖组;③l ng/ml IL-l0组:30mmol/L的D-葡萄糖+ l ng/ml IL-l0组;④5 ng/ml IL-l0组:30mmol/L的D-葡萄糖+ 5 ng/ml IL-l0组;⑤25 ng/ml IL-l0组:30mmol/L的D-葡萄糖+ 25 ng/ml IL-l0组。培养48h后,倒置显微镜观察HK-2细胞的形态变化;免疫细胞化学法检测各组细胞E-cad、α-SMA蛋白的表达;RT-PCR法检测各组细胞PDGF mRNA、CTGF mRNA的表达;western blot法检测各组细胞PDGF、E-cad、α-SMA蛋白的表达;ELISA法检测各组细胞上清液中FN的含量。 结果(1)细胞形态:对照组细胞呈鹅卵石样紧密排列生长;高糖组部分细胞间隙增大,和附近的细胞脱离接触,失去鹅卵石样形态,呈梭形;l ng/ml组、5 ng/ml组、25 ng/ml IL-l0组细胞形态介于对照组和高糖组之间。(2)高糖组与对照组相比,E-cad表达减少,PDGF、CTGF、α-SMA和FN表达增加,其差异均有显著性(P0.05),提示高糖可诱导HK-2细胞转分化。(3)l ng/ml组、5 ng/ml组、25 ng/ml IL-l0组与高糖组相比,E-cad表达增加,PDGF、CTGF、α-SMA和FN表达减少,其差异均有显著性(P0.05),提示IL-10可以抑制HK-2细胞转分化。 结论高糖可通过诱导PDGF和CTGF的表达使HK-2细胞E-cad表达减少、α-SMA和FN表达增加,从而诱导HK-2细胞转分化及细胞外基质产生;IL-10可以通过抑制PDGF和CTGF的表达从而抑制高糖诱导的HK-2细胞转分化及细胞外基质产生。
[Abstract]:Objective to investigate the effect of interleukin 10 (IL 10) on renal tubular epithelial cell transdifferentiation and its mechanism. Methods HK-2 cells were used as target cells. The HK-2 cells were randomly divided into 5 groups: 1 control group: 5.5 mmol / L D- glucose group; (2) high glucose group: 30 mmol / L D- glucose group; 30 mmol / L D- glucose group; 1 / 30 mmol / L D- glucose / L ng/ml IL-l0 group; 45 ng/ml IL-l0 group: 30 mmol / L D- glucose 5 ng/ml IL-l0 group; Ng/ml IL-l0 group: 30 mmol / L D- glucose 25 ng/ml IL-l0 group. After 48 hours of culture, the morphological changes of HK-2 cells were observed by inverted microscope. The expression of E-cadand 伪 -SMA protein was detected by immunocytochemistry and the expression of PDGF mRNA-CTGF mRNA was detected by RT-PCR. The expression of 伪 -SMA protein in the supernatant of each group was detected by Western blot, and the expression of 伪 -SMA protein was detected by Elisa. Results (1) Cell morphology: in the control group, the cells were closely arranged in pebbles, while in the high glucose group, the gap between the cells and the nearby cells was increased, and the pebble-like morphology was lost. The expression of E-cad, 伪 -SMA and FN in the high glucose group was significantly lower than that in the control group, and the expression of E-cad, 伪 -SMA and FN in the high glucose group was significantly lower than that in the control group, and the expression of 伪 -SMA and FN was increased in the high glucose group compared with the control group. The differences were significant (P0.05), suggesting that high glucose could induce the transdifferentiation of HK-2 cells. (3) the expression of E-cad increased in the 25 ng/ml IL-l0 group of 5 ng/ml group compared with the high glucose group, and the expression of E-cad, 伪 -SMA and FN decreased significantly (P0.05), suggesting that IL-10 could inhibit the transdifferentiation of HK-2 cells. Conclusion High glucose can reduce the expression of E-cad and increase the expression of 伪 -SMA and FN in HK-2 cells by inducing the expression of PDGF and CTGF. Thus, the transdifferentiation of HK-2 cells and the production of IL-10 in extracellular matrix can inhibit the transdifferentiation and extracellular matrix production of HK-2 cells induced by high glucose by inhibiting the expression of PDGF and CTGF.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R392.11
本文编号：2147951
[Abstract]:Objective to investigate the effect of interleukin 10 (IL 10) on renal tubular epithelial cell transdifferentiation and its mechanism. Methods HK-2 cells were used as target cells. The HK-2 cells were randomly divided into 5 groups: 1 control group: 5.5 mmol / L D- glucose group; (2) high glucose group: 30 mmol / L D- glucose group; 30 mmol / L D- glucose group; 1 / 30 mmol / L D- glucose / L ng/ml IL-l0 group; 45 ng/ml IL-l0 group: 30 mmol / L D- glucose 5 ng/ml IL-l0 group; Ng/ml IL-l0 group: 30 mmol / L D- glucose 25 ng/ml IL-l0 group. After 48 hours of culture, the morphological changes of HK-2 cells were observed by inverted microscope. The expression of E-cadand 伪 -SMA protein was detected by immunocytochemistry and the expression of PDGF mRNA-CTGF mRNA was detected by RT-PCR. The expression of 伪 -SMA protein in the supernatant of each group was detected by Western blot, and the expression of 伪 -SMA protein was detected by Elisa. Results (1) Cell morphology: in the control group, the cells were closely arranged in pebbles, while in the high glucose group, the gap between the cells and the nearby cells was increased, and the pebble-like morphology was lost. The expression of E-cad, 伪 -SMA and FN in the high glucose group was significantly lower than that in the control group, and the expression of E-cad, 伪 -SMA and FN in the high glucose group was significantly lower than that in the control group, and the expression of 伪 -SMA and FN was increased in the high glucose group compared with the control group. The differences were significant (P0.05), suggesting that high glucose could induce the transdifferentiation of HK-2 cells. (3) the expression of E-cad increased in the 25 ng/ml IL-l0 group of 5 ng/ml group compared with the high glucose group, and the expression of E-cad, 伪 -SMA and FN decreased significantly (P0.05), suggesting that IL-10 could inhibit the transdifferentiation of HK-2 cells. Conclusion High glucose can reduce the expression of E-cad and increase the expression of 伪 -SMA and FN in HK-2 cells by inducing the expression of PDGF and CTGF. Thus, the transdifferentiation of HK-2 cells and the production of IL-10 in extracellular matrix can inhibit the transdifferentiation and extracellular matrix production of HK-2 cells induced by high glucose by inhibiting the expression of PDGF and CTGF.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R392.11
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2 李涛,冷希圣,秦致中,宋盛晗,赵力,熊亮发,彭吉润;白细胞介素10对肝星状细胞激活的调节[J];中华肝脏病杂志;2005年01期
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