P38MAPK抑制剂SB203580对高糖诱导HK-2细胞转分化的影响
目的探讨不同浓度P38丝裂原活化蛋白激酶（P38MAPK）抑制剂SB203580在高糖诱导肾小管上皮细胞-肌成纤维细胞转分化（TEMT）过程中的机制及其较佳作用浓度。方法体外培养人近端肾小管上皮细胞（HK-2）并分为对照组（5.5 mmol/L GS）、DMSO组（5.5 mmol/L GS＋30μmol/L SB203580等体积的DMSO）、高糖组（30mmol/L GS）,以及30 mmol/L GS＋5、10、20、30μmol/LSB203580处理的S5、S10、S20及S30组,干预48 h。四甲基偶氮唑蓝（MTT）法检测细胞增殖情况,计算半数抑制浓度（IC50）;选取对照组、高...
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| Published in | 天津医药 Vol. 44; no. 4; pp. 426 - 429 |
|---|---|
| Main Author | |
| Format | Journal Article |
| Language | Chinese |
| Published |
石河子大学第一附属医院肾病科 邮编832008
2016
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| Subjects | |
| Online Access | Get full text |
| ISSN | 0253-9896 |
| DOI | 10.11958/20150004 |
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| Abstract | 目的探讨不同浓度P38丝裂原活化蛋白激酶（P38MAPK）抑制剂SB203580在高糖诱导肾小管上皮细胞-肌成纤维细胞转分化（TEMT）过程中的机制及其较佳作用浓度。方法体外培养人近端肾小管上皮细胞（HK-2）并分为对照组（5.5 mmol/L GS）、DMSO组（5.5 mmol/L GS＋30μmol/L SB203580等体积的DMSO）、高糖组（30mmol/L GS）,以及30 mmol/L GS＋5、10、20、30μmol/LSB203580处理的S5、S10、S20及S30组,干预48 h。四甲基偶氮唑蓝（MTT）法检测细胞增殖情况,计算半数抑制浓度（IC50）;选取对照组、高糖组、S30组,Western blot法检测P38MAPK、P-P38MAPK及α-平滑肌肌动蛋白（SMA）的表达、免疫荧光法检测α-SMA的表达。结果 （1）与对照组相比,DMSO对HK-2细胞增殖无显著抑制作用（P〉0.05）;高糖组、S5组HK-2细胞增殖增多（P〈0.05）;S20、S30组HK-2细胞增殖减少（P〈0.05）。与高糖组相比,S5、S10、S20、S30组细胞增殖均受到抑制（P〈0.05）。（2）与对照组相比,高糖组、S30组P-P38MAPK表达量增高（P〈0.05）。与高糖组相比,S30组P-P38MAPK的表达量降低（P〈0.05）。3组P38MAPK表达量无显著差异（P〉0.05）。（3）与对照组相比,高糖组、S30组α-SMA表达量增高（P〈0.05）。与高糖组相比,S30组α-SMA表达量降低（P〈0.05）。结论 30 mmol/L GS可以诱导HK-2细胞TEMT;30μmol/L SB203580是抑制HK-2细胞TEMT的较佳抑制浓度,SB203580可能通过下调P-P38MAPK表达,从而抑制HK-2细胞增殖及胞浆中α-SMA的表达,延缓TEMT进程。 |
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| AbstractList | R692; 目的：探讨不同浓度P38丝裂原活化蛋白激酶（P38MAPK）抑制剂SB203580在高糖诱导肾小管上皮细胞-肌成纤维细胞转分化（TEMT）过程中的机制及其较佳作用浓度。方法体外培养人近端肾小管上皮细胞（HK-2）并分为对照组（5.5 mmol/L GS）、DMSO组（5.5 mmol/L GS+30μmol/L SB203580等体积的DMSO）、高糖组（30 mmol/L GS），以及30 mmol/L GS+5、10、20、30μmol/LSB203580处理的S5、S10、S20及S30组，干预48 h。四甲基偶氮唑蓝（MTT）法检测细胞增殖情况，计算半数抑制浓度（IC50）；选取对照组、高糖组、S30组，Western blot法检测P38MAPK、P-P38MAPK及α-平滑肌肌动蛋白（SMA）的表达、免疫荧光法检测α-SMA的表达。结果（1）与对照组相比，DMSO对HK-2细胞增殖无显著抑制作用（P>0.05）；高糖组、S5组HK-2细胞增殖增多（P<0.05）；S20、S30组HK-2细胞增殖减少（P<0.05）。与高糖组相比，S5、S10、S20、S30组细胞增殖均受到抑制（P<0.05）。（2）与对照组相比，高糖组、S30组P-P38MAPK表达量增高（P<0.05）。与高糖组相比，S30组P-P38MAPK的表达量降低（P<0.05）。3组P38MAPK表达量无显著差异（P>0.05）。（3）与对照组相比，高糖组、S30组α-SMA表达量增高（P<0.05）。与高糖组相比，S30组α-SMA表达量降低（P<0.05）。结论30 mmol/L GS可以诱导HK-2细胞TEMT；30μmol/L SB203580是抑制HK-2细胞TEMT的较佳抑制浓度，SB203580可能通过下调P-P38MAPK表达，从而抑制HK-2细胞增殖及胞浆中α-SMA的表达，延缓TEMT进程。 目的探讨不同浓度P38丝裂原活化蛋白激酶（P38MAPK）抑制剂SB203580在高糖诱导肾小管上皮细胞-肌成纤维细胞转分化（TEMT）过程中的机制及其较佳作用浓度。方法体外培养人近端肾小管上皮细胞（HK-2）并分为对照组（5.5 mmol/L GS）、DMSO组（5.5 mmol/L GS＋30μmol/L SB203580等体积的DMSO）、高糖组（30mmol/L GS）,以及30 mmol/L GS＋5、10、20、30μmol/LSB203580处理的S5、S10、S20及S30组,干预48 h。四甲基偶氮唑蓝（MTT）法检测细胞增殖情况,计算半数抑制浓度（IC50）;选取对照组、高糖组、S30组,Western blot法检测P38MAPK、P-P38MAPK及α-平滑肌肌动蛋白（SMA）的表达、免疫荧光法检测α-SMA的表达。结果 （1）与对照组相比,DMSO对HK-2细胞增殖无显著抑制作用（P〉0.05）;高糖组、S5组HK-2细胞增殖增多（P〈0.05）;S20、S30组HK-2细胞增殖减少（P〈0.05）。与高糖组相比,S5、S10、S20、S30组细胞增殖均受到抑制（P〈0.05）。（2）与对照组相比,高糖组、S30组P-P38MAPK表达量增高（P〈0.05）。与高糖组相比,S30组P-P38MAPK的表达量降低（P〈0.05）。3组P38MAPK表达量无显著差异（P〉0.05）。（3）与对照组相比,高糖组、S30组α-SMA表达量增高（P〈0.05）。与高糖组相比,S30组α-SMA表达量降低（P〈0.05）。结论 30 mmol/L GS可以诱导HK-2细胞TEMT;30μmol/L SB203580是抑制HK-2细胞TEMT的较佳抑制浓度,SB203580可能通过下调P-P38MAPK表达,从而抑制HK-2细胞增殖及胞浆中α-SMA的表达,延缓TEMT进程。 |
| Abstract_FL | Objective To observe the effects of different concentrations of SB203580, the inhibitor of P38MAPK, in process of high glucose (GS)-induced renal tubular epithelial-myofibroblast transdifferentiation (TEMT). Methods The cultured human renal tubular epithelial cells (HK-2) were divided into control group (5.5 mmol/L GS), GS (30 mmol/L GS) group and different concentrations of SB203580 (30 mmol/L GS +5, 10, 20 and 30 μmol/L SB203580) groups. The treat?ments were for 48 hours. MTT assay was used to observe cell proliferation. The median inhibiting concentration (IC50) was cal?culated. Western blot assay was used to detect the expressions of P38MAPK, P-P38MAPK andα-smooth muscle actin (α-SMA) in control group, high-glucose group and S30 group. The expression ofα-SMA was also detected by the method of im?munofluorescence. Results 1.Compared with control group, there was no significant inhitory effect on proliferation rate in DMSO group (P>0.05). There were increased HK-2 cells in high glucose group and S5group (P<0.05). Proliferation rates were significantly decreased in S20 and S30 groups (P<0.05). Compared with high glucose group, the proliferation rates of HK-2 cells were inhibited in S5, S10, S20 and S30 groups (P<0.05). 2. The expression of P-P38MAPK was significantly higher in high glucose group and S30 group than that of control group (P<0.05). Compared with high glucose group, the ex?pression of P-P38MAPK was significantly decreased in S30 group (P<0.05), whereas no significant difference in the expres?sion of P38MAPK between the two groups (P>0.05). 3. Compared with control group, the expression ofα-SMA was signifi?cantly increased in high glucose group and S30 group (P<0.05). Compared with high glucose group, the expression of α-SMA was significantly decreased in S30 group (P < 0.05). Conclusion The 30 mmol/L GS can lead to TEMT in HK-2 cells. The more suitable inhibitory concentration of SB203580 in the process of TEMT is 30μmol/L. SB203580 can slow down the process of TEMT by inhibiting P38MAPK activation and inhibiting proliferation and the expression ofα-SAM s of HK-2 cells. |
| Author | 贾林 林智峰 马莉 唐玉玲 杨锐 杨晓萍 |
| AuthorAffiliation | 石河子大学第一附属医院肾病科,832008 |
| AuthorAffiliation_xml | – name: 石河子大学第一附属医院肾病科 邮编832008 |
| Author_FL | YANG Rui JIA Lin LIN Zhifeng MA Li TANG Yuling YANG Xiaoping |
| Author_FL_xml | – sequence: 1 fullname: JIA Lin – sequence: 2 fullname: LIN Zhifeng – sequence: 3 fullname: MA Li – sequence: 4 fullname: TANG Yuling – sequence: 5 fullname: YANG Rui – sequence: 6 fullname: YANG Xiaoping |
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| DocumentTitleAlternate | The effects of P38MAPK inhibitor SB203580 on TEMT of HK-2 cells |
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| Keywords | 糖尿病肾病 kidney tubules P38MAPK 上皮间质转分化 epithelial cells 上皮细胞 epithelial-mesenchymaltransition SB203580 P38丝裂原活化蛋白激酶 肾小管 diabetic nephropathy |
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| Notes | diabetic nephropathy; kidney tubules; epithelial cells; epithelial-mesenchymaltransition; P38MAPK; SB203580 JIA Lin, LIN Zhifeng, MA Li, TANG Yuling, YANG Rui, YANG Xiaoping （ Division of Nephrology, the First Affiliated Hospital, College of Medicine, Shihezi University, Shihezi 832008, China ） Objective To observe the effects of different concentrations of SB203580, the inhibitor of P38 MAPK, inprocess of high glucose（GS）-induced renal tubular epithelial-myofibroblast transdifferentiation（TEMT）. Methods Thecultured human renal tubular epithelial cells（HK-2） were divided into control group（5.5 mmol/L GS）, GS（30 mmol/L GS）group and different concentrations of SB203580（30 mmol/L GS ＋5, 10, 20 and 30 μmol/L SB203580） groups. The treat-ments were for 48 hours. MTT assay was used to observe cell proliferation. The median inhibiting concentration（IC50） was cal-culated. Western blot assay was used to detect the expressions of P38 MAPK, P-P38 MAPK and α-smooth muscle actin（α-SMA） in control group, high-glucose group an |
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| Snippet | 目的探讨不同浓度P38丝裂原活化蛋白激酶（P38MAPK）抑制剂SB203580在高糖诱导肾小管上皮细胞-肌成纤维细胞转分化（TEMT）过程中的机制及其较佳作用浓度。方法体外培养人近端肾小管上皮细胞（HK-2）并分为对照组（5.5 mmol/L GS）、DMSO组（5.5 mmol/L GS＋30μmol/L... R692; 目的：探讨不同浓度P38丝裂原活化蛋白激酶（P38MAPK）抑制剂SB203580在高糖诱导肾小管上皮细胞-肌成纤维细胞转分化（TEMT）过程中的机制及其较佳作用浓度。方法体外培养人近端肾小管上皮细胞（HK-2）并分为对照组（5.5 mmol/L GS）、DMSO组（5.5 mmol/L... |
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| SubjectTerms | P38丝裂原活化蛋白激酶 SB203580 上皮细胞 上皮间质转分化 糖尿病肾病 肾小管 |
| Title | P38MAPK抑制剂SB203580对高糖诱导HK-2细胞转分化的影响 |
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