[The effect of fasudil via Rho/ROCK signaling pathway on the inflammation and fibrosis in human mesangial cells in high glucose medium].
[The effect of fasudil via Rho/ROCK signaling pathway on the inflammation and fibrosis in human mesangial cells in high glucose medium].
复制标题
法舒地尔通过Rho/ROCK信号通路对高糖培养基中人系膜细胞炎症和纤维化的影响
DOI：
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发表时间：
2011
期刊：
影响因子：
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通讯作者：
Yunquan Fu
中科院分区：
文献类型：
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作者：
D. Ma;Qiu;Xiao;Jing Li;Qing;Yunquan Fu
客观化
目的：研究法舒地尔对高糖诱导的人肾小球系膜细胞(HMCS)炎症和纤维化过程中Rho/ROCK信号通路的抑制作用。
方法
同步化培养的HMC分为：(1)正常葡萄糖对照组(NG，5.5mmoL/L葡萄糖)、(2)高糖组(hG，30mmoL/L葡萄糖)、(3)甘露醇组(MAN，5.5mmoL/L葡萄糖+24.5 mmoL/L甘露醇)、(4)高糖+法舒地尔组(hG+F，法舒地尔浓度分别为25、50、100µm o l/L)。分别于0、12、24、36、48、72h收集上清液和细胞，用实时荧光定量聚合酶链式反应方法检测细胞内RhoA、ROCK-I、结缔组织生长因子α的浓度变化，用ELISA法检测上清液中纤维连接蛋白、结缔组织生长因子、肿瘤坏死因子的蛋白含量。
结果
(1)高糖培养的人肾小球系膜细胞RhoA、ROCK-I和CTGF的表达明显高于正常对照组，并存在一定的时间依赖性。此外，MAN与NG比较，差异无统计学意义。(2)在高糖环境下，不同浓度的法舒地尔预处理和高糖培养24、48h后，HG+F细胞RhoA、ROCK-I、CTGF mRNA的表达均显著低于HG，且存在一定的浓度依赖关系。(3)高糖以时间依赖的方式促进系膜细胞分泌纤维连接蛋白、肿瘤坏死因子和肿瘤坏死因子α蛋白，而正常葡萄糖和甘露醇无此作用。(4)不同浓度的法舒地尔预处理和高糖培养12、24、36、48、72h后，FN、CTF3、α蛋白分泌较HG明显减少。
结论
法舒地尔通过抑制高糖激活的HMCS Rho/ROCK信号通路，减少下游炎症因子和细胞因子的分泌，减轻HMCS的炎症和纤维化。这为糖尿病肾病的治疗靶点提供了新的依据。
OBJECTIVE
To study the effect of fasudil on inhibiting the Rho/ROCK signaling pathway under high glucose in human mesangial cells (HMCs) inflammation and fibrosis.
METHODS
Synchronized HMCs were divided into following groups: (1) Normal glucose control group (NG, 5.5 mmol/L glucose); (2) High glucose group (HG, 30 mmol/L glucose); (3) Mannitol group (Man, 5.5 mmol/L glucose + 24.5 mmol/L mannitol); (4) High glucose + fasudil group (HG + F, the concentrations of fasudil were 25, 50 and 100 µmol/L, respectively). Collect the supernatant and cells at 0, 12, 24, 36, 48 and 72 h respectively, and determine the concentration changes of the RhoA, ROCK-I, connective tissue growth factor (CTGF)mRNA with real-time PCR method in the cells, then used the ELISA method to check the protein content of the fibronectin (FN), CTGF, TNFα in the supernatant.
RESULTS
(1) RhoA, ROCK-I and CTGF mRNA of the HMCs cultured under the high glucose expressed significantly higher than those in the normal group, and there was certain time-dependence. Besides, there was no statistic significance by comparing Man and NG. (2)Under the high glucose situation, after the fasudil pretreatment with different concentrations and 24 h or 48 h culture with high glucose, RhoA, ROCK-I, CTGF mRNA expression was significantly decreased in HG + F, compared with HG, and there was certain concentration-dependence. (3) High glucose increased the FN, CTGF, TNFα protein secretion of HMCs in a time-dependent manner, but normal glucose and mannitol had no such effect. (4) After the fasudil pretreatment with different concentrations and culture with high glucose for 12, 24, 36, 48, 72 h, the FN, CTGF, TNFα protein secretion was significantly reduced compared with HG.
CONCLUSION
Fasudil can reduce the secretion of downstream inflammatory factors and cytokines by inhibiting high glucose-activated HMCs Rho/ROCK signaling pathway, and reduce the inflammation and fibrosis of HMCs. This provides a new basis for the therapeutic target in the treatment of diabetic nephropathy.