摘要
目的:探讨温莪术对转化生长因子(TGF-β1)诱导的肾小管上皮细胞转分化的作用及机制。方法:通过体外培养肾小管上皮细胞(NRK-52E)及体外药物血清技术,运用MTT法检测不同浓度温莪术含药血清对细胞增殖的影响,倒置显微镜观察细胞形态学改变,细胞免疫荧光法检测各组肾小管上皮细胞α-平滑肌肌动蛋白(α-SMA)mRNA表达的变化,实时荧光定量PCR法检测细胞整合素连接激酶(ILK)mRNA表达的变化,ELISA法检测细胞上清液中胶原Ⅰ(Col-Ⅰ)的分泌水平。结果:TGF-β1可诱导NRK-52E向成纤维细胞转化,出现肥大、拉长,显现梭形,胞浆内α-SMA表达明显增多(P<0.05),ILKmRNA的表达、Col-Ⅰ的分泌水平升高(P<0.05)。温莪术呈浓度依赖性抑制转分化发生,减少形态变化,下调α-SMA、ILK、Col-Ⅰ表达(P<0.05)。结论:TGF-β1可诱导转分化发生,伴有α-SMA、ILK、Col-Ⅰ等表达升高,而温莪术可抑制TGF-β1诱导的转分化,减少α-SMA、ILK、Col-Ⅰ的表达水平,并且有浓度依赖性。
Objective ： To observe the effect and mechanism of Curcuma aromatic on tubular epithelial - myofibroblast transdifferentiation induced by TGF - β1. MethocL ：The morphological changes were observed by inverted phase contrast mi- croscope, the proliferation of NRK -52E under different concentrations of Curcuma aromatic were detected by MTI＇. The a- SMA protein was assessed by immunohistochemistry ; the expression of ILK mRNA was detected by real time PCR ; themyofibroblast transdifferentiation, showing hypertrophy of elongated and fusiform - shaped cells with the enhanced expres- sions of ot - SMA and ILK（P 〈0. 05） ,and increased secretion of Col - I （P 〈0. 05）. After adding Curcuma aromatic, the morphology changes were improved, with the expressions of ILKmRNA and c^- SMA protein inhibited（P 〈 0. 05 ）and the concentration of Col - I decreased （P 〈 0. 05）in a dose - dependent manner. Conclusion：TGF - 131 could induce the NRK - 52E transdifferentiationg in vitro with the increased levels of c~ - SMA protein, ILK mRNA and Col - I . And Curcuma aromatic could inhibit it in a dose -dependent manner.
出处
《中华中医药学刊》
CAS
2013年第8期1708-1711,I0009,共5页
Chinese Archives of Traditional Chinese Medicine
基金
浙江省中医药管理局资助项目(2009ZA015)
