目的 觀察不同劑量的龜板提取物對骨髓間充質(zhì)干細(xì)胞中分化抑制蛋白1（inhibitor of differentiation 1，Id1）的作用。方法 將構(gòu)建成功的PGL3-Id1啟動子用磷酸鈣共沉淀法轉(zhuǎn)染大鼠骨髓間充質(zhì)干細(xì)胞，龜板提取物分別作用于轉(zhuǎn)染后的骨髓間充質(zhì)干細(xì)胞12、24、36 h，選取作用最為明顯的36 h，每組分別加入0、1、3、30、100 μg/mL龜板提取物，藥物作用36 h后收集細(xì)胞分別應(yīng)用螢光素酶報告基因系統(tǒng)、RT-PCR和Western blotting檢測Id1的表達(dá)。結(jié)果 早期、小劑量龜板提取物對Id1的影響不大；隨著時間和劑量的增加，Id1的表達(dá)水平也逐漸升高。結(jié)論 龜板提取物促進(jìn)了骨髓間充質(zhì)干細(xì)胞中Id1的表達(dá)，劑量越大，作用時間越長，促進(jìn)作用越明顯。

Objective To observe the effect of different dosages of Plastrum Testudinis extracts (PTE) on the expression of inhibitor of differentiation 1 (Id1) in bone marrow derived mesenchymal stem cells (MSCs). Methods The PGL3-Id1 promoter was established and the MSCs were transfected with PGL3-Id1 promoter by calcium phosphate co-precipitation method. The transfected cells were treated with PTE for 12, 24, and 36 h, respectively. Different dosages (0, 1, 3, 30, and 100 μg/mL) of PTE were added into samples of 36 h group. After 36 h treatment, the cells were collected and luciferase activity measurement, RT-PCR, and Western blotting methods were used to detect the expression of Id1. Results The activity of short-term and low dose PTE was not substantial, along with the doses or time increasing, the expression of Id1 in bone marrow derived MSCs increased. Conclusion PTE increases the expression of Id1 in MSCs, and the activity is more significant with the dose and time increases.

國家自然科學(xué)基金資助項目（30772861）；肇慶市科技創(chuàng)新計劃項目（2013E181）