本文選題：游離二氧化硅 + 肺泡上皮細(xì)胞　； 參考：《中國職業(yè)醫(yī)學(xué)》2017年02期

【摘要】：目的觀察游離二氧化硅(SiO_2)直接或間接作用于大鼠肺泡Ⅱ型上皮細(xì)胞株RLE-6TN而誘導(dǎo)發(fā)生上皮間質(zhì)轉(zhuǎn)化(EMT)的情況,及其對RLE-6TN表面標(biāo)志蛋白表達(dá)影響情況。方法 (1)采用支氣管肺泡灌洗術(shù)提取無特定病原體級SD大鼠肺泡巨噬細(xì)胞(AM)。以質(zhì)量濃度為0~140 mg/L的SiO_2混懸液處理AM和RLE-6TN并培養(yǎng)24、48和72 h后,采用CCK-8實(shí)驗(yàn)檢測細(xì)胞存活率,篩選后續(xù)實(shí)驗(yàn)染塵劑量。(2)取AM接種于Transwell的小室層與RLE-6TN共培養(yǎng)。設(shè)直接對照組(單獨(dú)RLE-6TN,不作染塵處理)、直接染塵組(單獨(dú)RLE-6TN,予質(zhì)量濃度為100 mg/L的SiO_2混懸液處理)、間接對照組(AM與RLE-6TN共培養(yǎng),不作染塵處理)和間接染塵組(AM與RLE-6TN共培養(yǎng),予質(zhì)量濃度為100 mg/L的SiO_2混懸液處理AM),分別于染塵0、24、48和72 h后收獲細(xì)胞,用免疫印跡法測定RLE-6TN中的E-黏鈣蛋白(E-cad)和α-平滑肌肌動蛋白(α-SMA)的相對表達(dá)水平。結(jié)果 (1)根據(jù)CCK-8結(jié)果,后續(xù)實(shí)驗(yàn)的SiO_2染塵劑量設(shè)為100 mg/L。(2)RLE-6TN中E-cad和α-SMA相對表達(dá)水平在染塵處理和染塵時間的交互效應(yīng)上均有統(tǒng)計(jì)學(xué)意義(P0.01)。其中,直接染塵組和間接染塵組RLE-6TN在48和72 h的E-cad相對表達(dá)水平均低于同時間點(diǎn)直接對照組(P0.05),直接染塵組RLE-6TN在72 h時間點(diǎn)的E-cad相對表達(dá)水平分別低于同組0和24 h時間點(diǎn)(P0.05),間接染塵組RLE-6TN在48和72 h時間點(diǎn)的E-cad相對表達(dá)水平均低于同組0 h時間點(diǎn)(P0.05);在48和72 h時間點(diǎn),直接染塵組和間接染塵組α-SMA相對表達(dá)水平分別高于同時間點(diǎn)的相應(yīng)對照組(P0.05),且間接染塵組RLE-6TN中α-SMA相對表達(dá)水平均高于直接染塵組(P0.05);2個染塵組RLE-6TN中α-SMA相對表達(dá)水平均呈隨著染塵時間的增加而增加的時間-效應(yīng)關(guān)系(P0.05)。結(jié)論游離SiO_2直接或間接染塵均可誘導(dǎo)RLE-6TN發(fā)生EMT,導(dǎo)致E-cad表達(dá)下調(diào),α-SMA表達(dá)呈現(xiàn)時間-效應(yīng)性上調(diào);間接染塵可能更易于誘導(dǎo)RLE-6TN發(fā)生EMT。
[Abstract]:Objective to observe the effect of free SiO-2) on the induction of epithelial mesenchymal transformation (EMTT) in rat alveolar epithelial cell line RLE-6TN directly or indirectly and its effect on the expression of RLE-6TN surface marker protein. Methods 1) the alveolar macrophages of SD rats were isolated by bronchoalveolar lavage. AM and RLE-6TN were treated with SiO_2 suspension of 0 ~ 140 mg/L and cultured for 24 h and 72 h. The survival rate of cells was detected by CCK-8 assay. The cell viability was detected by CCK-8 assay. The cells were inoculated in the compartment layer of Transwell and co-cultured with RLE-6TN. Direct control group (RLE-6TNN alone, without dust treatment, direct dust-exposed group (RLE-6TNN alone, treated with 100 mg/L SiO_2 suspension), indirect control group (RLE-6TNN) co-cultured with RLE-6TN and without Dust treatment) and indirect dust-exposed group (RLE-6TNN) were co-cultured with RLE-6TN. The cells were harvested after treatment with 100 mg/L SiO_2 suspension for 48 h and 72 h, respectively. The relative expression levels of E-cadand and 偽 -SMAin in RLE-6TN were determined by Western blot. Results (1) according to the CCK-8 results, the relative expression of E-cad and 偽 -SMA in the following experiment was set as 100 mg/L.(2)RLE-6TN. There was significant difference in the interaction between the time of dust exposure and the treatment time of E-cad and 偽 -SMA (P 0.01). Of which, The relative expression level of RLE-6TN in direct exposure group and indirect exposure group at 48 h and 72 h was lower than that in direct control group at the same time point (P0.05). The relative expression level of RLE-6TN in direct dust-exposed group at 72 h was lower than that in the same group at 0 h and 24 h, respectively. The relative expression level of RLE-6TN in the indirect dust-exposed group at 48 and 72 h was lower than that in the same group at 0 h and 72 h, and at 48 h and 72 h, the relative expression level of E-cad was lower than that in the same group at 0 h and 72 h, respectively. The relative expression level of 偽 -SMA in direct dust group and indirect dust group was higher than that in control group at the same time point, and the relative expression level of 偽 -SMA in RLE-6TN of direct dust group was higher than that of direct dust group, and the relative expression level of 偽 -SMA in RLE-6TN of two dust-exposed groups was higher than that of direct dust group. The relationship between time and effect increased with the increase of dust exposure time (P 0.05). Conclusion both direct and indirect exposure to free SiO_2 can induce the occurrence of EMTs in RLE-6TN, leading to down-regulation of E-cad expression and a time-dependent up-regulation of 偽 -SMA expression, and indirect dust exposure may be more likely to induce EMT-induced RLE-6TN.
【作者單位】： 鄭州大學(xué)公共衛(wèi)生學(xué)院;鄭州大學(xué)第三附屬醫(yī)院超聲科;
【基金】：國家自然科學(xué)基金(81472954)
【分類號】：R135.2

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