Pan Yi-ling, Li Bing, Ran Pi-xin
The State Key Laboratory of Respiratory Disease, the First Affiliated Hospital of Guangzhou Medical University, Guangzhou 510120, China.
Zhonghua Jie He He Hu Xi Za Zhi. 2013 Aug;36(8):581-6.
To investigate the effect of wood smoke condensate (WSC) on proliferation and necrosis of human airway smooth muscle cells (HASMCs).
Primary cultured HASMCs between passage 2 and 8 were divided into 3 groups: a control group, a WSC group and a cigarette smoke condensate (CSC) group. The viability of cells was examined by the CCK8 assays. The ratio of cellular proliferative stage (S phase) and cell cycle index were examined by fluorescent-labeled thymidine analogue uptake assays and flow cytometry. The expression of cyclin D1 was detected by quantitative reverse transcriptase polymerase chain reaction (Q-PCR) and Western blot. Cell apoptosis and necrosis were observed by the annexin-V and PI staining. Statistical analysis was performed by using the One-way ANOVA and LSD-t test.
Cell viability reached peak at WSC 1 mg/L[(126 ± 12)%] and at CSC 10 mg/L exposure level [(142 ± 11) %] respectively. While at WSC 10 mg/L and CSC 60 mg/L exposure levels, cell viability decreased significantly to 86% and 76%, respectively, as compared with that of the blank control group[(100 ± 0)%] (q = 3.63- 9.33, P < 0.05). In the WSC 1 mg/L group, the cell proliferation ratio and the expression of cyclin D1 protein were (124 ± 20)% and 1.31 ± 0.64, respectively, the differences being significant as compared with the blank control group [(100 ± 0)%, 1.0 ± 0.0] (q = 5.85, 5.91, P < 0.05), while the expression of cyclin D1 mRNA and the percentage of S+G2M phase were 1.18 ± 0.21 and (103 ± 4)%, respectively, not significantly different as compared to the control group [(100 ± 0)%, 1.0 ± 0.0], (q = 1.16, 2.05, P > 0.05). In the CSC 10 mg/L group, the above-mentioned values were (204 ± 45)%, 1.80 ± 0.25, (140 ± 6)%, 1.48 ± 0.2, respectively, which were significantly higher than those in the blank control group (q = 5.38-16.51, P < 0.05) and in the WSC group (q = 3.33-15.35, P < 0.05). However, when HASMCs were exposed to WSC 10 mg/L, the cell death ratio was (13.39 ± 0.15)%, higher than that of the blank control group [(1.57 ± 0.41)%] and the CSC group [(6.61 ± 1.91)%] (q = 18.03, 10.34, P < 0.05). Apoptosis ratio in the CSC 40 mg/L group was [(61.8 ± 10.6)%], higher than that of the blank control group [(0.0 ± 0.0)%] and the WSC group [(1.7 ± 0.4)%] (q = 17.44, 16.95, P < 0.05).
Exposure to WSC caused a weak proliferation of HASMCs, but resulted in cell necrosis instead of apoptosis at high doses. There was a slight difference in cell effects between the WSC group and the CSC group.
探讨木烟冷凝物（WSC）对人气道平滑肌细胞（HASMCs）增殖和坏死的影响。
将第2至8代原代培养的HASMCs分为3组：对照组、WSC组和香烟烟雾冷凝物（CSC）组。采用CCK8法检测细胞活力。通过荧光标记的胸腺嘧啶核苷类似物摄取试验和流式细胞术检测细胞增殖期（S期）比例和细胞周期指数。采用定量逆转录聚合酶链反应（Q-PCR）和蛋白质免疫印迹法检测细胞周期蛋白D1的表达。通过膜联蛋白V和碘化丙啶染色观察细胞凋亡和坏死情况。采用单因素方差分析和LSD-t检验进行统计学分析。
WSC 1 mg/L组[（126±12）%]和CSC 10 mg/L组[（142±11）%]细胞活力分别达到峰值。而在WSC 10 mg/L组和CSC 60 mg/L组，细胞活力与空白对照组[（100±0）%]相比显著降低，分别降至86%和76%（q = 3.63 - 9.33，P < 0.05）。WSC 1 mg/L组细胞增殖率和细胞周期蛋白D1蛋白表达分别为（124±20）%和1.31±0.64，与空白对照组[（100±0）%，1.0±0.0]相比差异有统计学意义（q = 5.85，5.91，P < 0.05），而细胞周期蛋白D1 mRNA表达和S + G2M期百分比分别为1.18±0.21和（103±4）%，与对照组[（100±0）%，1.0±0.0]相比差异无统计学意义（q = 1.16，2.05，P > 0.05）。CSC 10 mg/L组上述值分别为（204±45）%、1.80±0.25、（140±6）%、1.48±0.2，均显著高于空白对照组（q = 5.38 - 16.51，P < 0.05）和WSC组（q = 3.33 - 15.35，P < 0.05）。然而，当HASMCs暴露于WSC 10 mg/L时，细胞死亡率为（13.39±0.15）%，高于空白对照组[（1.57±0.41）%]和CSC组[（6.61±1.91）%]（q = 18.03，10.34，P < 0.05）。CSC 40 mg/L组细胞凋亡率为[（61.8±10.6）%]，高于空白对照组[（0.0±0.0）%]和WSC组[（1.7±0.4）%]（q = 17.44，16.95，P < 0.05）。
暴露于WSC可导致HASMCs轻度增殖，但高剂量时导致细胞坏死而非凋亡。WSC组和CSC组细胞效应存在细微差异。
Zotero 插件