摘要
目的观察RNA干扰CD36基因表达对博莱霉素(BLM)诱导的大鼠肺泡巨噬细胞潜在转化生长因子-β1(L-TGF-β1)活化影响。方法实验设BLM组、Lv-shCD36(CD36 shRNA重组慢病毒)组、Lv-shCD36对照组,各组细胞均用BLM刺激,采用realtime-PCR和western blot检测细胞中CD36干扰效率;采用貂肺上皮细胞(CCL-64)增殖抑制实验检测培养上清中TGF-β1活性。结果 Lv-shCD36组细胞CD36 mRNA表达均低于BLM组和Lv-shCD36对照组,与Lv-shCD36对照组比较,干扰效率可达83%;与BLM组、Lv-shCD36对照组比较,Lv-shCD36组细胞CD36蛋白表达均降低;Lv-shCD36组培养上清中活化转化生长因子-β1(TGF-β1)含量(91.82±36.79)pg/10μL和TGF-β1活化率(19.80±7.98)%均低于BLM组和Lv-shCD36对照组(P<0.05)。结论通过RNA干扰CD36基因表达能够抑制BLM诱导的大鼠肺泡巨噬细胞分泌的L-TGF-β1的活化。
Objective To study the influence of CD36 gene suppression induced by RNA interference on transfor- ming growth factor-β1 （L-TGF-β1） activation derived from bleomycin-treated macrophages in rats. Methods The three groups including bleomycin（BLM） group,BLM ＋ Lv-shCD36（ a lentiviral vector expressing shRNA specific against rat CD36 ） group, and BLM ＋ Lv-shCD36-NC（ non-silence control lentivirus） group were set for the experiment. The cells in each group were incubated with bleomycin and then collected for detecting the interference effect with real-time PCR and western blot, and the supernatant was collected for detection of TGF-β1 by CCL-64 mink lung epithelial growth inhibition assay. Results The expression of CD36 mRNA in the Lv-shCD36 group was significantly lower than in the BLM group and control group. Compared to the control group, the efficiency of interference was 83 %. The expression of CD36 pro- tein in the Lv-shCD36 group was also lower than in the BLM group and control group. The quantity （91.82 ± 36. 79 pg/ 10μ L） and percent （ 19. 80 ± 7.98% ）of active TGF-β1 in the Lv-shCD36 group were significantly lower than those in the BLM group and control group（P 〈 0. 05 for all）. Conclusion Suppression of the CD36 expression by RNA interference could inhibit the activation of L-TGF-β1 derived from bleomycin-treated macrophages in rat.
出处
《中国公共卫生》
CAS
CSCD
北大核心
2013年第3期363-365,共3页
Chinese Journal of Public Health
基金
国家自然科学基金(81102106)
