灯盏乙素抑制热应激诱导猪肾小管上皮细胞凋亡研究

灯盏乙素（Scu）为灯盏花中提取的单一黄酮类化合物，具有清热解毒和保护细胞免受热应激损伤的作用。试验将不同浓度Suc培养的猪肾小管上皮细胞（LLC—PK1）在42℃下热应激1h，采用RNA干扰法沉默细胞HSP72基因表达，荧光定量PCR和Western blot检测细胞热休克蛋白（HSP）70、Bcl-2、Bax、细胞色素C（Cyt—c）和caspase-3裂解。结果表明，Scu显著增加细胞Bcl-2蛋白和HSP70蛋白水平，增加Bcl-2／Bax比值，抑制cyt—c的释放及pro—caspase-3裂解和caspase-3表达；但沉默HSPT0后，细胞Cyt—c释放和caspase-3表达均...

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Published in东北农业大学学报 Vol. 47; no. 11; pp. 59 - 65
Main Author 丛霞 李华涛 陈健伟 张东君 曹荣峰 田文儒
Format Journal Article
LanguageChinese
Published 青岛农业大学动物科技学院,山东青岛,266109%肥城市畜牧兽医局,山东肥城,271600 2016
Subjects
HSP70
LLC—PK1
凋亡
灯盏乙素
热应激
heat stress
LLC-PK1
HSP70
灯盏乙素
apoptosis
热应激
凋亡
scutellarin
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ISSN1005-9369

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Summary:灯盏乙素（Scu）为灯盏花中提取的单一黄酮类化合物，具有清热解毒和保护细胞免受热应激损伤的作用。试验将不同浓度Suc培养的猪肾小管上皮细胞（LLC—PK1）在42℃下热应激1h，采用RNA干扰法沉默细胞HSP72基因表达，荧光定量PCR和Western blot检测细胞热休克蛋白（HSP）70、Bcl-2、Bax、细胞色素C（Cyt—c）和caspase-3裂解。结果表明，Scu显著增加细胞Bcl-2蛋白和HSP70蛋白水平，增加Bcl-2／Bax比值，抑制cyt—c的释放及pro—caspase-3裂解和caspase-3表达；但沉默HSPT0后，细胞Cyt—c释放和caspase-3表达均显著增加。由此可知，Scu抑制LLC—PK1凋亡，保护细胞免受热应激损伤。
Bibliography:23-1391/S
Scutellarin （Scu） is known to abate fever in the clinical practice of Traditional Chinese Medicine. The primary cultured pig kidney proximal tubular （LLC-PK1） cells incubated with （or without） various concentrations of Scu were heat-stressed at 42 ℃ for 1 h to make heat shock injury cellular model. RNAi technique was used to silence heat shock protein （HSP） 70 mRNA expression. The HSP70, Bcl-2, Bax, cytochrome c （cyt-c） and pro-caspase-3 cleavage of the cells were all detected by using real-time PCR and Western blot aimed to examine the effects of Scu on heat-stressed LLC-PK1 cells and to explore its possible mechanism. The results showed that Scu significantly increased the levels of Bcl-2 and HSP70 protein, the ratio of Bcl-2 to Bax, and inhibited the release of cyt-c, pro-caspase-3 cleavage as well as caspase-3 expression. Interestingly, caspase-3 expression and release of cyt-c were dramatically increased in the cells with HSP70 RNA interference. These results demonstrate that Scu protects LLC-PK
ISSN:1005-9369