葛根素对热应激诱导的LLC-PK1细胞HSP70表达的影响

摘要

为研究不同浓度葛根素对热应激诱导的LLC-PK1细胞HSP70 mRNA和蛋白表达的影响,筛选出葛根素诱导HSP70表达的最佳浓度。选取处于对数增长期的猪肾小管上皮细胞,分为37℃对照组和42℃热应激组,热应激组细胞先在加入不同浓度(0.01~100μmol/L不等)葛根素的培养基中培养,热应激之后,提取总RNA和总蛋白,用RT-PCR和Western Blot检测HSP70 mRNA和蛋白的相对表达量。结果显示,42℃热应激和不同浓度的葛根素均能显著诱导LLC-PK1细胞的HSP70 mRNA和蛋白的表达,并且,随葛根素浓度升高,HSP70表达量增加,在各浓度组中,10μmol/L的葛根素诱导HSP70的表达最显著(P<0.01)。上述结果证明,葛根素能够诱导热应激条件下LLC-PK1细胞 HSP70的表达,并且随葛根素浓度升高,HSP70表达量增加。%This experiment was to study the effect of puerarin on the thermal stress induced HSP70 expression in pig renal tubular epithelial cell (LLC-PK1),and to selected the optimal concentration of puerarin for inducing HSP70 expression. The LLC-PK1 in its logarithmic growth period was selected and divided into 37 ℃ control group and 42 ℃ heat stress group. The heat stress group of cell was incubated with various concentrations (from 0. 01-100 μmol/L ) of puerarin before the heat stress treatment,and then the total RNA and protein of the cells were ex-tracted and were detected by RT-PCR and Western Blot,respectively. The results showed that both thermal stress at 42 ℃ and puerarin with various concentrations significantly induced the HSP70 expression of LLC-PK1 (P<0. 01), and HSP70 expression increased as the puerarin concentration increased in the culture medium. The relative expres-sion of HSP70 in the heat stress group was the highest in the 10 μmol/L of puerarin group. The above results indi-cate that puerarin can induce HSP70 expression of LLC-PK1 under heat stress and this expressions increased as the puerarin concentration increased in the culture medium.