2014Journal of Beijing University of Traditional Chinese MedicineRequires access

Influence of Puerarin Injection on expression of GRP78 mRNA in HK-2 cells induced by high glucose

Nie Yan-n

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Abstract

Objective To study the influences of Puerarin Injection on the proliferation of HK-2 cells and the expression of glucose regulated protein 78KD( GRP78) induced by high glucose. Methods The concentration gradient of puerarin was set as 5 g / L,10 g / L,20 g / L,40 g / L,80 g / L,160 g / L, 320 g / L and 640 g / L. The proliferation of HK-2 cells was detected by using methyl thiazolyl tetrazolium test( MTT). After passage and serum-free adhere-wall culturing for 24 h,HK-2 cells were divided into normal group,model group and treatment group( with Puerarin Injection) and were cultured continuously for 48 h. The total RNA were extracted from all groups and expression of GRP78 mRNA was detected by using real-time fluorescence quantitative polymerase chain reaction( RT-PCR). Results The inhibitory effect of Puerarin Injection on the proliferation of HK-2 cells was reinforced as the increase of Puerarin Injection's dose. After 48 h,the expression of GRP78 mRNA increased in model group compared with normal group( P 0. 01),and the expression increased in treatment group compared with model group ( P 0. 01). Conclusion Puerarin Injection has inhibitory effect on the proliferation of HK-2 cells.After high glucose stimulation for 48 h,ERS stress reaction will be induced,and Puerarin Injection can improve correct protein folding and recover intracellular homeostasis through increasing the expression of GRP78 mRNA.

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Objective To study the influences of Puerarin Injection on the proliferation of HK-2 cells and the expression of glucose regulated protein 78KD( GRP78) induced by high glucose. Methods The concentration gradient of puerarin was set as 5 g / L,10 g / L,20 g / L,40 g / L,80 g / L,160 g / L, 320 g / L and 640 g / L. The proliferation of HK-2 cells was detected by using methyl thiazolyl tetrazolium test( MTT). After passage and serum-free adhere-wall culturing for 24 h,HK-2 cells were divided into normal group,model group and treatment group( with Puerarin Injection) and were cultured continuously for 48 h. The total RNA were extracted from all groups and expression of GRP78 mRNA was detected by using real-time fluorescence quantitative polymerase chain reaction( RT-PCR). Results The inhibitory effect of Puerarin Injection on the proliferation of HK-2 cells was reinforced as the increase of Puerarin Injection's dose. After 48 h,the expression of GRP78 mRNA increased in model group compared with normal group( P 0. 01),and the expression increased in treatment group compared with model group ( P 0. 01). Conclusion Puerarin Injection has inhibitory effect on the proliferation of HK-2 cells.After high glucose stimulation for 48 h,ERS stress reaction will be induced,and Puerarin Injection can improve correct protein folding and recover intracellular homeostasis through increasing the expression of GRP78 mRNA.

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Available abstract

Objective To study the influences of Puerarin Injection on the proliferation of HK-2 cells and the expression of glucose regulated protein 78KD( GRP78) induced by high glucose. Methods The concentration gradient of puerarin was set as 5 g / L,10 g / L,20 g / L,40 g / L,80 g / L,160 g / L, 320 g / L and 640 g / L. The proliferation of HK-2 cells was detected by using methyl thiazolyl tetrazolium test( MTT). After passage and serum-free adhere-wall culturing for 24 h,HK-2 cells were divided into normal group,model group and treatment group( with Puerarin Injection) and were cultured continuously for 48 h. The total RNA were extracted from all groups and expression of GRP78 mRNA was detected by using real-time fluorescence quantitative polymerase chain reaction( RT-PCR). Results The inhibitory effect of Puerarin Injection on the proliferation of HK-2 cells was reinforced as the increase of Puerarin Injection's dose. After 48 h,the expression of GRP78 mRNA increased in model group compared with normal group( P 0. 01),and the expression increased in treatment group compared with model group ( P 0. 01). Conclusion Puerarin Injection has inhibitory effect on the proliferation of HK-2 cells.After high glucose stimulation for 48 h,ERS stress reaction will be induced,and Puerarin Injection can improve correct protein folding and recover intracellular homeostasis through increasing the expression of GRP78 mRNA.

Key concepts: Puerarin, Messenger RNA, Molecular biology, Chemistry, MTT assay, Real-time polymerase chain reaction, Intracellular, Stimulation

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