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Increased expression of heat shock protein 72 protects renal proximal tubular cells from gentamicin-induced injury.

Wang Z, Liu L, Mei Q, Liu L, Ran Y, Zhang R - J. Korean Med. Sci. (2006)

Bottom Line: Both HSP72 protein and gene expression increased significantly at 72 hr when cells were treated with GM.It could reduce LDH release and NAG activity.HS also increased SOD activity, and decreased MDA content when cells were damaged by GM.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology, School of Pharmacy, Fourth Military Medical University, Xi'an, Shaanxi, 710032, China.

ABSTRACT
The nephrotoxicity of gentamicin (GM) has been widely recognized. Heat shock protein 72 (HSP72) has been reported to be a cytoprotectant. However, its cytoprotective effect against GM induced kidney injury has not yet been studied. In this study, we investigated the cytoprotective effect of HSP72 on GM-induced nephrotoxicity in vitro. Human Kidney tubular cell line, HK-2 cells were divided into four groups: control group, GM group (cells incubated with GM only), heat shock (HS) group (cells incubated at 43 degrees C for 30 min), and GM plus HS group, respectively. Lactate dehydrogenase (LDH) release increased time-dependently from 24 hr to 96 hr compared to the data of cells treated with GM only. Results of NAG activities, superoxide dismutase (SOD) activities and malondialdehyde (MDA) content were similar to that of the LDH release. The amount of HSP72 positive cells increased significantly at 72 hr after cells were treated with GM only. Both HSP72 protein and gene expression increased significantly at 72 hr when cells were treated with GM. On the other hand, HS induced HSP72 expression markedly. Pretreatment of HS inhibited HK-2 cells from GM-induced injury. It could reduce LDH release and NAG activity. HS also increased SOD activity, and decreased MDA content when cells were damaged by GM. These findings suggested that HS may protect kidney cells from GM-induced injury. Pre-induction of HSP72 may provide therapeutic strategies for nephrotoxicity induced by GM.

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Expression of HSP72mRNA in HK-2 cells. (A) RT-PCR analysis of HSP72 and GAPDH expression in HK-2 cells treated with medium only, HS, GM in 100 µg/mL and GM+HS, respectively, for 72 hr. (B) Quantification of HSP72 expression by densitometer. Values are mean±SD of measurements from separate HK-2 cell cultures. Number of experiments (n)=6. *p<0.01 vs. control, †p<0.01 vs. GM group at the same time point.
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Figure 8: Expression of HSP72mRNA in HK-2 cells. (A) RT-PCR analysis of HSP72 and GAPDH expression in HK-2 cells treated with medium only, HS, GM in 100 µg/mL and GM+HS, respectively, for 72 hr. (B) Quantification of HSP72 expression by densitometer. Values are mean±SD of measurements from separate HK-2 cell cultures. Number of experiments (n)=6. *p<0.01 vs. control, †p<0.01 vs. GM group at the same time point.

Mentions: After cells were heat shocked or exposed to GM at a concentration of 100 µg/mL for 72 hr, the rate of HSP72 positive cells increased dramatically (Fig. 6). It was higher than the control group at 72 hr after cells were exposed to GM+HS, but lower than the cells exposed only to GM. Densitometric measurements of HSP72 bands showed that the expression increased markedly after the exposure to GM or HS for 72 hr (Fig. 7). Proteins extracted from cells exposed to GM+HS showed that the expression of HSP72 increased than that of in control group, but was lower than GM group. Furthermore, from the results of RT-PCR, the change pattera of HSP72 pattern gene transcription showed similar result with the expression pattern of HSP72 (Fig. 8). An increased production of HSP72 gene is associated to a parallel rise of HSP72 protein content.


Increased expression of heat shock protein 72 protects renal proximal tubular cells from gentamicin-induced injury.

Wang Z, Liu L, Mei Q, Liu L, Ran Y, Zhang R - J. Korean Med. Sci. (2006)

Expression of HSP72mRNA in HK-2 cells. (A) RT-PCR analysis of HSP72 and GAPDH expression in HK-2 cells treated with medium only, HS, GM in 100 µg/mL and GM+HS, respectively, for 72 hr. (B) Quantification of HSP72 expression by densitometer. Values are mean±SD of measurements from separate HK-2 cell cultures. Number of experiments (n)=6. *p<0.01 vs. control, †p<0.01 vs. GM group at the same time point.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2722003&req=5

Figure 8: Expression of HSP72mRNA in HK-2 cells. (A) RT-PCR analysis of HSP72 and GAPDH expression in HK-2 cells treated with medium only, HS, GM in 100 µg/mL and GM+HS, respectively, for 72 hr. (B) Quantification of HSP72 expression by densitometer. Values are mean±SD of measurements from separate HK-2 cell cultures. Number of experiments (n)=6. *p<0.01 vs. control, †p<0.01 vs. GM group at the same time point.
Mentions: After cells were heat shocked or exposed to GM at a concentration of 100 µg/mL for 72 hr, the rate of HSP72 positive cells increased dramatically (Fig. 6). It was higher than the control group at 72 hr after cells were exposed to GM+HS, but lower than the cells exposed only to GM. Densitometric measurements of HSP72 bands showed that the expression increased markedly after the exposure to GM or HS for 72 hr (Fig. 7). Proteins extracted from cells exposed to GM+HS showed that the expression of HSP72 increased than that of in control group, but was lower than GM group. Furthermore, from the results of RT-PCR, the change pattera of HSP72 pattern gene transcription showed similar result with the expression pattern of HSP72 (Fig. 8). An increased production of HSP72 gene is associated to a parallel rise of HSP72 protein content.

Bottom Line: Both HSP72 protein and gene expression increased significantly at 72 hr when cells were treated with GM.It could reduce LDH release and NAG activity.HS also increased SOD activity, and decreased MDA content when cells were damaged by GM.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology, School of Pharmacy, Fourth Military Medical University, Xi'an, Shaanxi, 710032, China.

ABSTRACT
The nephrotoxicity of gentamicin (GM) has been widely recognized. Heat shock protein 72 (HSP72) has been reported to be a cytoprotectant. However, its cytoprotective effect against GM induced kidney injury has not yet been studied. In this study, we investigated the cytoprotective effect of HSP72 on GM-induced nephrotoxicity in vitro. Human Kidney tubular cell line, HK-2 cells were divided into four groups: control group, GM group (cells incubated with GM only), heat shock (HS) group (cells incubated at 43 degrees C for 30 min), and GM plus HS group, respectively. Lactate dehydrogenase (LDH) release increased time-dependently from 24 hr to 96 hr compared to the data of cells treated with GM only. Results of NAG activities, superoxide dismutase (SOD) activities and malondialdehyde (MDA) content were similar to that of the LDH release. The amount of HSP72 positive cells increased significantly at 72 hr after cells were treated with GM only. Both HSP72 protein and gene expression increased significantly at 72 hr when cells were treated with GM. On the other hand, HS induced HSP72 expression markedly. Pretreatment of HS inhibited HK-2 cells from GM-induced injury. It could reduce LDH release and NAG activity. HS also increased SOD activity, and decreased MDA content when cells were damaged by GM. These findings suggested that HS may protect kidney cells from GM-induced injury. Pre-induction of HSP72 may provide therapeutic strategies for nephrotoxicity induced by GM.

Show MeSH
Related in: MedlinePlus