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Biological effect of ketamine in urothelial cell lines and global gene expression analysis in the bladders of ketamine‑injected mice.

Shen CH, Wang ST, Lee YR, Liu SY, Li YZ, Wu JD, Chen YJ, Liu YW - Mol Med Rep (2014)

Bottom Line: Ketamine arrested the cells in G1 phase and increased the sub‑G1 population, and also increased the barrier permeability of these cell lines.Global gene expression analysis of the animals' bladders following data screening identified ten upregulated genes and 36 downregulated genes induced by ketamine.A total of 52% of keratin family genes were downregulated, particularly keratin 6a, 13 and 14, which was confirmed by polymerase chain reaction analysis.

View Article: PubMed Central - PubMed

Affiliation: Department of Urology, Chiayi Christian Hospital, Chiayi 600, Taiwan, R.O.C.

ABSTRACT
Ketamine is used clinically for anesthesia but is also abused as a recreational drug. Previously, it has been established that ketamine‑induced bladder interstitial cystitis is a common syndrome in ketamine‑abusing individuals. As the mechanisms underlying ketamine‑induced cystitis have yet to be revealed, the present study investigated the effect of ketamine on human urothelial cell lines and utilized a ketamine‑injected mouse model to identify ketamine‑induced changes in gene expression in mice bladders. In the in vitro bladder cell line assay, ketamine induced cytotoxicity in a dose‑ and time‑dependent manner. Ketamine arrested the cells in G1 phase and increased the sub‑G1 population, and also increased the barrier permeability of these cell lines. In the ketamine‑injected mouse model, ketamine did not change the body weight and bladder histology of the animals at the dose of 30 mg/kg/day for 60 days. Global gene expression analysis of the animals' bladders following data screening identified ten upregulated genes and 36 downregulated genes induced by ketamine. A total of 52% of keratin family genes were downregulated, particularly keratin 6a, 13 and 14, which was confirmed by polymerase chain reaction analysis. Keratin 14 protein, one of the 36 ketamine‑induced downregulated genes, was also reduced in the ketamine‑treated mouse bladder, as determined by immunohistochemical analysis. This suggested that cytotoxicity and keratin gene downregulation may have a critical role in ketamine‑induced cystitis.

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Ketamine increases urothelium barrier permeability in SV-HUC1, RT4 and 5637 cells. The cells were treated with 0–4 mM ketamine for 24 or 48 h, then the upper and bottom chamber media were incubated with Alexa Fluor® 488 goat anti-mouse immunoglobulin G and analyzed by a fluorescence microplate reader. Data are presented as the mean ± standard deviation of three independent experiments. *P<0.05, **P<0.01, ***P<0.001, significant difference between the control and ketamine-treated cells.
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f2-mmr-11-02-0887: Ketamine increases urothelium barrier permeability in SV-HUC1, RT4 and 5637 cells. The cells were treated with 0–4 mM ketamine for 24 or 48 h, then the upper and bottom chamber media were incubated with Alexa Fluor® 488 goat anti-mouse immunoglobulin G and analyzed by a fluorescence microplate reader. Data are presented as the mean ± standard deviation of three independent experiments. *P<0.05, **P<0.01, ***P<0.001, significant difference between the control and ketamine-treated cells.

Mentions: Due to the cytotoxicity of ketamine (Fig. 1), it was hypothesized that ketamine may decrease epithelial barrier function. Therefore, the urothelial barrier permeability assay was employed. Following ketamine treatment for 24 and 48 h, the permeability of green fluorescence-labeled antibodies was increased dose-dependently in SV-HUC-1, RT4 and 5637 cells (Fig. 2). When comparing the cytotoxicity of ketamine and its enhancing effect on the barrier permeability, it was evident that the dose causing cytotoxicity accompanied barrier function loss. This suggested that the cytotoxic effect of ketamine may, at least in part, cause the loss of barrier function in ketamine-treated urotheliums.


Biological effect of ketamine in urothelial cell lines and global gene expression analysis in the bladders of ketamine‑injected mice.

Shen CH, Wang ST, Lee YR, Liu SY, Li YZ, Wu JD, Chen YJ, Liu YW - Mol Med Rep (2014)

Ketamine increases urothelium barrier permeability in SV-HUC1, RT4 and 5637 cells. The cells were treated with 0–4 mM ketamine for 24 or 48 h, then the upper and bottom chamber media were incubated with Alexa Fluor® 488 goat anti-mouse immunoglobulin G and analyzed by a fluorescence microplate reader. Data are presented as the mean ± standard deviation of three independent experiments. *P<0.05, **P<0.01, ***P<0.001, significant difference between the control and ketamine-treated cells.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4262485&req=5

f2-mmr-11-02-0887: Ketamine increases urothelium barrier permeability in SV-HUC1, RT4 and 5637 cells. The cells were treated with 0–4 mM ketamine for 24 or 48 h, then the upper and bottom chamber media were incubated with Alexa Fluor® 488 goat anti-mouse immunoglobulin G and analyzed by a fluorescence microplate reader. Data are presented as the mean ± standard deviation of three independent experiments. *P<0.05, **P<0.01, ***P<0.001, significant difference between the control and ketamine-treated cells.
Mentions: Due to the cytotoxicity of ketamine (Fig. 1), it was hypothesized that ketamine may decrease epithelial barrier function. Therefore, the urothelial barrier permeability assay was employed. Following ketamine treatment for 24 and 48 h, the permeability of green fluorescence-labeled antibodies was increased dose-dependently in SV-HUC-1, RT4 and 5637 cells (Fig. 2). When comparing the cytotoxicity of ketamine and its enhancing effect on the barrier permeability, it was evident that the dose causing cytotoxicity accompanied barrier function loss. This suggested that the cytotoxic effect of ketamine may, at least in part, cause the loss of barrier function in ketamine-treated urotheliums.

Bottom Line: Ketamine arrested the cells in G1 phase and increased the sub‑G1 population, and also increased the barrier permeability of these cell lines.Global gene expression analysis of the animals' bladders following data screening identified ten upregulated genes and 36 downregulated genes induced by ketamine.A total of 52% of keratin family genes were downregulated, particularly keratin 6a, 13 and 14, which was confirmed by polymerase chain reaction analysis.

View Article: PubMed Central - PubMed

Affiliation: Department of Urology, Chiayi Christian Hospital, Chiayi 600, Taiwan, R.O.C.

ABSTRACT
Ketamine is used clinically for anesthesia but is also abused as a recreational drug. Previously, it has been established that ketamine‑induced bladder interstitial cystitis is a common syndrome in ketamine‑abusing individuals. As the mechanisms underlying ketamine‑induced cystitis have yet to be revealed, the present study investigated the effect of ketamine on human urothelial cell lines and utilized a ketamine‑injected mouse model to identify ketamine‑induced changes in gene expression in mice bladders. In the in vitro bladder cell line assay, ketamine induced cytotoxicity in a dose‑ and time‑dependent manner. Ketamine arrested the cells in G1 phase and increased the sub‑G1 population, and also increased the barrier permeability of these cell lines. In the ketamine‑injected mouse model, ketamine did not change the body weight and bladder histology of the animals at the dose of 30 mg/kg/day for 60 days. Global gene expression analysis of the animals' bladders following data screening identified ten upregulated genes and 36 downregulated genes induced by ketamine. A total of 52% of keratin family genes were downregulated, particularly keratin 6a, 13 and 14, which was confirmed by polymerase chain reaction analysis. Keratin 14 protein, one of the 36 ketamine‑induced downregulated genes, was also reduced in the ketamine‑treated mouse bladder, as determined by immunohistochemical analysis. This suggested that cytotoxicity and keratin gene downregulation may have a critical role in ketamine‑induced cystitis.

Show MeSH
Related in: MedlinePlus