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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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Keratin 14 protein expression in mouse bladder tissues by immunohistochemical analysis. The slides of bladder tissue were hybridized with anti-keratin 14 antibodies and then photographed under ×400 microscopy. Upper and lower images, representative images from two different mouse bladders.
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f4-mmr-11-02-0887: Keratin 14 protein expression in mouse bladder tissues by immunohistochemical analysis. The slides of bladder tissue were hybridized with anti-keratin 14 antibodies and then photographed under ×400 microscopy. Upper and lower images, representative images from two different mouse bladders.

Mentions: Cytoskeletal keratins belong to intracellular intermediate filaments that connect to epithelial cell adhesion plaques in macula adherens and hemidesmosome sites. Numerous inherited skin-blistering diseases are caused by keratin gene mutations. There were 52 keratin family genes in the gene expression microarray chip. The majority of the keratins were downregulated by ketamine: 40% following 30 days and 52% following 60 days (Fig. 3A). The top ten downregulated keratins in the 60-day treatment are listed in Table III. The top three downregulated keratins, including 6a, 13 and 14 were confirmed by PCR analysis (Fig. 3B). Following deleting the genes with no significant difference (P>0.05), a heat map of residue keratin genes was constructed (Fig. 3C). Among the downregulated keratin genes, keratin 14 gene was among the top three genes following 30- and 60-day treatment. Keratin 14 was also among the selected top ten downregulated genes in Table II. To confirm the protein expression change of keratin 14, immunohistochemical analysis was applied. The results demonstrated that keratin 14 protein expression was also decreased in the 60-day murine urothelium (Fig. 4).


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)

Keratin 14 protein expression in mouse bladder tissues by immunohistochemical analysis. The slides of bladder tissue were hybridized with anti-keratin 14 antibodies and then photographed under ×400 microscopy. Upper and lower images, representative images from two different mouse bladders.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f4-mmr-11-02-0887: Keratin 14 protein expression in mouse bladder tissues by immunohistochemical analysis. The slides of bladder tissue were hybridized with anti-keratin 14 antibodies and then photographed under ×400 microscopy. Upper and lower images, representative images from two different mouse bladders.
Mentions: Cytoskeletal keratins belong to intracellular intermediate filaments that connect to epithelial cell adhesion plaques in macula adherens and hemidesmosome sites. Numerous inherited skin-blistering diseases are caused by keratin gene mutations. There were 52 keratin family genes in the gene expression microarray chip. The majority of the keratins were downregulated by ketamine: 40% following 30 days and 52% following 60 days (Fig. 3A). The top ten downregulated keratins in the 60-day treatment are listed in Table III. The top three downregulated keratins, including 6a, 13 and 14 were confirmed by PCR analysis (Fig. 3B). Following deleting the genes with no significant difference (P>0.05), a heat map of residue keratin genes was constructed (Fig. 3C). Among the downregulated keratin genes, keratin 14 gene was among the top three genes following 30- and 60-day treatment. Keratin 14 was also among the selected top ten downregulated genes in Table II. To confirm the protein expression change of keratin 14, immunohistochemical analysis was applied. The results demonstrated that keratin 14 protein expression was also decreased in the 60-day murine urothelium (Fig. 4).

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