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Characterization of Changes in Global Genes Expression in the Distal Colon of Loperamide-Induced Constipation SD Rats in Response to the Laxative Effects of Liriope platyphylla.

Kim JE, Park SH, Kwak MH, Go J, Koh EK, Song SH, Sung JE, Lee HS, Hong JT, Hwang DY - PLoS ONE (2015)

Bottom Line: Among the transcripts up-regulated by constipation, 89 were significantly down-regulated and 22 were recovered to the normal levels by the AEtLP treatment.On the other hand, after the AEtLP treatment, ten of these genes down-regulated by constipation were up-regulated significantly and five were recovered to the normal levels.The major genes in the up-regulated categories included Serpina3n, Lcn2 and Slc5a8, whereas the major genes in the recovered categories were Tmem45a, Rerg and Rgc32.

View Article: PubMed Central - PubMed

Affiliation: Department of Biomaterials Science, College of Natural Resources & Life Science/Life and Industry Convergence Research Institute, Pusan National University, Miryang, 627-706, Korea.

ABSTRACT
To characterize the changes in global gene expression in the distal colon of constipated SD rats in response to the laxative effects of aqueous extracts of Liriope platyphylla (AEtLP), including isoflavone, saponin, oligosaccharide, succinic acid and hydroxyproline, the total RNA extracted from the distal colon of AEtLP-treated constipation rats was hybridized to oligonucleotide microarrays. The AEtLP treated rats showed an increase in the number of stools, mucosa thickness, flat luminal surface thickness, mucin secretion, and crypt number. Overall, compared to the controls, 581 genes were up-regulated and 216 genes were down-regulated by the constipation induced by loperamide in the constipated rats. After the AEtLP treatment, 67 genes were up-regulated and 421 genes were down-regulated. Among the transcripts up-regulated by constipation, 89 were significantly down-regulated and 22 were recovered to the normal levels by the AEtLP treatment. The major genes in the down-regulated categories included Slc9a5, klk10, Fgf15, and Alpi, whereas the major genes in the recovered categories were Cyp2b2, Ace, G6pc, and Setbp1. On the other hand, after the AEtLP treatment, ten of these genes down-regulated by constipation were up-regulated significantly and five were recovered to the normal levels. The major genes in the up-regulated categories included Serpina3n, Lcn2 and Slc5a8, whereas the major genes in the recovered categories were Tmem45a, Rerg and Rgc32. These results indicate that several gene functional groups and individual genes as constipation biomarkers respond to an AEtLP treatment in constipated model rats.

No MeSH data available.


Related in: MedlinePlus

Change in the protein level of FGF and ACE in the muscle and mucosa layer of constipation, vehicle- and AEtLP-treated constipation group.Total tissue lysates were prepared from each layer collected from the distal colons of loperamide injected rats treated with vehicle or AEtLP as described in the Materials and Methods. A total of 50 μg of protein per sample were immunoblotted with specific antibodies for each protein. Three samples were assayed in triplicate by Western blotting. Data are reported as the mean±SD. a, P<0.05 relative to the non-constipation group. b, P<0.05 relative to the vehicle-treated constipation group.
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pone.0129664.g004: Change in the protein level of FGF and ACE in the muscle and mucosa layer of constipation, vehicle- and AEtLP-treated constipation group.Total tissue lysates were prepared from each layer collected from the distal colons of loperamide injected rats treated with vehicle or AEtLP as described in the Materials and Methods. A total of 50 μg of protein per sample were immunoblotted with specific antibodies for each protein. Three samples were assayed in triplicate by Western blotting. Data are reported as the mean±SD. a, P<0.05 relative to the non-constipation group. b, P<0.05 relative to the vehicle-treated constipation group.

Mentions: To test the reliability of the results obtained from the microarray analysis, two up-regulated genes (Slc9a5, and Klk10) and two down-regulated genes (LCN2 and Atrip) were selected. When comparing the results of the microarray data and that of the RT-PCR, the pattern of increase was similar despite the difference in the increase or decrease rate (Fig 3). Furthermore, in the reliability test with western blot, the expression pattern of one down-regulated gene (Fgf) and one recovered gene (ACE) in muscle layer was consistent with the microarray data although the mucosa layer showed the different pattern (Fig 4). These results corroborate the data from microarray analysis. Especially, the protein expression from each layer indicated the possibility that the data from microarray may well reflect muscle layer than mucosa layer.


Characterization of Changes in Global Genes Expression in the Distal Colon of Loperamide-Induced Constipation SD Rats in Response to the Laxative Effects of Liriope platyphylla.

Kim JE, Park SH, Kwak MH, Go J, Koh EK, Song SH, Sung JE, Lee HS, Hong JT, Hwang DY - PLoS ONE (2015)

Change in the protein level of FGF and ACE in the muscle and mucosa layer of constipation, vehicle- and AEtLP-treated constipation group.Total tissue lysates were prepared from each layer collected from the distal colons of loperamide injected rats treated with vehicle or AEtLP as described in the Materials and Methods. A total of 50 μg of protein per sample were immunoblotted with specific antibodies for each protein. Three samples were assayed in triplicate by Western blotting. Data are reported as the mean±SD. a, P<0.05 relative to the non-constipation group. b, P<0.05 relative to the vehicle-treated constipation group.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4495015&req=5

pone.0129664.g004: Change in the protein level of FGF and ACE in the muscle and mucosa layer of constipation, vehicle- and AEtLP-treated constipation group.Total tissue lysates were prepared from each layer collected from the distal colons of loperamide injected rats treated with vehicle or AEtLP as described in the Materials and Methods. A total of 50 μg of protein per sample were immunoblotted with specific antibodies for each protein. Three samples were assayed in triplicate by Western blotting. Data are reported as the mean±SD. a, P<0.05 relative to the non-constipation group. b, P<0.05 relative to the vehicle-treated constipation group.
Mentions: To test the reliability of the results obtained from the microarray analysis, two up-regulated genes (Slc9a5, and Klk10) and two down-regulated genes (LCN2 and Atrip) were selected. When comparing the results of the microarray data and that of the RT-PCR, the pattern of increase was similar despite the difference in the increase or decrease rate (Fig 3). Furthermore, in the reliability test with western blot, the expression pattern of one down-regulated gene (Fgf) and one recovered gene (ACE) in muscle layer was consistent with the microarray data although the mucosa layer showed the different pattern (Fig 4). These results corroborate the data from microarray analysis. Especially, the protein expression from each layer indicated the possibility that the data from microarray may well reflect muscle layer than mucosa layer.

Bottom Line: Among the transcripts up-regulated by constipation, 89 were significantly down-regulated and 22 were recovered to the normal levels by the AEtLP treatment.On the other hand, after the AEtLP treatment, ten of these genes down-regulated by constipation were up-regulated significantly and five were recovered to the normal levels.The major genes in the up-regulated categories included Serpina3n, Lcn2 and Slc5a8, whereas the major genes in the recovered categories were Tmem45a, Rerg and Rgc32.

View Article: PubMed Central - PubMed

Affiliation: Department of Biomaterials Science, College of Natural Resources & Life Science/Life and Industry Convergence Research Institute, Pusan National University, Miryang, 627-706, Korea.

ABSTRACT
To characterize the changes in global gene expression in the distal colon of constipated SD rats in response to the laxative effects of aqueous extracts of Liriope platyphylla (AEtLP), including isoflavone, saponin, oligosaccharide, succinic acid and hydroxyproline, the total RNA extracted from the distal colon of AEtLP-treated constipation rats was hybridized to oligonucleotide microarrays. The AEtLP treated rats showed an increase in the number of stools, mucosa thickness, flat luminal surface thickness, mucin secretion, and crypt number. Overall, compared to the controls, 581 genes were up-regulated and 216 genes were down-regulated by the constipation induced by loperamide in the constipated rats. After the AEtLP treatment, 67 genes were up-regulated and 421 genes were down-regulated. Among the transcripts up-regulated by constipation, 89 were significantly down-regulated and 22 were recovered to the normal levels by the AEtLP treatment. The major genes in the down-regulated categories included Slc9a5, klk10, Fgf15, and Alpi, whereas the major genes in the recovered categories were Cyp2b2, Ace, G6pc, and Setbp1. On the other hand, after the AEtLP treatment, ten of these genes down-regulated by constipation were up-regulated significantly and five were recovered to the normal levels. The major genes in the up-regulated categories included Serpina3n, Lcn2 and Slc5a8, whereas the major genes in the recovered categories were Tmem45a, Rerg and Rgc32. These results indicate that several gene functional groups and individual genes as constipation biomarkers respond to an AEtLP treatment in constipated model rats.

No MeSH data available.


Related in: MedlinePlus