Mechanism of exogenous nucleic acids and their precursors improving the repair of intestinal epithelium after gamma-irradiation in mice - PubMed (original) (raw)

Mechanism of exogenous nucleic acids and their precursors improving the repair of intestinal epithelium after gamma-irradiation in mice

Da-Xiang Cui et al. World J Gastroenterol. 2000 Oct.

Abstract

AIM:To clone expressed genes associated with repair of irradiation-damaged mice intestinal gland cells treated by small intestinal RNA, and to explore the molecular mechanism of exogenous nucleic acids improving repair of intestinal crypt.METHODS:The animal mode of test group and control group was established, forty-five mice being irradiated by gamma ray were treated with small intestinal RNA as test group, forty mice being irradiated by gamma ray were treated with physiological saline as control group,five mice without irradiation were used as normal control, their jejunal specimens were collected respectively at 6h, 12h,24h, 4d and 8d after irradiation. Then by using LD-PCR based on subtractive hybridization, these gene fragments differentially expressed between test group and control group were obtained, and then were cloned into T vectors as well as being sequenced. Obtained sequences were screened against. GeneBank, if being new sequences, they were submitted to GeneBank.RESULTS:Ninety clones were associated with repair of irradiation-damaged intestinal gland cells treated by intestinal RNA. These clones from test group of 6h, 12h, 24h, 4d and 8d were respectively 18, 22, 25, 13, 12. By screening against GeneBank, 18 of which were new sequences, the others were dramatically similar to the known sequences, mainly similar to hsp, Nmi,Dutt1, alkaline phosphatase, homeobox, anti-CEA ScFv antibody, arginine/serine kinase and BMP-4,repA. Eighteen gene fragments were new sequences,their accept numbers in GeneBank were respectively AF240164-AF240181.CONCLUSION:Ninety clones were obtained to be associated with repair of irradiation damaged mice intestinal gland cells treated by small intestinal RNA, which may be related to abnormal expression of genes and matched proteins of hsp, Nmi, Dutt1, Na, K-ATPase,alkalineph-osphatase, glkA, single stranded replicative centromeric gene as well as 18 new sequences.

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Figures

Figure 1

Figure 1

Electrophoresis result of production of LD-PCR. 1: Marker; 2, 6: Positive control; 3, 4, 5, 7, 8: Results of 6 h, 12 h, 24 h, 4 d and 8 d; 9: Negative control

Figure 2

Figure 2

Results of cloning and identification of production of PCR. 1-4, 6-9: Results of part products cut by Bstz1; 5: Marker Ninety of positive clones were obtained from test group, positive clones of 6 h, 12 h, 24 h, 4 d and 8 d in test group were respectively eighteen, twenty-two, twenty-five, thirteen, twelve.

Figure 3

Figure 3

Part results of hybridization with RNA. A: Test group; B: Control group. Dot blots confirmed that these genes were overexpressed in test group, and lower expressed in control group, that is, these genes were associated with repair of intestinal gland cells treated by RNA.

Figure 4

Figure 4

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