| Objective: To detect the cryptdin gene(mRNA) expression in the small intestines of mouse and observe the effect of rhubarb on the expression.To probe into the mechanism of rhubarb to enhance gut barrier.Method: (1) To detect the cryptdin gene(mRNA) expression in the small intestine of mouse. 10 adult Kunming mice were divided randomly into 2 groups with 5 mice in each group: group Crl and group Cr4. Duodenum, jejunum and ileum were removed from the mice killed by cervical dislocation and cleaned of their blood and dejecta with physiological saline. 40mg of each small intestine, weighed with electronic balance, was prepared and their respective total mRNA was extracted in accordance with the specification of E.Z.N.A. Total RNA Kit;RT-PCR (reverse transcription polymerase chain reaction) was performed in accordance with the specification of One Step RNA PCR Kit (AMV) and under the following steps: (1)50℃ for 30 minutes; (2)94℃ for 2 minutes ; (3)94 ℃ for 40 seconds ; (4)60℃ for 40 seconds; (5) 72℃ for 40 minutes; (Note: the sequence of steps (3) through (5) was repeated 30 times consecutively.)(6)72℃ for 5 minutes. The PCR products were put in 1.5% agarose gel for electrophoresis under a voltage of 90V. Their respective target bar contents were measured on the basis of the American gel image analysis system Bio-Rad PAC and under the reference of 100bp Ladder DNA Marker. PCR products were purified according to the specification of PCR products purification kit and the purified products were taken to Genecore Company for DNA sequence analysis. (2) To observe the effect of rhubarb on the cryptdin gene expression in the small intestines of mouse. 20 adult Kunming mice were divided randomly into 2 groups with 10 mice in each group: the experimental group and the control group. The mice in the experimental group were gavaged with 0.5ml of 10% rhubarb decoction every 6 hours, and those in the control, with an equal amount of physiological saline. After 24 hours, small intestines were removed from the mice killed by cervical dislocation and were cleaned of their blood and dejecta with physiological saline. 40mg of each small intestine(ileum), measured of their length and weighed with electronic balance, was prepared for the experimental steps as mentioned in (1).Results: (1)There was cryptdin gene(mRNA) expression in each section of the small intestines of normal mouse and there was nostatistic significance in the cryptdin-1 gene expression difference in each of the small intestines, with p>0.05 (F=0.9531, v1=2, v2=8). (2)There was statistic significance in the cryptdin-4 gene expression difference in each of the small intestines, with p<0.01 (F=35.661, v1=2, v2=8). A pah-comparison based on SNK (Student-Newman-Keuls miltiple range test) of the average cryptdin-4 gene expressions (duodenum: 213.551ng/5ul; jejunum: 291.189ng/5ul; ileum: 423.179ng/5ul) in each section of the small intestines indicated that there was statistic significance in the difference between the average expressions, with p<0.01 (q=7.436 in the comparison between ileum and jejunum; q=11.811 in the comparison between ileum and duodenum; q=4.374 in the comparison between jejunum and duodenum). (3)There was no statistic significance in the difference of the body weight (t=1.289), the small intestine length (t=0.081) and the small intestine weight (t=0.358) between the experimental group and the control group, with p>0.20 (v=18). (4)The mean(427.087ng/5ul) of cryptdin-1 gene expression in mouse small intestine of the experimental group was higher than that (409.223ng/5ul) of the control group,but there was no statistic significance between them(t= 1.567, 0.10.5).(6)There was no statistic significance in the correlation between the cryptdin-1 andcryptdin-4 gene expression.Conclusion: (1)There was cryptdin gene(mRNA) expression in each section...
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