| Objective:To establish a method of RT-q PCR for detection of IL-17 of liver in mice infected with Echinococcus multilocularis (Em). To observe the expression of IL-17 in the liver of Balb/c mice infected by Em, further the mechanism of alveolar echinococcosis to be revealed. Methods:To establish the mouse model,100μl homogenized metacestode was injected into the anterior liver lobe of the experimental mice liver. After 4wk, liver was sampled to extract the total RNA, and cDNA was obtained by reverse transcription; Primer was designed based on the nucleotide sequence of IL-17 gene from GENE BANK (U43088.1) by using primer 5.β-actin as an inner control, the cDNA of mouse liver was constructed to detect the sensitivity and prepare the standard curve. The level of liver fibrosis and pathological alteration were observed by HE and Massion staining.The protein expression and location of IL-17 were analyzed by immuhistochemistry. Results:1) For RT-qPCR, the designed primer was specific for detecting IL-17 and the melt temperature was 81.0℃for each sample. The RT-qPCR assay had a detection limit of 1×102 pg mRNA, with a dynamic range of detection from 1×106 pg to 1×102 pg mRNA. The expression of IL-17mRNA increased significantly (t=-3.598, P<0.05) in the experimental group (0.008728±0.000984) compared to the control group (0.003823±0.00082) in 4 weeks.2) the inflammation and fibrosis were observed in the liver of infected group.3) IL-17 only observed in hepatic sinusoid in the control group, while in hepatic sinusoid, hyalomitome, cell membrane, fiber organization, focus germinal layer surrounding in infected group.The expression of IL-17increased significantly (t=-2.845, P<0.05) in the experimental group (2.64±0.66) compared to the control group (1.45±0.57). Conclusion:The established RT-qPCR assay for detecting IL-17 gene has high specificity and sensitivity. IL-17 may play an important role during Em infection.
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