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. 2014 Oct 8;7:702.
doi: 10.1186/1756-0500-7-702.

Independent Effects of Sham Laparotomy and Anesthesia on Hepatic microRNA Expression in Rats

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Free PMC article

Independent Effects of Sham Laparotomy and Anesthesia on Hepatic microRNA Expression in Rats

Wiebke Werner et al. BMC Res Notes. .
Free PMC article

Abstract

Background: Studies on liver regeneration following partial hepatectomy (PH) have identified several microRNAs (miRNAs) that show a regulated expression pattern. These studies involve major surgery to access the liver, which is known to have intrinsic effects on hepatic gene expression and may also affect miRNA screening results. We performed two-third PH or sham laparotomy (SL) in Wistar rats to investigate the effect of both procedures on miRNA expression in liver tissue and corresponding plasma samples by microarray and qRT-PCR analyses. As control groups, non-treated rats and rats undergoing anesthesia only were used.

Results: We found that 49 out of 323 miRNAs (15%) were significantly deregulated after PH in liver tissue 12 to 48 hours postoperatively (>20% change), while 45 miRNAs (14%) were deregulated following SL. Out of these miRNAs, 10 miRNAs were similarly deregulated after PH and SL, while one miRNA showed opposite regulation. In plasma, miRNA upregulation was observed for miR-133a and miR-133b following PH and SL, whereas miR-100 and miR-466c were similarly downregulated following anesthesia and surgery.

Conclusions: We show that miRNAs are indeed regulated by sham laparotomy and anesthesia in rats. These findings illustrate the critical need for finding appropriate control groups in experimental surgery.

Figures

Figure 1
Figure 1
miRNA expression in the liver of rats 24 hours after isoflurane anesthesia. The effect of anesthesia on miRNA expression in the liver was investigated by qRT-PCR using samples of untreated animals vs. samples taken 24 hours following 45 minutes of isoflurane anesthesia and pain medication. Relative miRNA expression ratios (normal liver was considered to be 1) were normalized against stably expressed U6 small nucleolar RNA (Student’s t-test, **p < 0.01, ***p < 0.001).
Figure 2
Figure 2
miRNA plasma levels in rats 24 hours after partial hepatectomy, sham operation, or isoflurane anesthesia. miRNA levels in plasma samples of Wistar rats 24 hours after partial hepatectomy (PH), sham laparotomy (SL), or anesthesia only (AN). Measurements were performed with qRT-PCR (n = 3). The results were normalized to untreated animals and are displayed in fold changes (miRNA levels in plasma samples from untreated animals was considered to be 1; Student’s t-test, *p < 0.05, **p < 0.01, ***p < 0.001). Statistically significant differences were not found 24 hours after the procedures between the PH- and SL-groups (Oneway anova, p > 0.05).

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