Laser Capture Microdissection on Surgical Tissues to Identify Aberrant Gene Expression in Impaired Wound Healing in Type 2 Diabetes

Williams, Rachael; Castellano-Pelicena, Irene; Al-rikabi, Aaiad H.; Sikkink, Stephen; Baker, Richard P.; Riches-Suman, Kirsten; Thornton, M. Julie

doi:10.3791/62091

Cited by 1 publication

(1 citation statement)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Laser capture microdissection (LCM) is a microscopicguided system that can isolate specific cell types or regions of interest from tissue sections with a laser beam [21]. A variety of studies have indicated that LCM is powerful for isolating distinct cells and is helpful for the subsequent analysis of DNA, RNA, or protein profiles by sequencing or microarray [22][23][24]. With respect to the intestine, LCM has been used to obtain high-integrity RNA samples from freshly resected human intestinal tissue and to evaluate the gene expression profiles between crypts and villi of ileal epithelial cells [25,26].…”

Section: Introductionmentioning

confidence: 99%

circFLNA promotes intestinal injury during abdominal sepsis through Fas-mediated apoptosis pathway by sponging miR-766-3p

Shi

Zhang

et al. 2023

Inflamm. Res.

View full text Add to dashboard Cite

Background Intra-abdominal infections are the second most common cause of sepsis in the intensive care unit. Intestinal epithelial injury due to abdominal sepsis results in a variety of pathological changes, such as intestinal bacteria and toxins entering the blood, leading to persistent systemic inflammation and multiple organ dysfunction. The increased apoptosis of intestinal epithelial cells induced by sepsis further exacerbates the progression of sepsis. Although several studies have revealed that circRNAs are involved in intestinal epithelial injury in sepsis, few studies have identified the roles of circRNAs in intestinal epithelial apoptosis. Methods We used laser capture microdissection to obtain purified epithelial cells located in intestinal crypts from four patients with abdominal sepsis induced by intestinal perforation and four samples from age and sex-matched non-septic patients. Microarray analysis of circRNAs was conducted to assess differentially expressed circRNAs between patients with and without sepsis. Lastly, in vitro and in vivo assays were performed to study the mechanism of circFLNA in intestinal epithelial apoptosis during sepsis. Results circFLNA was upregulated in the intestinal epithelium after abdominal sepsis induced by intestinal perforation. Inhibition of miR-766-3p impaired si-circFLNA-mediated inhibition of apoptosis and inflammation factor levels in lipopolysaccharide (LPS)-treated HIEC-6 cells. circFLNA aggravated apoptosis and inflammation through the Fas-mediated apoptosis pathway in both LPS-treated HIEC-6 cells and a mouse cecal ligation and puncture model. Conclusion Our findings showed that circFLNA promotes intestinal injury in abdominal sepsis through the Fas-mediated apoptosis pathway by sponging miR-766-3p. The circFLNA/miR-766-3p/Fas axis has potential as a novel therapeutic target for treating intestinal injury in sepsis.

show abstract