RNA Sequencing Reveals the Wound Repair Mechanism of Cuyuxunxi Prescription in Surgical Patients with Anal Fistulas

Abstract

Background: Anal fistula is one of the most common colorectal and perirectal diseases in the world. Cuyuxunxi (CYXX) prescription is an efficient herbal fumigant used to promote the surgical wound healing of anal fistulas.

Objectives: This study aimed to explore the underlying molecular mechanism of CYXX prescription on surgical wound healing of anal fistulas.

Methods: Ten patients with anal fistula were randomized into a control group or treatment group. The wound surface of patients in the control group was rinsed by normal saline, while that in the treatment group was rinsed by CYXX prescription. The wound tissues of patients with anal fistulas seven days after the surgery were collected for hematoxylin-eosin (HE) staining and RNA sequencing. The expressions of differentially expressed genes (DEGs) were validated by real-time quantitative PCR (RT-qPCR).

Results: HE staining showed that CYXX treatment reduced the infiltration of inflammatory cells. A total of 472 DEGs, including 141 up-regulated genes and 331 down-regulated genes, were identified. These genes were significantly related to skin development, xenobiotic stimulus, and inflammation. In addition, the consistency rate of RT-qPCR and sequencing results was 83.33%, which showed a high relative reliability of the sequencing results.

Conclusion: CYXX prescription could improve epidermis repair and reduce inflammatory responses.

Keywords: Cuyuxunxi prescription; RNA; RNA sequencing; anal fistula; inflammatory cells; wound repair.