LncRNA-Associated ceRNA Network in the Spinal Cord Reveals the Central Mechanism of Chronic Inflammatory Visceral Pain.

OBJECTIVE

To study and analyze the varieties across long non-coding RNAs (lncRNAs), microRNAs (miRNAs), and mRNAs in spinal cord, construct the competing endogenous RNAs (ceRNAs) network, and discuss the possible central mechanism in the development of chronic inflammatory visceral pain (CIVP).

METHODS

The colitis-associated CIVP rat model was prepared using 2,4,6-trinitrobenzene sulfonic acid (TNBS) enema. The abdominal withdrawal reflex (AWR), mechanical withdrawal threshold (MWT), and thermal withdrawal latency (TWL) were used to evaluate the rat's pain behaviors. The whole transcriptome sequencing technique was adopted to analyze the differentially expressed lncRNAs, miRNAs, and mRNAs in the spinal cord. LncRNA-associated ceRNA network was constructed and identified using the ceRNA MuTATE method and the hypergeometric distribution algorithm. The sequencing result was then verified using the quantitative real-time PCR (RT-qPCR).

RESULTS

Compared to the normal group, CIVP rat models had significantly increased AWR and decreased MWT and TWL (all P<0.05), suggesting hyperalgesia. In the spinal cord of CIVP rats, 158 lncRNAs, 24 miRNAs, and 1216 mRNAs were found to be differentially expressed. 3 top-scored lncRNAs, 1 pain-related lncRNA, 3 downstream miRNAs, and 2 mRNAs were selected for RT-qPCR verification, and 2 lncRNAs, 3 miRNAs, and 2 mRNAs were verified (all P<0.05).

CONCLUSION

LncRNA TCONS_00001477 and TCONS_00019815 may regulate Col2a1 and Hoxd10 mRNAs by competitively binding to miR-3120, miR-542-5p, and miR-450b-3p, thereby participating in colitis associated CIVP.