Effect of Monoacylglycerol Lipase Inhibition on Intestinal Permeability of Rats With Severe Acute Pancreatitis

Abstract

Background: Endocannabinoid 2-arachidonoylglycerol (2-AG) is an anti-nociceptive lipid that is inactivated through cellular uptake and subsequent catabolism by monoacylglycerol lipase (MAGL). In this study, we investigated the effects of MAGL inhibition on intestinal permeability and explored the possible mechanism.Methods: A rat model of severe acute pancreatitis (SAP) was established. Rats were divided into three groups according to treatment. We analyzed intestinal permeability to fluorescein isothiocyanate-dextran and the levels of inflammatory factors interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) and 2-AG. Hematoxylin and eosin staining was used to assess histological tissue changes. In vivo intestinal permeability was evaluated by transmission electron microscopy. We obtained ileum tissues, extracted total RNA, and applied RNA-sequencing. Sequencing data were analyzed by bioinformatics.Results: Inflammatory factor levels were higher, while 2-AG levels were lower in the SAP group compared with the control group. Administration of JZL184 to rats with SAP increased the levels of 2-AG and lowered the levels of IL-6 and TNF-α. Notably, intestinal permeability was improved by JZL184 as demonstrated by fluorescein isothiocyanate-dextran measurement, hematoxylin and eosin staining, and transmission electron microscopy. RNA-sequencing showed significant transcriptional differences in SAP and JZL184 groups compared with the control group. KEGG analysis showed that the up- or downregulated genes in multiple comparison groups were enriched in two pathways, focal adhesion and PI3K-Akt signaling pathways. Differential alternative splicing (AS) genes, such as Myo9b, Lsp1, and Git2, have major functions in intestinal diseases. A total of 132 RNA-binding proteins (RBPs) were screened by crossing the identified abnormally expressed genes with the reported RBP genes. Among them, HNRNPDL coexpressed the most AS events as the main RBP.Conclusion: MAGL inhibition improved intestinal mucosal barrier injury in SAP rats and induced a large number of differentially expressed genes and alternative splicing events. HNRNPDL might play an important role in improving intestinal mucosal barrier injury by affecting alternative splicing events.

Single-Cell RNA Sequencing Reveals Roles of Fibroblasts During Intestinal Injury and Repair in Rats with Severe Acute Pancreatitis

“…In our previous research, numerous RNA-binding proteins (RBPs) were differentially expressed during SAP-induced intestinal injury and MAGL treatment, indicating they may have important functions. 5 Based on this, we evaluated intersections of DEGs with known RBPs in different cell clusters between CON and SAP groups, and between SAP and JZL184 groups. We observed numerous upregulated and downregulated RBP genes in cell clusters (such as C6 and C10) in our comparisons of different groups ( Figure S5A – D ).…”

“…The researchers involved in this study were blinded to specific details regarding the treatments administered to the animals. The protocol for establishing the SAP rat model and conducting the JZL184 treatment experiments followed the method outlined in our previous study (Wang et al, 2022). Twenty-four hours after the surgery, the SD rats were randomly divided into three groups: the Control group (CON), the SAP group (SAP), and the SAP group treated with JZL184 (JZL184).…”

“…Our previous study found that the MAGL inhibitor JZL184 protected against intestinal mucosal permeability damage in SAP rats. 5 However, it is still unclear how MAGL inhibitors affect the composition and function of cellular subsets in intestinal tissue during the process of intestinal mucosal barrier damage caused by AP. Due to the presence of intestinal epithelial cells and large numbers of infiltrating immune cells in the tissue, MAGL inhibitor treatment may have varying effects on different cell types during SAP-induced intestinal injury.…”

Dysregulated RNA editing of EIF2AK2 in polycystic ovary syndrome: clinical relevance and functional implications

“…It has also been implicated in several muscular dystrophies, including HNRNPDL-related muscular dystrophy with varying clinical and MRI phenotypes [ 94 , 95 ]. Additionally, HNRNPDL is found to be the most coexpressed RBP with alternative splicing events in severe acute pancreatitis [ 96 ]. Our findings further suggest the potential involvement of RBPs in the regulation of RNA editing in PCOS.…”

Update on pain management in acute pancreatitis

“…Results from the latest meta-analysis [45 && ] showed that acupuncture was effective in reducing pain intensity, the time to relief, abdominal distention, and the time to relief of distension compared with routine treatment of acute pancreatitis pain. Two studies [47,48] noted that acupuncture reduced intestinal permeability and the local inflammatory response. Taken together there is now sufficient evidence to state that acupuncture reduces the need for opioids, reduces inflammation and promotes gastrointestinal motility.…”