The Effects Of Mesenchymal Stem Cell Exosome With An Overexpression Of Mir-148a On Hepatic Ischemia-Reperfusion Injury

Purpose: This study aimed to investigate the effect of the overexpression of miR-148a in mesenchymal stem cell exosomes on hepatic ischemia-reperfusion (I/R). Methods: Fat MSCs were extracted from the fat tissue around the kidneys of rats. Lentivirus-infected MSC cells loaded with an miR-148a overexpression sequence or an unrelated sequence vector were used to extract the over-expressed sequence, the unrelated sequence, and the normal MSC exosome cells. The perihepatic ligament was broken in the sham operation group, and no other treatment was performed. The model group was not injected with the exosome solution. The experiment group was injected via the tail vein with transfected exosomes with miR-148a overexpression. The expressions of miR-148a, TLR4 mRNA, and CaMKII mRNA in the liver tissues were determined using RT-qPCR. All the rats' liver tissues were observed after the HE staining, and the apoptosis of the liver cells was tested using TUNEL. The expressions of the TLR4, CaMKII, Bcl-2, and Bax proteins in the liver tissues were determined using Western blot. ELISA was used to determine the expressions of IL-1 beta and TNF-alpha in the serum. The TargetScan prediction and dual-luciferase reporter system were used to identify the relationship between miR-148a and CaMKII. Results: The MSC and the exosomes were successfully extracted and identified. The miR-148a in the overexpression group was significantly higher than it was in the other groups (P<0.05). The TLR4, CaMKII mRNA, and protein in the overexpression group were significantly lower than they were in the model and experimental groups (P<0.05). The TargetScan and dual-luciferase reporter system confirmed the targeted regulation relationship between miR-148a and CaMKII. Conclusion: The overexpression of miR-148a in mesenchymal stem cell exosomes can inhibit the expressions of CaMKII and TLR4 in I/R tissues and reduce the occurrence of the inflammatory response and apoptosis.