| Traumatic pancreatitis(TP)is a special type of acute pancreatitis caused by pancreatic injury.It is characterized by high latent and mortality.Because the early symptoms of traumatic pancreatitis are not obvious,it is usually difficult to detect and treat.In addition,traumatic pancreatitis often occurs with some severe complications,such as systemic inflammatory response syndrome(SIRS),shock,and multiple system organ failure,(MSOF).At present,the management of traumatic pancreatitis is mainly based on symptomatic supportive treatment to delay life,including intravascular administration,minimally invasive treatment,ultrasound or CT guided percutaneous puncture.Little research has been done on the repair of pancreatic injured tissue,the control of systemic inflammatory response,and the related molecular mechanisms after traumatic pancreatitis.Therefore,it has become an urgent problem to search for a safe,effective,and easy way to repair the traumatic pancreatic function.Human umbilical cord mesenchymal stem cells(hUC-MSCs)are a type of stem cells with multiple differentiation capacities and strong self-renewal capacity,which can differentiate into different cells under certain specific conditions,and hUC-MSCs can reduce the host immune response by inhibiting the proliferation of immune cells and interfering with the production and release of inflammatory factors.It is widely used and studied in the field of tissue engineering.In recent years,many scholars at home and abroad have considered that the advantage of hUC-MSCs may play a corresponding role by releasing human umbilical cord mesenchymal stem cells-derived exosomes(huc MSC-Ex)to act on target cells.Extensive Literature were confirmed that huc MSC-Ex has abundant space in tissue repair,control of inflammation and treatment of ischemic diseases,because huc MSC-Ex contain biologically active molecules such as nucleic acid,protein and lipid.Therefore,huc MSC-Ex would have high potential and clinical value in traumatic pancreatitis.In this study,we mainly explored repair effect of huc MSC-Ex on traumatic pancreatitis in rats.Methods and results:In the first part of the experiment: We obtained exosomes by subculture of hUC-MSCs,ultracentrifugation;transmission electron microscopy and Western blot were used to identify morphological and surface specific antigens of exosomes.And the size of huc MSC-Ex was analyzed by nanoparticle size analyzer.In the second part of the experiment: We selected 60 SD rats and randomly divided them into four groups of 15 rats,including a control group(Control group),a traumatic pancreatitis group(TP group),an human umbilical cord mesenchymal stem cells group(TP+ hUC-MSCs group),an human umbilical cord mesenchymal stem cells-derived exosomes group(TP + huc MSC-Ex group)to establish a model of traumatic pancreatitis in rats based on the area of injury.After intervention treatments using hUC-MSCs and huc MSC-Ex,the HE staining was used to evaluate the degree of pancreatic histopathological damage;the expression levels of amylase,lipase,pro-inflammatory,and anti-inflammatory factors were measured by ELISA;apoptosis was detected by TUNEL;the expression levels of apoptotic proteins Bax and Caspase-3 were determined by Western blot,the expression of apoptotic genes was explored by whole genome sequencing of pancreatic tissue transcriptome.Finally,the molecular mechanism of apoptosis in pancreatic acinar cells was detected by RT q PCR.The results indicated that huc MSC-Ex extraction from the supernatant of third-generation hUC-MSCs by ultracentrifugation could obtain the exosomes needed for the experiment based on the balance of economic cost and extraction purity.Compared with TP group,TP+hUC-MSCs group and TP+huc MSC-Ex group had significantly lower pancreatic histopathological scores(P < 0.05);the expression levels of serum amylase,lipase,IL-6,and TNF-α were significantly reduced(P < 0.05),whereas the expression levels of IL-10 and TGF-β were significantly higher(P < 0.05);the apoptotic index of TP group was significantly higher than control group(P < 0.05),while TP+hUC-MSCs group and TP+huc MSC-Ex group had significantly lower apoptotic index than TP group(P < 0.05),and the expression levels of apoptotic proteins Bax and Caspase-3 were significantly decreased in TP+hUC-MSCs group and TP+huc MSC-Ex group(P < 0.05).42% of genes were up-regulated in TP+huc MSC-Ex group.KEGG enrichment analysis found that the expression of genes related to apoptosis pathway was not detected in the up-regulated genes.Further,RT-q PCR indicated that the m RNA expression of Bax and Caspase-3 were inhibite in TP+huc MSC-Ex group.In conclusion,hUC-MSCs and huc MSC-Ex have the same effect on the treatment of traumatic pancreatitis in rats and can exert reparative and therapeutic effects on the injured pancreas by controlling the inflammatory response and inhibiting the apoptosis of pancreatic cells. |
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