The Experimental Study Of The Immunomodulatory Effect And Mechanism Of T Cell Ig And Mucin Domain Protein 3 In The Early Phase Of Acute Pancreatitis

The First Section: The value of T cell Ig and mucin domain protein 3 in the early phase of acute pancreatitisObjective Tim-3 has been thought to play a crucial immunomodulatory role in inflammatory response.No research is available on the potential role of soluble Tim-3(s Tim-3)in acute pancreatitis(AP).We conducted a prospective clinical study to characterize its role in the early phase of AP.Methods 44 AP patients(16 mild acute pancreatitis,28 None-mild acute pancreatitis)presented within 24 hours on admission and 20 healthy volunteers(NC)were included in our study.The expression of Tim-3 on monocyte was detected by flow cytometry.Serum IL-6,IL-10,TNF-?,and s Tim-3 was detected by ELISA.Results Expression of Tim-3 on monocyte and s Tim-3 were statistically significant differences in groups.The expression of Tim-3 on monocyte in the mild group were significantly increased compared with the none-mild group.The s Tim-3 level in the none-mild group were significantly increased compared with the mild group.The pro-inflammatory cytokines IL-6 and TNF-? levels and anti-inflammatory cytokines IL-10 level in the none-mild and mild groups were significantly elevated compared with the NC.The Tim-3 level was positively correlated with the IL-10.The s Tim-3 level was positively correlated with the IL-6 and TNF-?,but the s Tim-3 level displayed no obvious correlations with the IL-10 level.The s Tim-3 level was positively correlated with the APACHE ? score.Conclusions These results suggest that Tim-3 and s Tim-3 may participate in the early progression of AP by its immune regulation inflammatory cytokines.Measurement serum s Tim-3 may be as an early marker for predicting of AP.The Second Section: The experimental study of the immunomodulatory effect and mechanism of T cell Ig and mucin domain protein 3in the early phase of severe acute pancreatitis in mice Objectives The immunomodulatory function of inflammatory plays an important role in the occurrence and development of AP.Excessive inflammatory response dominates the pathological process in the early phase of AP.Tim-3 is generally considered as an ideal target for immunotherapy.It is important to know whether the Tim-3 pathway is also involved in the regulation of AP,and,if so,whether it plays a negative regulatory role.Methods SAP mice model were induced by intraperitoneal injection L-Arginin.Male BALB/c mice were randomly divided into the sham injection group(sham),the severe acute pancreatitis group(SAP),and the Tim-3 blockade group(Anti-Tim-3).Serum amylase activity and pancreatic myeloperoxidase(MPO)activity were assessed.Histopathological scores of the pancreas were calculated.The concentrations of serum IL-6,IL-10,TNF-? were evaluated by ELISA kits.Quantitative RT-PCR was performed to detect the transcript amounts of Tim-3,IL-6,IL-10,TNF-?,and Toll-like receptor 4(TLR4)in peritoneal macrophages.The levels of peritoneal macrophages Tim-3,TLR4,My D88 and NF-k B p65 were measured by western blot analysis.Results Tim-3 blockade aggravates pancreatic tissue injury.Compared to the sham group,IL-6,TNF-? and IL-10 were significantly elevated in the Tim-3 blockade group.Compared to the sham and SAP group,Tim-3,Tim-3 m RNA and protein in peritoneal macrophages were significantly decreased in the Tim-3 blockade group.Compared to the sham and SAP group,the expressions of pro-inflammatory cytokines IL-6 and TNF-? m RNA were all significantly upregulated in macrophage in Tim-3 blockade group.The expression of TLR4 m RNA was significantly increased in the Tim-3 blockade group compared to the SAP and sham group,respectively,and the expression of TLR4 m RNA was significantly increased in the SAP group compared to the sham group.Compared to the SAP group,Tim-3 blockade group significantly increased the TLR4,My D88 and p65 protein expressions,indicating that Tim-3 blockade promotes TLR4/My D88 and NF-?B activation.Conclusions The expression of TLR4,My D88 and p65 protein in TLR4/My D88-NF-?B signaling pathway may be negative regulated by Tim-3 on the surface of peritoneal macrophages,which may affect the inflammatory response of AP in mice.