Mechanism Of Blocking Kv1.3 In Alleviating Acute Liver Injury & Effect Of Propofol On The Migration Of Macrophages

Acute liver injury is one of the common clinical problems and has received longterm attention from medical workers.The basic pathological process is massive damage of liver cells and decline of liver function.If the process of acute liver function injury is not controlled,it may lead to further increase in liver injury,resulting in delayed healing of the injury,the formation of chronic liver injury is cirrhosis;even worse,the degree of liver damage in the short term greatly exceeds the liver The compensatory ability leads to the occurrence of fulminant hepatic failure and seriously jeopardizes the life and health of the patient.The pathogenesis of acute liver injury has not yet been fully elucidated at this stage,but macrophages,as an important component of the body's immune system,participate in the entire process of injury and exert distinct biological effects in different stages.Kv1.3 is a voltage-gated potassium channel and many studies have demonstrated its important role in immune regulation.Previous studies in our laboratory found that a large number of Kv1.3 ion channels are also distributed on the macrophage membrane.Blocking Kv1.3 and/or affecting its expression will inevitably affect the function of macrophages,including the ability of the macrophages themselves to migrate.Therefore,the main scientific hypothesis of this study is that Kv1.3 blocks macrophage recruitment by inhibiting the ability of macrophage migration,thereby reducing acute liver injury.To verify the scientific hypothesis and further explore its possible mechanisms,we designed the following experiment:1.Establishment of LPS/D-GaIN induced acute liver injury model,while giving MgTX intervention to observe the damage changesThe acute liver injury model induced by LPS combined with D-GaIN was established successfully.The liver pathological and serological parameters of the model group were higher than those of the control group,and the difference was statistically significant.After the use of a specific blocker of Kv1.3,the liver injury index was reduced.Kv1.3 blocker alone had no adverse effects on the mice themselves.2.Detection of macrophage recruitment during acute liver injuryFor smears of mouse peritoneal macrophages and immunohistochemical staining of the liver with CD68,it was found that large numbers of peritoneal macrophages exuded in the L/D mice.At the same time,the proportion of intrahepatic macrophages increased.It suggests that a large number of monocyte-derived macrophages are recruited to the liver.However,after MgTX treatment,the peritoneal macrophages were reduced and the inflammatory macrophages recruited to the liver were correspondingly reduced.3.To explore the effect of MgTX on the migration ability of macrophages in vitroWhen treated with MgTX 10 nM,both scratch and transwell assays indicated that the migration ability of Raw264.7 cells was inhibited and the difference was statistically significant.4.Explore the effect of MgTX on the expression level of Kv1.3 in vitroAfter treatment with MgTX 10 nM,the Kv1.3 protein level and mRNA level of Raw264.7 cells did not change significantly,and the results of immunofluorescence also confirmed the above results.It suggests that the effect of MgTX on Kv1.3 is not achieved by affecting its expression level.5.Through silencing or overexpression of the biological function of Kv1.3To confirm the effect of Kv1.3 on macrophage migration function,we silenced and overexpressed Kv1.3 protein.The results showed that the macrophage migration ability decreased significantly after Kv1.3 was silenced.Overexpression of Kv1.3 resulted in increased macrophage migration.Our experiments show that Mtgx can inhibit macrophage migration and reduce its recruitment in the early stages of inflammation to reduce liver damage.