Lung Morphology Change Of Sidt2 Knockout Mice And Relevant Signal Route Research

Objective:By ultrastructure observation and relevant molecular research of Sidt2 knockout mice lung tissue,discuss the influence of Sidt2 gene on the lung morphology.Make analyses of signal route about Endoplasmic reticulum stress(ERS)and cell apoptosis.Methods:1、Sidt2 knockout mouse models were obtained by using LoxP-Flox-LoxP system,which based on the technologies of genetic recombination and gene targeting.Make model B129 male mice and female mice of Sidt2 gene body eliminated mated in the cage.Extract DNA from rat tail to identify the genotype.Select offspring Sidt2-/+ mice,mated in the cage,then select offspring Sidt2-/-and Sidt2-/+ male mice as experimental group and male Sidt2+/+ mice in the same nest as contrast.After confirming genotype by identification of DNA,RNA and Western,observed lung morphology by light microscope and electron microscope.2、Test the level of Bip and Caspase3 activity of Sidt2+/+ 、Sidt2-/-mice lung morphology by immunohistochemistry.3、Test ERS and apoptosis relevant molecular : BIP、p-eif-2α、ire1α、chop、Caspase3activity、Caspase3 by western.4、Make interfere of Sidt2 in non-small cell lung cancer cell A549,then test the level of BIP.Results:1、Validate successful of model construction through DNA,RNA and protein level. 2、Under light microscope : control rat alveolar wall was smooth,only a small amount of granulocyte infiltration.Model rat alveolar interval blood capillary dilate,hyperemia.Inflammatory cell were focal distributed in alveolar interval.Under electron microscopy:the control rat form rule lung morphology,Model rat alveolar epithelial cell were disruption,capillary hyperemia and thickening of basement membrane.3、The level of Bip and caspase3 activity of model rat is lower than control rat by immunohistochemistry.4、Tests find out Bip、p-eif2α、ire1α、Caspase 3 activity、Caspase 3 are lower in model rats than in control rats by western(P <0.05).On the contrary,CHOP is higher in Sidt2+/+rice(P <0.05).5、There is no significant difference of BIP content in interfered A549 cell to the contrast(P>0.05).Conclusion:1、Sidt2 knockout can affect the lung morphology,characterized by cell apoptosis,capillary hyperemia,thickening of basement membrane,and infiltrated of a large number of inflammatory cells,then lead to lung damage.2、Sidt2 knockout inhibit part route of ERS by inhibiting Bip、p-eif-2αand ire1α,hence block the progress of normal cell apoptosis.Instead,cells are progressed into necrosis period.However,the elaborate of the mechanism has not been known,it nedds to be researched.