The effects of inhibition of the IP3R⁃Ca2 + pathway in APAP⁃induced liver inj ury and mitochondrial⁃associated endoplasmic reticulum membranes
10.19405/j.cnki.issn1000-1492.2023.07.003
- Author:
Wangting Xu
1
;
Yulin Song
1
Author Information
1. Dept of Gastroenterology, The First Afiliated Hospital of Anhui Medical University, The Key Laboratory of Digestive Disease of Anhui Province , Hefei 230022
- Publication Type:Journal Article
- Keywords:
acetaminophen;
liver injury;
2⁃aminoethoxydiphenylborate;
1 , 2 ⁃bis(2⁃aminophenoxy) ethane⁃N , N ,N ′ N ′⁃tetraacetic acid;
inositol 1 ,4 ,5 ⁃trisphosphate receptor;
calcium ion;
mitochondrial⁃associated endoplasmic re⁃ ticulum membranes
- From:
Acta Universitatis Medicinalis Anhui
2023;58(7):1077-1081
- CountryChina
- Language:Chinese
-
Abstract:
Objective : To explore the effects of inhibition of the inositol 1 , 4 , 5 ⁃trisphate receptor ( IP3R) ⅣCa2 + pathway on acetaminophen (APAP) Ⅳinduced liver injury in mice and its mitochondrial⁃associated endoplasmic reticulum membranes ( MAMs) .
Methods :40 SPF⁃rated male C57BL/6 mice were randomly divided into control group(n = 10) , model group( n = 10) , IP3R inhibitor(2⁃aminoethoxydiphenyl borate , 2 ⁃APB) group( n = 10) and the Ca2 + chelator [1 ,2 ⁃bis(2⁃aminophenoxy) ethane⁃N , N , N ′ N ′⁃tetraacetic acid , BAPTA⁃AM] group ( n = 10) . Mice in the model group ,2 ⁃APB group , and BAPTA⁃AM group were given a single intraperitoneal injection of APAP (300 mg/kg) . Half an hour before APAP injection , mice in the 2 ⁃APB group were injected intraperitoneally with 2 ⁃ APB(20 mg/kg) and mice in the BAPTA⁃AM group were injected intraperitoneally with BAPTA⁃AM(2. 5 mg/ kg) . The mice in each group were executed 24 hours after intraperitoneal injection of APAP. Serum alanine aminotransferase (ALT) and aspartate aminotransferase ( AST) were measured by chemical method. The pathological change of liver was observed by HE staining. The ultrastructural changes of mitochondria , endoplasmic reticulum and MAMs in liver cells were observed by transmission electron microscopy. The changes of Ca2 + in liver tissue homogenate were measured by calcium assay kit. The protein expression levels of mitofusin 1 ( MFN1 ) , mitofusin 2 (MFN2) , inositol 1 ,4 ,5 ⁃trisphate receptor(IP3R1) ,glucose regulated protein 75(GRP75) were detected by Western blot.
Results :Compared with the control group , the serum ALT and AST in the model group increased (P <0. 01) . Disorganized hepatocyte arrangement , hepatocyte degeneration and lobular central necrosis were observed. Breakage and disappearance of mitochondrial cristae , swelling and fracture of the endoplasmic reticulum and increase of the number of MAMs were also observed by transmission electron microscopy. The Ca2 + level in liver tissue homogenate increased ( P < 0. 01) . MFN1 and MFN2 protein expression decreased ( P < 0. 01) , IP3R1 and GRP75 protein expression increased( P < 0. 01) . Compared with the model group , the serum ALT and AST in 2 ⁃APB group and BAPTA⁃AM group decreased ( P < 0. 01 ) , the liver pathological changes were significantly improved , MAMs decreased , the content of Ca2 + in liver homogenate decreased(P < 0. 01) , MFN1 and MFN2 protein expression increased(P < 0. 01) , IP3R1 and GRP75 protein expression decreased(P < 0. 01) .
Conclusion :Inhibition of the IP3R⁃Ca2 + pathway protects against APAP⁃induced liver injury in mice. Its mechanism is related to reducing hepatic Ca2 + levels , reducing the number of MAMs , and regulating the expression of MAMs⁃related proteins.
- Full text:2024100916424644457抑制IP3R-Ca~(2+...线粒体内质网结构偶联的影响_徐王婷.pdf
