Hepatic connective tissue growth factor expression and regulation differ between non-steatotic and non-alcoholic steatotic livers from brain-dead donor

Dong-Jing Yang,Shui-Jun Zhang,Mohammed Shakil Ahmed,Wen-Zhi Guo,Ji-Hua Shi,Zong-Ping Xia

doi:10.1038/s41598-021-83516-2

Abstract

Accurate evaluation of liver steatosis is required from brain-dead donors (BDDs) with nonalcoholic fatty liver disease (NAFLD). Our purposes were to investigate expression and regulation of connective tissue growth factor (CTGF) expression in livers from human and rat after brain death, and further evaluate its potential application. NAFLD and brain death models were established in rats. LX2 cells were cultured under hypoxia/reoxygenation. CTGF protein and mRNA levels were measured in liver samples from BDDs of human and rat by immunohistochemistry and reverse transcription-quantitative polymerase chain reaction. YAP-regulated CTGF expression was investigated in LX2 cells via YAP small interfering RNA and Verteporfin treatment. Blood CTGF level from BDDs was measured by enzyme-linked immunosorbent assay. After brain death, CTGF, transforming growth factor-β and YAP were overexpressed in non-alcoholic steatotic liver, whereas CTGF was downregulated in non-steatotic liver. Time-series analysis revealed that CTGF and YAP expression was comparable, as confirmed by inhibited YAP expression in LX2 cells. CTGF level and NAFLD activity were linearly correlated. CTGF expression and regulation differ between non-steatosis and nonalcoholic steatosis livers from BDDs. CTGF may be an important factor to evaluate graft quality from BDDs with NAFLD.

Highlights

Accurate evaluation of liver steatosis is required from brain-dead donors (BDDs) with nonalcoholic fatty liver disease (NAFLD)
In human cohort 1, intrahepatic connective tissue growth factor (CTGF) mRNA and protein expression were significantly down-regulated in the non-steatotic liver, whereas it was highly up-regulated in the non-alcoholic steatotic liver after brain death as determined by RT-qPCR (t = 2.483, P = 0.032, Fig. 1A) and IHC (t = 4.641, P = 0.001) (Fig. 1B,C)
We further explored the dynamic expression of CTGF in the non-steatotic liver from a welldescribed rat model of brain death

Summary

Introduction

Accurate evaluation of liver steatosis is required from brain-dead donors (BDDs) with nonalcoholic fatty liver disease (NAFLD). Our purposes were to investigate expression and regulation of connective tissue growth factor (CTGF) expression in livers from human and rat after brain death, and further evaluate its potential application. CTGF, transforming growth factor-β and YAP were overexpressed in non-alcoholic steatotic liver, whereas CTGF was downregulated in non-steatotic liver. CTGF expression and regulation differ between non-steatosis and nonalcoholic steatosis livers from BDDs. CTGF may be an important factor to evaluate graft quality from BDDs with NAFLD. Abbreviations BDD Brain-dead donor NAFLD Nonalcoholic fatty liver disease CTGF Connective tissue growth factor CCN Cellular Communication Network Factor TGF-β Transforming Growth Factor Beta YAP Yes Associated Protein NASH Non-alcoholic steatohepatitis ECM Extracellular matrix HSCs Hepatic stellate cells α-SMA Alpha smooth muscle actin. Few study has investigated the impact factors and the underlying mechanism of liver steatosis under acute stress of brain death before organ p rocurement[8]

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Conclusion