Effects of Notch3 on gene expression and signal pathway of pancreatic stellate cell activation

doi:10.12047/j.cjap.6084.2021.024

Abstract

Abstract: Objective: The pancreatic stellate cells( PSCs) of mice were isolated and cultured and the effects of Notch3 siRNA on PSCs gene expression were detected. Methods: were PSCs of mice were isolated and cultured. The expressions of α-SMA, fibonectin and collagen I in activated PSCs were detected by immunofluorescence. The PSCs were divided into four groups, blank control group (MOCK group), negative siRNA control group (NC group) , Notch3 siRNA group (N3 siRNA group) and Notch3 siRNA-1 group (N3 siRNA-1 group). Cell treatment: the same transfection method was applied to transfect PSCs for 48h. Then total RNA was extracted from each group, and the concentration and purity of RNA was measured. The transcriptome sequencing and analysis were performed by ANOROAD Gene technology (Beijing) co., LTD. Results: The immunofluorescence results showed that α-SMA, fibonectin and collagen I were significantly expressed in activated PSCs. The analysis of the sequencing results showed that the gene expressions of α-SMA, collagen I, fibronectin, CTGF and PCNA of PSCs were down-regulated in N3 siRNA group and N3 siRNA-1 group comparing with NC group. The genes involved in collagen metabolism were up-regulated, the gene expression of positive regulation of collagen biosynthesis was down-regulated, while that of negative regulation of collagen biosynthesis was up-regulated in N3 siRNA group and N3 siRNA-1 group comparing with NC group. The genes that regulated cell aggregation were down-regulated, and the genes that regulated extracellular matrix were down regulated in N3 siRNA group and N3 siRNA-1 group comparing with NC group. Inhibition of Notch3 expression in PSCs could affect the gene expressions of cell adhesion molecule signaling pathway, MAPK signaling pathway and TGF-β signaling pathway. Conclusion: Inhibition of Notch3 expression can inhibit activation of PSCs , and reduce the ability of proliferation, migration and aggregation and ECM synthesis of PSCs. Inhibition of Notch3 expression may affect other signal pathways such as cell adhesion molecule signaling pathway, MAPK signaling pathway and TGF-β signaling pathway, but its effects need further validation.

Key words: pancreatic stellate cell, Notch3, gene expression, signaling pathway

CLC Number:

R332

SONG Hai-yan, ZHOU Zhi-xin, ZHANG Yu-xiang. Effects of Notch3 on gene expression and signal pathway of pancreatic stellate cell activation[J]. CJAP, 2021, 37(4): 349-353.

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