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Search articles by 'Yi-Xiong Chen'. Chen YX 1 ,. Weng ZH ,. Zhang SL. Affiliations 1 author 1. Share this article Share with email Share with twitter Share with linkedin Share with facebook. Abstract Aim To investigate whether Notch signaling is involved in liver fibrosis by regulating the activation of hepatic stellate cells HSCs. Methods Immunohistochemistry was used to detect the expression of Notch3 in fibrotic liver tissues of patients with chronic active hepatitis.
Results The expression of Notch3 was significantly up-regulated in fibrotic liver tissues of patients with chronic active hepatitis, but not detected in normal liver tissues. Free full text. World J Gastroenterol. Published online Mar PMID: Author information Article notes Copyright and License information Disclaimer. All rights reserved. This article has been cited by other articles in PMC.
Go to:. Patients and liver biopsy samples Liver tissue samples were obtained by biopsy from 11 patients with chronic active hepatitis 5 women and 6 men; median age 43 years, range years. Table 1 Primer sequences for TaqMan real-time reverse transcription polymerase chain reaction. Open in a separate window. Western blotting assay Cells were washed with PBS and lysed. Figure 1. Figure 2. Figure 3. Figure 4. Figure 5. Research frontiers This study was undertaken to investigate whether Notch signaling is activated in HSCs and sequentially contributed to liver fibrosis by regulating the activation of HSCs.
Innovations and breakthroughs This is the first study to characterize the role of Notch signaling in liver fibrosis. Peer review This is a very interesting study aimed at investigating the role of the Notch signaling pathway in liver fibrosis. Pathogenesis of liver fibrosis. Annu Rev Pathol.
Wells RG. The role of matrix stiffness in hepatic stellate cell activation and liver fibrosis. J Clin Gastroenterol. Bataller R, Brenner DA.
Hepatic stellate cells as a target for the treatment of liver fibrosis. Semin Liver Dis. Embryonic lethality in mice homozygous for a processing-deficient allele of Notch1.
Notch signaling: cell fate control and signal integration in development. Notch1 signaling in FIZZ1 induction of myofibroblast differentiation. Am J Pathol. Epithelial Notch signaling regulates interstitial fibrosis development in the kidneys of mice and humans.
J Clin Invest. Preventive effect of Notch signaling inhibition by a gamma-secretase inhibitor on peritoneal dialysis fluid-induced peritoneal fibrosis in rats. Arthritis Rheum. Biochim Biophys Acta. Inhibition of Notch signaling prevents experimental fibrosis and induces regression of established fibrosis.
However, the relationship between miR and Notch signaling pathway has not been studied, and the interaction in liver fibrosis has not been proposed. Therefore, to explore the correlation between miR and Notch signaling pathway in the process of liver fibrosis is the focus of this study.
Based on previous research progress, this experiment mainly explores the expression of miR in patients with liver fibrosis; explores the role of Notch3 in the occurrence and development of human liver fibrosis; finally explores the impact of miR on the occurrence and development of liver fibrosis and the correlation with Notch3 signaling pathway. In addition, 19 cases of normal liver tissues outside of hepatic hemangioma were used as control group.
Exclusion criteria: 1 Patients with hepatitis C and other viral hepatitis, autoimmune liver disease, alcoholic liver disease, liver cancer or other malignant tumors; 2 Patients with chronic infectious diseases tuberculosis, hepatic hydatid, syphilis, human immunodeficiency virus infection ; 3 Patients with comorbid diabetes, hypertension, coronary atherosclerotic heart disease, cerebrovascular disease; 4 Signed informed consent was not obtained from the patient; 5 Patients with incomplete clinical data.
This study was approved by the ethics committee of the College of Basic Medical Sciences, Guizhou Medical University, and written informed consent was obtained from all participants Ethics No.
Liver tissue sections were subjected to HE staining. Hepatocyte degeneration, necrosis, and inflammatory cell infiltration were observed, scored, and graded. Liver tissue sections were subjected to Masson staining. Liver tissue sections were subjected to Sirius red staining. Leica DM polarizing microscope was used to observe the amount and degree of collagen deposition in the hepatic portal area, and the fibrosis stage was scored.
After adherent growth, the cells were digested with 0. Effects of Notch3 signaling on fibrosis in LX-2 cells, the following groups were set up: Notch3 upregulated group rNotch3 group , Notch3 downregulated group si-Notch3 group , control group, and scramble siRNA group.
Recombinant Notch3 was used to induce Notch3 signaling and Notch3 short interference si RNA was used to downregulate Notch3 expression, respectively Table 1. To examine the effects of miR on cell biology via the Notch signaling pathway, mimics groups transfected with miR mimics ; negative control groups transfected with negative control mimics ; inhibitor groups transfected with miR inhibitor mimics ; blank control groups were set up Table 1.
The transfection efficiency was observed by fluorescence microscope after 24 h. The cells in each group were cultured for 48 h, treated with 0. After mixing, the cells were placed at room temperature in dark for 15 min the cells were resuspended for 3 times , and then placed in ice bath; the apoptosis rate was detected by flow cytometry. After 24 h of culture, a straight line was gently drawn at each empty position with a sterile gun head, washed twice with normal saline, and photographed for recording.
After 24 h of culture, the scratch and coincidence were observed under the microscope. SPSS Student's t test was used for statistical analysis. Masson and he staining results showed that the structure of hepatic lobules in normal liver tissue and non fibrosis liver tissue was complete, and the arrangement of hepatic cords was regular. The mild group showed swelling of hepatocytes and a small amount of focal necrosis.
In the moderate group, the liver tissue showed structural disorder of hepatic lobules, obvious round vacuoles in liver cells, steatosis, aggravation of necrosis, infiltration of inflammatory cells and proliferation of fibrous tissue.
In severe group, a large number of fibrous bands and some fibrous septa were formed. The results of Sirius red showed that the collagen fibers in the normal group were limited to the vascular wall of the portal area, and the red type I collagen fibers, a small amount of type II collagen fibers and green type III collagen fibers could be observed under the polarizing microscope. With the aggravation of liver fibrosis, collagen deposition was observed under polarizing microscope, including a large number of red type I collagen fibers, green type III collagen fibers, a small number of type II collagen fibers and light yellow type IV collagen fibers.
There were 11 normal liver tissues in this experiment, 17 patients with mild liver fibrosis, 26 patients with moderate liver fibrosis, and 20 patients with severe liver fibrosis; the HE staining pictures of liver fibrosis were shown in Fig.
Pathological staining results of different degrees of liver fibrosis and normal liver tissue. There were three different staining results, HE staining, Masson staining and Sirius red staining. The control group was the normal liver tissue outside the hepatic hemangioma. Normal group was liver tissue with chronic hepatitis but without liver fibrosis.
Mild group was chronic liver disease with mild liver fibrosis. The moderate group was chronic liver disease with moderate liver fibrosis. The severe group was the liver tissue with chronic liver disease and severe hepatic fibrosis.
The expression of miR increased in patients with liver fibrosis, and was correlated with the severity of liver fibrosis. Expression differences of miR, Notch3 and Jagged1 in different liver fibrosis tissues A Differential tissue miR expression in patients with liver fibrosis at different stages of progression. C , D Difference of protein expression of Notch3 and Jagged1 in liver fibrosis tissues of different degrees. With the aggravation of liver fibrosis, the expression of Notch3 and Jagged1 increased gradually.
Effect of Notch3 signaling on expression of profibrogenic factors. Recombinant Notch3 was used to induce Notch3 signaling and Notch3 short interference si RNA was used to downregulate Notch3 expression, respectively.
The mRNA and protein expressions of the receptor Jagged1 and the pro-fibrinogen were up-regulated after Notch3 up-regulation, and the mRNA and protein expressions of the receptor Jagged1 and the pro-fibrinogen were down-regulated after Notch3 down-regulation.
The effects of Notch3 signaling pathway on the expression of fibrogenic factors are shown in Fig. Effect of miR on proliferation and apoptosis of human hepatic stellate cells. A — C Effect of miR on proliferation of human hepatic stellate cells. Experimental groups transfected with miR mimics ; negative control groups transfected with negative control mimics ; inhibitor groups transfected with miR inhibitor mimics ; blank control groups were set up. The up-regulation of miR promoted the proliferation of human hepatic stellate cells.
Downregulation of miR can inhibit the proliferation of human hepatic stellate cells. D , E Effect of miR on apoptosis of human hepatic stellate cells. The lower right quadrant represents apoptotic cells. Up regulation of miR inhibits apoptosis of human hepatic stellate cells. The apoptosis rate of miR inhibitors group was The effect of miR on human hepatic stellate cell migration was examined by cell scratch assay and Transwell assay Fig.
Effect of miR on migration of human hepatic stellate cells. A , C The migration rate of cells in different groups was detected by scratch test; B , D Transwell assay was used to detect cell mobility. Up regulation of miR promotes migration of human hepatic stellate cells. The expression of miR in the miR mimics group was significantly increased, while that in the inhibitor group was significantly decreased, indicating that the up-regulation and down-regulation of miR were effective.
Effects of miR on Notch3 signaling pathway and expression of fibrogenic factors. Up regulation of miR promotes the expression of Notch3 signaling pathway and fibrogenic factor mRNA and protein. Liver fibrosis is the destruction of liver parenchymal cells and self repair of tissue caused by many factors, which is characterized by excessive production and deposition of extracellular matrix mainly composed of collagen.
It is the only way for various chronic liver diseases to develop into cirrhosis. When chronic liver injury occurs, resting hepatic stellate cells HSC become active, which is the central link of liver fibrosis 2. The occurrence and development of liver fibrosis is a multifactorial and multifaceted process, resulting from the imbalance of several related genes.
MiRNAs have been found to be involved in the development of many human diseases, and their roles in the process of liver fibrosis have gradually become a research focus Guo et al. Jensen et al. This suggests that by studying the differential expression of miRNAs during liver fibrosis, it will be helpful for understanding the mechanisms of the development and progression of liver fibrosis, as well as for finding novel molecular regulatory targets of liver fibrosis.
A large number of experiments have confirmed that miRNA is closely related to the occurrence and development of liver fibrosis 16 , 17 , Up regulation of miR-9a-5p can induce HSC proliferation, migration and activation All the above studies indicate that miRNA is closely related to the occurrence and development of liver fibrosis, and the regulation of HSC activation, proliferation, apoptosis and migration by miRNA is one of the mechanisms of the occurrence and development of liver fibrosis, which may be mediated by various signaling pathways.
The relationship between Notch3 signaling pathway and liver fibrosis has also been deeply studied. Zheng et al. This result was also confirmed by Chen et al. The results also confirmed that Notch3 had the same effect in human hepatic stellate cells. The results showed that the expression of miR was up-regulated.
This study was undertaken to investigate whether Notch signaling is activated in HSCs and sequentially contributed to liver fibrosis by regulating the activation of HSCs. This is the first study to characterize the role of Notch signaling in liver fibrosis. This finding indicated that Notch3 may contribute to liver fibrosis by regulating the activation of HSCs.
This is a very interesting study aimed at investigating the role of the Notch signaling pathway in liver fibrosis. The text is generally well written, with structured abstract and organized sections.
The manuscript has scientific value since it includes original information about a relevant topic. Advanced Search. This Article. Citation of this article. Notch3 regulates the activation of hepatic stellate cells.
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Brief Article Open Access. All rights reserved. World J Gastroenterol. Mar 28, ; 18 12 : Published online Mar 28, Patients and liver biopsy samples. Cell line and culture conditions. Immunohistochemistry and immunocytofluorescence analysis. Transfection of siRNA and plasmid. TaqMan quantitive reverse transcription polymerase chain reaction. Table 1 Primer sequences for TaqMan real-time reverse transcription polymerase chain reaction. Expression of Notch3 in fibrotic liver tissues of patients with chronic active hepatitis.
Figure 1 Immunohistochemical staining of Notch3 in liver tissues of patients with chronic active hepatitis and normal livers. A: Intense staining of Notch3 in fibrotic tissues of livers from patients with chronic active hepatitis; B: Negative control; C: Notch3 was not detected in normal liver tissues x Expression of Notch3 in hepatic stellate cell-T6 cells. Figure 2 Immunofluorescence staining analysis was performed to examine expression of Notch3 in hepatic stellate cell-T6 cells.
A: The green fluorescence represents the expression of Notch3; B: Blue one represents nucleus of hepatic stellate cell-T6 cells x
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