The aim of this research is to evaluate the effect of tetramethylpyrazine (TMP) and connective tissue growth factor (CTGF) miRNA plasmids on the expressive levels of CTGF, transforming growth factor-beta (TGF-beta) and type Ⅰ collagen of rat hepatic stellate cells (HSC) which are stimulated by high glucose. The rat HSCs which were successfully transfected rat CTGF miRNA plasmids and the rat HSCs which were successfully transfected negative plasmids were cultured in vitro. After stimulus of the TMP and the high glucose, the protein levels and gene expressive levels of CTGF, TGF-beta and type Ⅰ collagen were tested. The results indicated that high glucose increased the expression of CTGF mRNA, CTGF protein, TGF-beta mRNA,TGF-beta protein and type Ⅰ collagen (P<0.05). The expressive levels of CTGF mRNA, CTGF protein, TGF-beta mRNA, TGF-beta and type Ⅰ collagen in TMP group were lower than those in high glucose group and showed statistically significant differences (P0.05). Compared with high glucose group, the expressive levels of CTGF mRNA, CTGF protein, TGF-beta mRNA, TGF-beta and type Ⅰ collagen in rat CTGF miRNA plasmid interference group were significantly lower (P<0.05). However, no statistically significant difference was found in CTGF mRNA and CTGF protein levels between TMP group and CTGF miRNA group (P>0.05), while type Ⅰ collagen levels showed statistically significant differences (P<0.05). It is concluded that high glucose could promote the expressions of CTGF, TGF-beta and type Ⅰ collagen, and TMP and rat CTGF miRNA plasmids could reduce the expressions of CTGF, TGF-beta, type Ⅰ collagen.
ObjectiveTo observe effect of echinococcus cyst fluid on proliferation and cell cycle progression of rat hepatic stellate cells (HSC-T6), and to preliminarily reveal a new mechanisms of pro-fibrogenic effect of alveolar echinococcosis. MethodsHSC-T6 cells were treated with different concentrations (0.00-0.90 mg/mL) of echinococcus cyst fluid. Then, the morphological changes were observed under the inverted microscope, the impacts on proliferation and cell cycle were tested by the CCK-8 assay and flow cytometry respectively. ResultsAfter treated by echinococcus cyst fluids with different concentrations (0.00-0.90 mg/mL) for 24 h, the most cells shrinked fusiform shape with more slender synapses, and the proliferation activities were increased with the concentration of echinococcus cyst fluid increasing when it was higher than 0.05 mg/mL (P<0.05), the proportion of G0/G1 was decreased (P<0.05) and those of S and G2/M were increased (P<0.05) with the concentration of echinococcus cyst fluid increasing. ConclusionsEchinococcus cyst fluid could promote proliferation of HSC-T6 cells in a dose-dependent manner, which might due to its impact on cell cycle progression. Therefore, alveolar echinococcosis might promote hepatic fibrosis through regulating hepatic stellate cells, but further research on detail needs to be done in future.
The contractile force of hepatic stellate cells plays a very important role in liver damage, hepatitis and fibrosis. In this paper, a method based on polydimethylsiloxane (PDMS) thin micropillar arrays is proposed to measure the contractile force of human hepatic stellate cell line LX-2, which enables dynamic measurement of the subcellular distribution of force magnitude and direction. First, thin micropillar arrays on glass bottom dish were fabricated using two-step casting process in order to meet the working distance requirement of 100× objective lens. After hydrophilic treatment and protein imprint, cells were seeded on the micropillar arrays. LX-2 cells, which were quiesced by growth in serum-free medium, were activated by adding fetal bovine serum (FBS). The deflections of the micropillars were achieved by image processing technique, and then the contractile force of cells exerted on the micropillars was calculated according to mechanical simulation results, and was analyzed under both quiescent and activated conditions. The experimental results show that the average traction force of quiescent cells is about 20 nN, while the contractile force of activated cells increased to 110 nN upon adding FBS. This method can quantify the contractile force of LX-2 cell on subcellular scale in both quiescent and activated states, which may benefit pathology study and drug screen for chronic liver diseases resulted from liver fibrosis.
ObjectiveTo summarize the research progress on the role of exosomes derived from different sources in hepatic stellate cells.MethodThe experimental studies and clinical applications of exosomes from different cell sources effected on hepatic stellate cells were reviewed.ResultsIn the occurrence and development of liver fibrosis pathological physiological process, the activation, proliferation, migration, apoptosis of hepatic stellate cells played the important roles on the development of liver fibrosis. In recent years, the study found that the exosomes derived from different sources contained active protein, mRNA, microRNA, long noncoding RNA, and lipid components involved in the biological function of hepatic stellate cells, realized the communication between cells, which played the important regulatory role in the formation of liver fibrosis.ConclusionsExosomes derived from different sources and their contents play an important regulatory role in occurrence and development of liver fibrosis. In the future, exosomes might become a new non-invasive diagnostic method for liver fibrosis to help its early diagnosis, and might also be used as a biological active carrier to achieve its targeted therapy for targeted tissues and cells.