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Objective To investigate the capillarization of liver sinusoidal endothelial cells (LSECs) and its association with hepatic fibrosis during the development of alveolar echinococcosis, so as to provide the basis for unraveling the mechanisms underlying the role of LSEC in the development and prognosis of hepatic injuries and hepatic fibrosis caused by alveolar echinococcosis. Methods Forty C57BL/6 mice at ages of 6 to 8 weeks were randomly divided into a control group and 1-, 2- and 4-week infection groups, of 10 mice in each group. Each mouse in the infection groups was intraperitoneally injected with 2 000 Echinococcus multilocularis protoscoleces, while each mouse in the control group was given an equal volume of phosphate-buffered saline using the same method. All mice were sacrificed 1, 2 and 4 weeks post-infection and mouse livers were collected. The pathological changes of livers were observed using hematoxylin-eosin (HE) staining, and hepatic fibrosis was evaluated through semi-quantitative analysis of Masson’s trichrome staining-positive areas. The activation of hepatic stellate cells (HSCs) and extracellular matrix (ECM) deposition were examined using immunohistochemical staining of α-smooth muscle actin (α-SMA) and collagen type I alpha 1 (COL1A1), and the fenestrations on the surface of LSECs were observed using scanning electron microscopy. Primary LSECs were isolated from mouse livers, and the mRNA expression of LSEC marker genes Stabilin-1, Stabilin-2, Ehd3, CD209b, GATA4 and Maf was quantified using real-time fluorescence quantitative PCR (qPCR) assay. Results Destruction of local liver lobular structure was observed in mice 2 weeks post-infection with E. multilocularis protoscoleces, and hydatid cysts, which were surrounded by granulomatous tissues, were found in mouse livers 4 weeks post-infection. Semi-quantitative analysis of Masson’s trichrome staining showed a significant difference in the proportion of collagen fiber contents in mouse livers among the four groups (F = 26.060, P < 0.001), and a higher proportion of collagen fiber contents was detected in mouse livers in the 4-week infection group [(11.29 ± 2.58)%] than in the control group (P < 0.001). Immunohistochemical staining revealed activation of a few HSCs and ECM deposition in mouse livers 1 and 2 weeks post-infection, and abundant brown-yellow stained α-SMA and COL1A1 were deposited in the lesion areas in mouse livers 4 weeks post-infection, which spread to surrounding tissues. Semi-quantitative analysis revealed significant differences in α-SMA (F = 7.667, P < 0.05) and COL1A1 expression (F = 6.530, P < 0.05) in mouse levers among the four groups, with higher α-SMA [(7.13 ± 3.68)%] and COL1A1 expression [(13.18 ± 7.20)%] quantified in mouse livers in the 4-week infection group than in the control group (both P values < 0.05). Scanning electron microscopy revealed significant differences in the fenestration frequency (F = 37.730, P < 0.001) and porosity (F = 16.010, P < 0.001) on the surface of mouse LSECs among the four groups, and reduced fenestration frequency and porosity were observed in the 1-[(1.22 ± 0.48)/μm2 and [(3.05 ± 0.91)%] and 2-week infection groups [(3.47 ± 0.10)/μm2 and (7.57 ± 0.23)%] groups than in the control group (all P values < 0.001). There was a significant difference in the average fenestration diameter on the surface of mouse LSECs among the four groups (F = 15.330, P < 0.001), and larger average fenestration diameters were measured in the 1-[(180.80 ± 16.42) nm] and 2-week infection groups [(161.70 ± 3.85) nm] than in the control group (both P values < 0.05). In addition, there were significant differences among the four groups in terms of Stabilin-1 (F = 153.100, P < 0.001), Stabilin-2 (F = 57.010, P < 0.001), Ehd3 (F = 31.700, P < 0.001), CD209b (F = 177.400, P < 0.001), GATA4 (F = 17.740, P < 0.001), and Maf mRNA expression (F = 72.710, P < 0.001), and reduced mRNA expression of Stabilin-1, Stabilin-2, Ehd3, CD209b, GATA4 and Maf genes was quantified in three infection groups than in the control group (all P values < 0.001). Conclusions E. multilocularis infections may induce capillarization of LSECs in mice, and result in a reduction in the expression of functional and phenotypic marker genes of LSECs, and capillarization of LSECs occurs earlier than activation of HSC and development of hepatic fibrosis.
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Liver sinusoidal endothelial cells (LSECs) locate on the surface of hepatic sinusoids. As the first line of defense between the liver and blood, LSECs are the most abundant non-parenchymal cells in the liver. Under physiological conditions, LSECs may induce liver immune tolerance through participating in substance transport and metabolic waste removal, thereby maintaining liver homeostasis, and under pathological conditions, LSECs may promote liver immune response via antigen presentation. LSECs have been found to play a crucial regulatory role in maintaining the balance between liver regeneration and liver fibrosis. This article reviews the progress of researches on LSECs functions, LSECs changes in liver injury, signal pathways associated with regulation of LSECs functions, and the interaction between LSECs and other types of cells in the liver, aiming to elucidate the function of LSECs and their roles in liver diseases.
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Objective:To observe the effect of high glucose downregulated microRNA(miR)-99a on hepatic sinus dysfunction and metformin intervention, and to explore the pathogenesis of diabetes-induced fatty liver and possible mechanism of metformin.Methods:The cultured human liver sinusoidal endothelial cells were randomly divided into normal control group, high glucose model group, miR-99a overexpression group, miR-99a overexpression negative control group, insulin-like growth factor 1 receptor(IGF-1R) inhibitor group, mammalian target of rapamycin(mTOR) inhibitor group, and metformin treatment group. The mRNA expressions of miR-99a were detected with realtime quantitative PCR(RT-qPCR), and the expression levels and distribution of IGF-1R, phosphorylated(p-)mTOR and vitronectin(VN) were detected by Western blotting and immunofluorescence. The ultrastructure of human liver sinusoidal endothelial cells was observed using scanning electron microscope.Results:Compared with normal control group, the mRNA expression of miR-99a was downregulated( P=0.008), while the protein expressions of IGF-1R, mTOR, and VN were significantly increased, and the diameter and number of fenestrae decreased significantly in high glucose model group. Compared with high glucose model group, after the treatment with metformin, the mRNA expression of miR-99a was upregulated, while the protein expressions of IGF-1R, mTOR, and VN were significantly decreased( P=0.001, P=0.016, P=0.005, respectively), the number of fenestras increased and the diameter became larger in miR-99a overexpression group, IGF-1R inhibitor group, mTOR inhibitor group, and metformin treatment group. After overexpression of miR-99a, the protein expressions of IGF-1R, p-mTOR, and VN were significantly reduced( P=0.007, P=0.013, P=0.003, respectively); After administration of IGF-1R inhibitors, the expressions of p-mTOR and VN significantly decreased( P=0.006, P=0.009, respectively), following treatment with the mTOR inhibitor, the expression of VN was significantly reduced( P=0.008), while the expression of IGF-1R remained unchanged( P=0.553). Conclusions:Downregulating of miR-99a with high glucose induced hepatic sinus dysfunction, which may be related to the regulation of IGF-1R/mTOR pathway. Metformin increased the expression of miR-99a, thereby inhibiting high glucose-induced hepatic sinusoidal dysfunction.
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The liver has a unique immune microenvironment, and the intrinsic antigen-presenting cells in the liver interact with each other and form a network to accurately regulate the homeostasis between liver immune tolerance and immune response. During hepatitis B virus (HBV) infection, on the one hand, the intrahepatic intrinsic antigen-presenting cells induce immune tolerance to help the virus escape immune clearance and thus result in persistent infection; on the other hand, the maturation and activation of the intrahepatic intrinsic antigen-presenting cells can also mediate effective anti-HBV immune response to achieve virus clearance. This article elaborates on the research advances in the role and mechanism of action of intrahepatic intrinsic antigen-presenting cells in regulating immune response against HBV infection.
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With the increasing incidence rate of nonalcoholic fatty liver disease (NAFLD) in Western developed countries and rich regions of China year by year, the treatment methods for NAFLD have been constantly improved and NAFLD has become a research hotspot. As an important physiological structure of the liver, liver sinus endothelial cells (LSECs) play an important role in the development and progression of NAFLD. This article summarizes the mechanism of action of LSECs in the pathogenesis of NAFLD, which included the following aspects: LSEC capillarization occurs in nonalcoholic fatty liver and promotes steatosis; meanwhile, LSECs contribute to oxidative stress in nonalcoholic steatohepatitis (NASH) and is a major effector of liver inflammation in NASH, thus promoting liver fibrosis; in addition, angiogenesis is highly stimulated and promotes NAFLD-related hepatocellular carcinoma in NAFLD. However, the role of LSECs in NASH-associated liver cirrhosis has not been validated, which needs to be further clarified to provide new ideas and directions for treatment.
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Objective To evaluate the efficacy of transjugular intrahepatic portosystemic shunt (TIPS)in the treatment of patients with hepatic sinusoidal obstruction syndrome (HSOS).Methods From April 2015 to August 2018,at The First Affiliated Hospital of University of Science and Technology of China,21 patients with gynura segetum caused HSOS were selected.All the patients received TIPS treatment because of unresponsiveness to anticoagulant therapy for at least two weeks.After operation patients were followed up with liver and portal vein Doppler ultrasonography examination,liver and kidney function tests,and survival observation.T test,logistic univariate regression analysis and Cox regression analysis were performed for statistical analysis.Results Among the 21 patients with gynura segetum-related HSOS,18 patients were in the subacute phase and three patients in the chronic phase.All of them were moderate or severe patients and all successfully underwent TIPS.The postoperative portal vein pressure was (16.71 ± 4.68) cmH2O (1 cmH2O =0.098 kPa),which was lower than that before operation ((41.52 ±6.27) cmH2O),and the difference was statistically significant (t =16.936,P ＜ 0.01).The postoperation portal vein blood flow velocity was (41.52 ±7.70) cm/s,which was higher than before operation ((11.19 ± 3.29) cm/s),and the difference was statistically significant (t =-15.191,P ＜0.01).At one month after operation,15 of 21 patients were clinically cured;among the remaining six patients,four patients were improved and two patients were ineffective (including one patient died).At four months after operation,two patients died,and the remaining 19 patients were clinically cured.At one month after operation,the levels of alanine aminotransferase (ALT),aspartate aminotransferase (AST),total bilirubin (TBil) and serum creatinine were (23.7 ± 16.8) U/L,(33.9 ±7.4) U/L,(52.7 ± 38.2) μmol/L and (62.7 ± 12.6) μmol/L,respectively,which were lower than those before operation ((60.5 ± 42.4) U/L,(78.4 ± 42.4) U/L,(74.9 ± 38.2) μmol/L and (82.4 ± 19.6) μmol/L,respectively),and the differences were statistically significant (t =3.193,3.493,2.378 and 4.519;all P ＜ 0.05).The level of albumin was (39.0 ±3.1) g/L,which was higher than that before operation ((30.9 ± 3.8) g/L),and the difference was statistically significant (t =-10.283,P ＜ 0.01).Portal vein thrombosis and preoperative TBil level had predictive value for therapeutic efficacy (both P ＜0.05).The one-year cumulative survival rate of patients was 90.5％.Preoperative TBil level and hepatic encephalopathy had effects on the prognosis of patients (both P ＜ 0.05).Conclusion TIPS is a safe,reliable and effective treatment for patients with subacute and chronic gynura segetum-related HSOS who are not responding to ineffective anticoagulant therapy,which can improve the prognosis and survival rate of the patients.
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<p><b>OBJECTIVE</b>To study the effects of Astragalus polysaccharide (APS), the primary effective component of the Chinese herb medicine Astragalus membranaceus (frequently used for its anti-hepatic fibrosis effects), on nanoscale mechanical properties of liver sinusoidal endothelial cells (SECs).</p><p><b>METHODS</b>Using endothelial cell medium as the control, 5 experimental groups were established utilizing different concentrations of APS, i.e. 12.5, 25, 50, 100, and 200 μg/mL. By using atomic force microscopy along with a microcantilever modified with a silicon dioxide microsphere as powerful tools, the value of Young's modulus in each group was calculated. SAS 9.1 software was applied to analyze the values of Young's modulus at the pressed depth of 300 nm. Environmental scanning electron microscopy was performed to observe the surface microtopography of the SECs.</p><p><b>RESULTS</b>The value of Young's modulus in each APS experimental group was significantly greater than that of the control group: as APS concentration increased, the value of Young's modulus presented as an increasing trend. The difference between the low-concentration (12.5 and 25 μg/mL) and high-concentration (200 μg/mL) groups was statistically significant (P<0.05), but no significant differences were observed between moderateconcentration (50 and 100 μg/mL) groups versus low- or high-concentration groups (P>0.05). Surface topography demonstrated that APS was capable of increasing the total area of fenestrae.</p><p><b>CONCLUSIONS</b>The values of Young's modulus increased along with increasing concentrations of APS, suggesting that the stiffness of SECs increases gradually as a function of APS concentration. The observed changes in SEC mechanical properties may provide a new avenue for mechanistic research of anti-hepatic fibrosis treatments in Chinese medicine.</p>
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Animales , Ratas , Planta del Astrágalo , Química , Fenómenos Biomecánicos , Módulo de Elasticidad , Células Endoteliales , Biología Celular , Hígado , Biología Celular , Microscopía de Fuerza Atómica , Microesferas , Nanotecnología , Polisacáridos , Farmacología , Dióxido de Silicio , Química , Propiedades de SuperficieRESUMEN
To investigate the effects of cryptotanshinone (an active ingredient of Salvia Miltiorrhiza) inhibition of angiogenesis, the toxicity of cryptotanshinone was assayed in human hepatic sinusoidal endothelial cells (HHSEC) by CCK8 method. Max dose without toxicity is 10 μmol·L-1. The proliferation of HHSEC were induced by the endothelial cell growth supplement (ECGS), with 2.5 μmol·L-1 sorafenib as the positive control. Cell proliferation was analyzed by EdU assay. Cell viability was analyzed by CCK8 method. The expression of vWF was analyzed by immunofluorescence method. Fluorescence probe method was used to detect the intracellular nitric oxide (NO) levels. Tube formation of HHSEC and transgenic zebrafish were also observed to evaluate the effects of cryptotanshinone against angiogenesis. Compared with normal control, there is a proliferation of HHSEC induced by ECGS. The expression of vWF and the NO levels increased significantly. Cryptotanshinone inhibited the proliferation, down regulated the expression of vWF and the NO levels. Further, cryptotanshinone inhibited the tube formation of HHSEC and reduced the number of fu nctional vessels in transgenic zebrafish. The results suggest that cryptotanshinone could inhibit angiogenesis by regulating the HHSEC cell function.
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The inducible form of nitric oxide synthase (iNOS) is expressed in hepatic cells in pathological conditions. Its induction is involved in the development of liver fibrosis, and thus iNOS could be a therapeutic target for liver fibrosis. This review summarizes the role of iNOS in liver fibrosis, focusing on 1) iNOS biology, 2) iNOS-expressing liver cells, 3) iNOS-related therapeutic strategies, and 4) future directions.