New gene therapy strategies for hepatic fibrosis

The liver is the largest internal organ of the body, which may suffer acute or chronic injury induced by many factors, leading to cirrhosis and hepatocarcinoma. Cirrhosis is the irreversible end result of fibrous scarring and hepatocellular regeneration, characterized by diffuse disorganization of the normal hepatic structure, regenerative nodules and fibrotic tissue. Cirrhosis is associated with a high co-morbidity and mortality without effective treatment, and much research has been aimed at developing new therapeutic strategies to guarantee recovery. Liver-based gene therapy has been used to downregulate specific genes, to block the expression of deleterious genes, to delivery therapeutic genes, to prevent allograft rejection and to augment liver regeneration. Viral and non-viral vectors have been used, with viral vectors proving to be more efficient. This review provides an overview of the main strategies used in liver-gene therapy represented by non-viral vectors, viral vectors, novel administration methods like hydrodynamic injection, hybrids of two viral vectors and blocking molecules, with the hope of translating findings from the laboratory to the patient′s bed-side.

Molecular therapy for hepatic injury and fibrosis:Where are we?

Hepatic fibrosis is a wound healing response, involving pathways of inflammation and fibrogenesis. In response to various insults, such as alcohol, ischemia, viral agents, and medications or hepatotoxins, hepatocyte damage will cause the release of cytokines and other soluble factors by Kupffer cells and other cell types in the liver. These factors lead to activation of hepatic stellate cells, which synthesize large amounts of extracellular matrix components. With chronic injury and fibrosis, liver architecture and metabolism are disrupted, eventually manifesting as cirrhosis and its complications. In addition to eliminating etiology, such as antiviral therapy and pharmacological intervention, it is encouraging that novel strategies are being developed to directly address hepatic injury and fibrosis at the subcellular and molecular levels. With improvement in understanding these mechanisms and pathways, key steps in injury, signaling, activation, and gene expression are being targeted by molecular modalities and other molecular or gene therapy approaches. This article intends to provide an update in terms of the current status of molecular therapy for hepatic injury and fibrosis and how far we are from clinical utilization of these new therapeutic modalities.

Oxymatrine liposome attenuates hepatic fibrosis via targeting hepatic stellate cells

AIM： To investigate the potential mechanism of Arg- Gly-Asp （RGD） peptide-labeled liposome loading oxy- matrine （OM） therapy in CCI4-induced hepatic fibrosis in rats. METHODS： We constructed a rat model of CCh- induced hepatic fibrosis and treated the rats with dif- ferent formulations of OM. To evaluate the antifibrotic effect of OM, we detected levels of alkaline phospha- tase, hepatic histopathology （hematoxylin and eosin stain and Masson staining） and fibrosis-related gene expression of matrix metallopeptidase （MMP）-2, tis- sue inhibitor of metalloproteinase （TIMP）-I as well as type I procollagen via quantitative real-time poly- merase chain reaction. To detect cell viability and apop- tosis of hepatic stellate cells （HSCs）, we performed 3-（4,5）-dimethylthiahiazo（-z-yl）-3,5-diphenytetrazoli- umromide assay and flow cytometry. To reinforce the combination of oxymatrine with HSCs, we constructed fluorescein-isothiocyanate-conjugated Arg-Gly-Asp peptide-labeled liposomes loading OM, and its targeting of HSCs was examined by fluorescent microscopy. RESULTS： OM attenuated CCh-induced hepatic fibro- sis, as defined by reducing serum alkaline phosphatase （344.47± 27.52 U/L vs 550.69 ± 43.78 U/L, P 〈 0.05）, attenuating liver injury and improving collagen deposits （2.36% ± 0.09% vs 7.70% ±0.60%, P 〈 0.05） and downregulating fibrosis-related gene expression, that is, MMP-2, TIMP-1 and type I procollagen （P 〈 0.05）. OM inhibited cell viability and induced apoptosis of HSCs in vitro. RGD promoted OM targeting of HSCs and en- hanced the therapeutic effect of OM in terms of serum alkaline phosphatase （272.51 ± 19.55 U/L vs 344.47 ± 27.52 U/L, P 〈 0.05）, liver injury, collagen deposits （0.26%± 0.09% vs 2.36% ± 0.09%, P 〈 0.05） and downregulating fibrosis-related gene expression, that is, MMP-2, TIMP-1 and type I procollagen （P 〈 0.05）. Moreover, in vitro assay demonstrated that RGD en- hanced the effect of OM on HSC viability and apoptosis. CONCLUSION： OM attenuated hepatic fibrosis by in- hibiting viability and inducing apoptosis of HSCs. The RGD-labeled formulation enhanced the targeting effi- ciency for HSCs and the therapeutic effect.

Effects of augmentation of liver regeneration recombinant plasmid on rat hepatic fibrosis

AIM: To investigate the effects of eukaryotic expression of plasmid on augmentation of liver regeneration (ALR) in rat hepatic fibrosis and to explore their mechanisms. METHODS: Ten rats were randomly selected from 50 Wistar rats as normal control group. The rest were administered intraperitoneally with porcine serum twice weekly. After 8 wk, they were randomly divided into: model control group, colchicine group (Col), first ALR group (ALR1), second ALR group (ALR2). Then colchicine ALR recombinant plasmid were used to treat them respectively. At the end of the 4th wk, rats were killed. Serum indicators were detected and histopathological changes were graded. Expression of type Ⅰ, Ⅲ, collagen and TIMP-1 were detected by immunohisto-chemistry and expression of TIMP-1 mRNA was detected by semi-quantified RT-PCR. RESULTS: The histologic examination showed that the degree of the rat hepatic fibrosis in two ALR groups was lower than those in model control group. Compared with model group, ALR significantly reduced the serum levels of ALT, AST, HA, LN, PCIII and IV (P<0.05). Immunohistochemical staining showed that expression of type Ⅰ, Ⅲ, collagen and TIMP-1 in two ALR groups was ameliorated dramatically compared with model group (I collagen: 6.94±1.42,5.80±1.66 and 10.83±3.58 in ALR1, ALR2 and model groups, respectively; Ⅲ collagen: 7.18±1.95, 4.50±1.67 and 10.25±2.61, respectively; TIMP-1: 0.39±0.05,0.20±0.06 and 0.53±0.12, respectively,P<0.05 or P<0.01). The expression level of TIMP-1 mRNA in the liver tissues was markedly decreased in two ALR groups compared with model group (TIMP-1 mRNA/β-actin: 0.89±0.08, 0.65±0.11 and 1.36±0.11 in ALR1, ALR2 and model groups respectively, P<0.01). CONCLUSION: ALR recombinant plasmid has beneficial effects on rat hepatic fibrosis by enhancing regeneration of injured liver cells and inhibiting TIMP-1 expressions.

Assessing significant fibrosis using imaging-based elastography in chronic hepatitis B patients: Pilot study

BACKGROUND Accurate detection of significant fibrosis(fibrosis stage 2 or higher on the METAVIR scale)is important especially for chronic hepatitis B(CHB)patients with high viral loads but with normal or mildly elevated alanine aminotransferase(ALT)levels because the presence of significant fibrosis is accepted as the indication for antiviral treatment.Liver biopsy is the reference standard for diagnosing significant fibrosis,but it is an invasive procedure.Consequently,noninvasive imaging-based measurements,such as magnetic resonance elastography(MRE)or two-dimensional shear-wave elastography(2DSWE),have been proposed for the quantitative assessment of liver fibrosis.AIM To explore MRE and 2D-SWE to identify fibrosis stage,and to compare their performance with that of serum-based indices.METHODS The study enrolled 63 treatment-na?ve CHB patients with high viral loads but with normal or mildly elevated ALT levels who underwent liver biopsy before a decision was made to initiate antiviral therapy.MRE and 2D-SWE were performed,and serum-based indices,such as FIB-4 and aspartate transaminase to platelet ratio index(APRI),were calculated.The diagnostic performances of MRE,2D-SWE,FIB-4,and APRI for assessing significant fibrosis(≥F2)and cirrhosis(F4)were evaluated with liver histology as the reference standard,using receiver operating characteristic analyses.RESULTS The liver fibrosis stage was F0/F1 in 19,F2 in 14,F3 in 14,and F4 in 16 patients,respectively.MRE significantly discriminated F2 from F0/1(P=0.022),whereas 2D-SWE showed a broad overlap in distinguishing those stages.MRE showed a higher correlation coefficient value with fibrosis stage than 2D-SWE with fibrosis stage(0.869 vs 0.649,Spearman test;P<0.001).Multivariate linear regression analyses showed that fibrosis stage was the only factor affecting the values of MRE(P<0.001),whereas body mass index(P=0.042)and fibrosis stage(P<0.001)were independent factors affecting 2D-SWE values.MRE performance for diagnosing significant fibrosis was better[area under the curve(AUC)=0.906,positive predictive value(PPV)97.3%,negative predictive value(NPV)69.2%]than that of FIB-4(AUC=0.697,P=0.002)and APRI(AUC=0.717,P=0.010),whereas the performance of 2D-SWE(AUC=0.843,PPV 86%,NPV 65%)was not significantly different from that of FIB-4 or APRI.CONCLUSION Compared to SWE,MRE might be more precise non-invasive assessment for depicting significant fibrosis and for making-decision to initiate antiviral-therapy in treatment-na?ve CHB patients with normal or mildly-elevated ALT levels.

Encapsulating taurine into liposomes:A promising therapeutic for liver fibrosis

We summarize the mechanism by which taurine(Tau)inhibits autophagy and induces iron apoptosis in hepatic stellate cells.Tau interacts with autophagy regulates multifunctional proteins,microtubule-associated protein 1 light chain 3 Beta,and autophagy-related gene 5 to inhibit autophagy,binds to ferritin heavy chain 1 and nuclear receptor coactivator 4 to trigger ferritin autophagy,and interacts with glutathione peroxidase 4 to promote iron apoptosis.There is a solid rationale for developing Tau-based therapies targeting autophagy and ferroptosis regulation.From a pharmaceutical point of view,there are certain requirements for Tau protein delivery systems,such as loading efficiency,stability,and targeting.Nanomaterials should also contain a hydrophilic motif similar to Tau to optimize loading efficiency.Since Tau is a hydrophilic molecule with high water solubility,liposomes,micelles,and amphiphilic polymer nanoparticles may represent a superior choice.The nanostructure of the liposome includes a water region and a lipid membrane to sequester hydrophilic and hydrophobic drugs,respectively,whereas Tau is expected to be loaded into the water region.In addition,a representative method of actively targeting hematopoietic stem cells is introduced.A Tau-based method for the treatment of liver fibrosis is proposed based on the formulation of common liposomes(lecithin plus cholesterol).

Mechanism of action and targeted therapy of stellate cells in liver fibrosis

The incidence of liver fibrosis is increasing worldwide,and if left untreated,it will later develop into cirrhosis with a high mortality rate.In this paper,the activation pathway and related mechanism of stellate cells in liver fibrosis are introduced,and some current therapeutic methods are summarized.These results suggest that stellate cells play an important role in liver fibrosis,and targeted therapy for the purpose of inhibiting the activation of stellate cells and inducing their apoptosis is expected to be an effective regimen to reverse liver fibrosis.However,there are some problems such as insufficient in-depth study of related mechanisms and imperfect experiments.In future animal experiments and clinical trials,more studies can be carried out to provide high-quality protocols for the treatment of liver fibrosis.

Effects and mechanism of adenovirus-mediated phosphatase and tension homologue deleted on chromosome ten gene on collagen deposition in rat liver fibrosis

AIM To evaluate the effects of phosphatase and tension homologue deleted on chromosome ten(PTEN) gene on collagen metabolism in hepatic fibrosis and the underlying mechanisms.METHODS rat primary hepatic stellate cells(HSCs) and human LX-2 cells were transfected with adenovirus containing c DNA constructs encoding wild-type PTEN(Ad-PTEN), PTEN mutant G129 E gene(Ad-G129E), and r NA interference constructs targeting the PTEN sequence PTEN short hairpin r NA to up-regulate and downregulate the expression of PTEN. HSCs were assayed using fluorescent microscopy, real-time polymerase chain reaction, and western blotting. Moreover, a CCl_4-induced rat hepatic fibrosis model was established to investigate the in vivo effects. Hematoxylin and eosin, and Masson's trichrome were used to assess the histological changes. The expression of collagen Ⅰ and Ⅲ was assessed using immunohistochemistry and western blot analysis.RESULTS Elevated expression of PTEN gene reduced serum levels of alanine transaminase and aspartate transaminase, decreased collagen deposition in the liver, and reduced hepatocyte necrosis. In contrast, knockdown of PTEN expression had an opposite effect, such as increased collagen deposition in the liver, and was molecularly characterized by the increased expression of matrix metalloproteinase(MMP)-13(P < 0.01) and MMP-2(P < 0.01), as well as decreased expression of the tissue inhibitor of metalloproteinase(TIMP)-1(P < 0.01) and TIMP-2(P < 0.01).CONCLUSION These data indicated that gene therapy using recombinant adenovirus encoding PTEN might be a novel way of treating hepatic fibrosis.

Inhibiting effect of antisense oligonucleotides phosphorthioate on gene expression of TIMP-1 in rat liver fibrosis

AIM: To observe the inhibition of antisense oligonucleotides (asON) phosphorthioate to the tissue inhibitors metalloproteinase-1 (TIMP-1) gene and protein expression in the liver tissue of immunologically induced hepatic fibrosis rats. The possibility of reversing hepatic fibrosis through gene therapy was observed. METHODS: Human serum albumin (HSA) was used to attack rats, as hepatic fibrosis model, in which asONs were used to block the gene and protein expressing TIMP-1. According to the analysis of modulator, structure protein, coding series of TIMP-1 genome, we designed four different asONs. These asONs were injected into the hepatic fibrosis models through coccygeal vein. The results was observed by RT-PCR for measuring TIMP-1 mRNA expression, immunohistochemistry and in situ hybridization for collagen I, II, special staining of collagen fiber, and electron microscopic examination. RESULTS: Hepatic fibrosis could last within 363 days in our modified model. The expressing level of TIMP-1 was high during hepatic fibrosis process. It has been proved by the immunohistochemical and the electron microscopic examination that the asON phosphorthioate of TIMP-1 could exactly express in vivo. The effect of colchicine was demonstrated to inhibit the expressing level of mRNA and the content of collagen I, III in the liver of experimental hepatic fibrosis rats. However, the electron microscopy research and the pathologic grading of hepatic fibrosis showed that there was no significant difference between the treatment group and the model group (P】 0.05). CONCLUSION: The experimental rat model of hepatic fibrosis is one of the preferable models to estimate the curative effect of anti-hepatic fibrosis drugs. The asON phosphorthioate of TIMP-1 could block the gene and protein expression of TIMP-1 in the liver of experimental hepatic fibrosis rats at the mRNA level. It is possible to reverse hepatic fibrosis, and it is expected to study a new drug of antihepatic fibrosis on the genetic level. Colchicine has very limited therapeutic effect on hepatic fibrosis, furthermore, its toxicity and side effects are obvious.

Evaluation of the prognostic value of liver stiffness in patients with hepatitis C virus treated with triple or dual antiviral therapy:A prospective pilot study

AIM:To evaluate the association between liver stiffness(LS) prior to the initiation of dual/triple therapy and viral response.METHODS:LS was measured in all patients before treatment was administered.The therapeutic approach was based on hepatic,virological,and immunological evaluations and considered the fact that patients with severe fibrosis(F3)or compensated cirrhosis(F4)in Child-Pugh class A are the primary candidates for triple therapy.In total,65 hepatitis C virus(HCV)patients were treated with Peg-interferon/ribavirin(Peg-IFN/RBV);24patients were classified as genotypes 1/4(36.92%),and41 patients were classified as genotypes 2/3(63.08%)(dual therapy).In addition,20 HCV treatment-experienced genotype 1 patients were treated with Peg IFN-RBV and boceprevir(triple therapy).Wilcoxon rank-sum tests were used to compare the groups.RESULTS:LS significantly differed between dual therapy and triple therapy(P=0.002).The mean LS value before dual therapy treatment was 8.61±5.79k Pa and was significantly different between patients achieving a sustained virologic response(SVR)24weeks after therapy and those who did not(7.23±5.18 k Pa vs 11.72±5.99 k Pa,respectively,P=0.0003).The relative risk of non-response to therapy was 4.45(95%CI:2.32-8.55).The attributable risk of non-response to therapy was 49%.The mean LSvalue before triple therapy treatment was 13.29±8.57k Pa and was significantly different between patients achieving and not achieving SVR24(9.41±5.05 vs19.11±9.74,respectively;P=0.008).The relative risk of non-response to therapy was 5.57%(95%CI:1.50-20.65).The attributable risk of non-response to therapy(70%)was increased compared with dual therapy patients.Pre-treatment stiffness>12 k Pa was significantly associated with non-SVR(P<0.025)in both groups.CONCLUSION:Pre-treatment liver stiffness may be useful for predicting the response to treatment in patients treated with either dual or triple anti-HCV therapy.

Guidelines for diagnosis and treatment of hepatic fibrosis with integrated traditional Chinese and Western medicine(2019 edition)

In 2006, the Hepatology Committee of Chinese Association of Integrative Medicine issued the "Guidelines for the Prevention and Treatment of Liver Fibrosis with Integrated Traditional Chinese and Western Medicine." In recent years, the fields of Chinese medicine, Western medicine, and integrative medicine have made rapid advances in basic and clinical research into chronic liver disease, and accumulated new evidence for the prevention and treatment of hepatic fibrosis. Therefore, in order to meet clinical needs, liver disease experts of integrated traditional Chinese and Western medicine were united to revise the previous guidelines in order to help physicians make correct and reasonable decisions in the diagnosis and treatment of hepatic fibrosis.

Sustained virological response: A milestone in the treatment of chronic hepatitis C

AIM: To evaluate the long-term eradication of hepatitis C virus (HCV) infection and liver-related complications in chronically infected patients that have achieved sustained virological response. METHODS: One hundred and fifty subjects with chronic hepatitis C (CHC) or cirrhosis and sustained virological response (SVR) between the years of 1989 and 2008 were enrolled in a long-term clinical follow-up study at the Gastrointestinal and Liver Unit of the University Hospital of Naples "Federico Ⅱ". At the beginning of the study, the diagnosis of HCV infection was made on the basis of serum positivity for antibodies to HCV and detection of HCV RNA transcripts, while a diagnosis of chronic hepatitis was formulated using imaging techniques and/or a liver biopsy. SVR was achieved by interferon-based therapy, both conventional and pegylated, with and without ribavirin treatment. The patients were evaluated for follow-up at a median length of 8.6 years, but ranged from 2-19.9 years. Among them, 137 patients had pre-treatment CHC and 13 had cirrhosis. The patients were followed with clinical, biochemical, virological, and ultrasound assessments on a given schedule. Finally, a group of 27 patients underwent a liver biopsy at the beginning of the study and transient elastography at their final visit to evaluate changes in liver fibrosis. RESULTS: The median follow-up was 8.6 years (range 2-19.9 years). HCV RNA remained undetectable in all patients, even in patients who eventually developed liver-related complications, indicating no risk of HCV recurrence. Three liver-related complications were observed: two cases of hepatocellular carcinoma and one case of bleeding from esophageal varices resulting in an incidence rate of 0.23%/person per year. Further, all three complications took place in patients diagnosed with cirrhosis before treatment began. Only one death due to liver-related causes occurred, resulting in a mortality rate of 0.077% person per year. This amounts to a 99.33% survival rate in our cohort of patients after therapy for HCV infection. Finally, of the 27 patients who underwent a liver biopsy at the beginning of the study, a reduction in liver fibrosis was observed in 70.3% of the cases; only three cases registering values of liver stiffness indicative of significant fibrosis. CONCLUSION: Patients with CHC and SVR show an excellent prognosis with no risk of recurrence and a very low rate of mortality. Our data indicate that viruseradication following interferon treatment can last up to 20 years.

A small molecule fibrokinase inhibitor in a model of fibropolycystic hepatorenal disease

AIM To evaluate the novel platelet-derived growth factor receptor and vascular endothelial growth factor receptor dual kinase inhibitor ANG3070 in a polycystic kidney disease-congenital hepatic fibrosis model.METHODS At 6 wk of age, PCK rats were randomized to vehicle or ANG3070 for 4 wk. At 10 wk, 24 h urine and left kidneys were collected and rats were continued on treatment for 4 wk. At 14 wk, 24 h urine was collected, rats were sacrificed, and liver and right kidneys were collected for histological evaluation. For Western blot studies, PCK rats were treated with vehicle or ANG3070 for 7 d and sacrificed approximately 30 min after the last treatments.RESULTS Compared to the wild-type cohort, the PCK kidney(Vehicle cohort) exhibited a marked increase in kidney and liver mass, hepato-renal cystic volume, hepato-renal fibrosis and hepato-renal injury biomarkers. Intervention with ANG3070 in PCK rats decreased kidney weight, reduced renal cystic volume and reduced total kidney hydroxyproline, indicating significantly reduced rental interstitial fibrosis compared to the PCK-Vehicle cohort. ANG3070 treatment also mitigated several markers of kidney injury, including urinary neutrophil gelatinase-associated lipocalin, kidney injury molecule-1, cystatin C and interleukin-18 levels. In addition, this treatment attenuated key indices of renal dysfunction, including proteinuria, albuminuria and serum blood urea nitrogen and creatinine, and significantly improved renal function compared to the PCK-Vehicle cohort. ANG3070 treatment also significantly decreased liver enlargement, hepatic lesions, and liver fibrosis, and mitigated liver dysfunction compared to the PCK-Vehicle cohort.CONCLUSION These results suggest that ANG3070 has the potential to slow disease, and may serve as a bridge toward hepato-renal transplantation in patients with fibropolycystic disease.

Current treatment for hepatitis C virus/human immunodeficiency virus coinfection in adults

Hepatitis C virus(HCV)/human immunodeficiency virus(HIV)coinfection is a major problem among HIV-infected patients,resulting in increased morbidity and mortality rates due to the acceleration of liver fibrosis progression by HIV,leading to liver cirrhosis and hepatocellular carcinoma.Although the efficacy of directacting antiviral therapy in patients with HIV/HCV coinfection and HCV monoinfection are similar in terms of sustained virologic response rate,there are some additional complications that arise in the treatment of patients with HIV/HCV coinfection,including drug-drug interactions and HCV reinfection due to the high risk behavior of these patients.This review will summarize the current management of HIV/HCV coinfection.

Nationwide retrospective study of hepatitis B virological response and liver stiffness improvement in 465 patients on nucleos(t)ide analogue

BACKGROUND Hepatitis B virus(HBV)nucleos(t)ide analog(NA)therapy reduces liver disease but requires prolonged therapy to achieve hepatitis B surface antigen(HBsAg)loss.There is limited North American real-world data using non-invasive tools for fibrosis assessment and few have compared 1st generation NA or lamivudine(LAM)to tenofovir disoproxil fumarate(TDF).AIM To assess impact of NA on virological response and fibrosis regression using liver stiffness measurement(LSM)(i.e.,FibroScan®).METHODS Retrospective,observational cohort study from the Canadian HBV Network.Data collected included demographics,NA,HBV DNA,alanine aminotransferase(ALT),and LSM.Patients were HBV monoinfected patients,treatment naïve,and received 1 NA with minimum 1 year follow-up.RESULTS In 465(median 49 years,37%female,35%hepatitis B e antigen+at baseline,84%Asian,6%White,and 9%Black).Percentage of 64(n=299)received TDF and 166 were LAM-treated with similar median duration of 3.9 and 3.7 years,respectively.The mean baseline LSM was 11.2 kPa(TDF)vs 8.3 kPa(LAM)(P=0.003).At 5-year follow-up,the mean LSM was 7.0 kPa in TDF vs 6.7 kPa in LAM(P=0.83).There was a significant difference in fibrosis regression between groups(i.e.,mean-4.2 kPa change in TDF and-1.6 kPa in LAM,P<0.05).The last available data on treatment showed that all had normal ALT,but more TDF patients were virologically suppressed(<10 IU/mL)(n=170/190,89%)vs LAM-treated(n=35/58,60%)(P<0.05).None cleared HBsAg.CONCLUSION In this real-world North American study,approximately 5 years of NA achieves liver fibrosis regression rarely leads to HBsAg loss.

丙酚替诺福韦联合肝爽颗粒治疗慢性乙型肝炎患者疗效及其对肝纤维化指标的影响

目的观察丙酚替诺福韦(TAF)联合肝爽颗粒治疗慢性乙型肝炎(CHB)患者的疗效,并重点评估肝纤维化指标的变化。方法2020年1月~2023年12月我院诊治的76例CHB患者被随机分为对照组38例和观察组38例,分别给予TAF治疗或TAF联合肝爽颗粒治疗,在治疗48周末评估疗效。采用实时荧光定量PCR法检测血清HBV DNA载量,采用化学发光免疫分析法检测血清HBeAg水平,计算肝纤维化4因子指数(FIB-4),使用瞬时弹性成像仪行肝硬度检测(LSM),采用ELISA法检测血清转化生长因子β1(TGF-β1)、基质金属蛋白酶1(MMP-1)、金属蛋白酶组织抑制因子1(TIMP-1)水平。结果在治疗48 w末,观察组血清ALT复常率为94.7%,显著高于对照组的78.9%(P<0.05);观察组胁肋胀痛、舌苔黄腻、纳呆呕恶和尿黄等中医评分分别为(0.9±0.1)分、(1.0±0.2)分、(0.9±0.1)分和(0.7±0.1)分,均显著低于对照组【分别为(1.3±0.2)分、(1.3±0.2)分、(1.2±0.2)分和(0.9±0.2)分,均P<0.05】;观察组FIB-4、LSM、血清TGF-β1和TIMP-1水平分别为(1.8±0.3)、(7.0±0.6)kPa、(21.4±5.4)ng/mL和(119.3±19.5)ng/mL,均显著低于对照组【分别为(2.2±0.4)、(9.7±1.1)kPa、(39.1±6.1)ng/mL和(168.9±22.3)ng/mL,P<0.05】,而血清MMP-1水平为(9.8±1.2)ng/mL,显著高于对照组【(7.1±1.0)ng/mL,P<0.05】。结论应用丙酚替诺福韦联合肝爽颗粒治疗CHB患者可显著减轻中医证候评分,改善肝功能,减轻肝纤维化,值得进一步验证。

血清骨桥蛋白,血管生成素样蛋白8水平与原发性肝癌病人介入治疗后肝纤维化的相关性

目的探讨原发性肝癌(PHC)病人血清骨桥蛋白(OPN)、血管生成素样蛋白8(ANGPTL8)水平与经导管动脉化疗栓塞术(TACE)后肝纤维化(HF)的相关性。方法2021年3月~2023年6月期间收治的PHC病人166例,根据介入治疗后是否发生HF分为两组,其中发生HF的病人92例,为观察组,未发生HF的病人74例,为对照组;酶联免疫吸附法(ELISA)测定血清OPN、ANGPTL8水平并分析OPN、ANGPTL8水平对HF的预测价值。采用Pearson相关性分析OPN、ANGPTL8水平与生化指标的相关性;采用多因素Logistics回归分析影响HF发生的因素;采用ROC曲线分析OPN、ANGPTL8对HF的预测价值。结果观察组血清OPN[(74.56±11.56)ng/ml]、ANGPTL[(42.78±5.23)ng/ml]、ALT[(62.24±9.56)U/L]、AST[(42.88±8.23)U/L]、HA[(252.98±52.44)ng/L]、LN[(152.64±26.45)ng/L]、PCⅢ[(16.54±3.46)ng/L]、Ⅳ-C[(152.78±21.23)ng/L]水平明显高于对照组[(57.89±9.68)ng/ml、(35.46±4.78)ng/ml、(49.46±7.46)U/L、(31.48±7.26)U/L、(192.56±23.88)ng/L、(124.48±11.23)ng/L、(11.26±2.23)ng/L、(126.45±18.56)ng/L],两组比较,差异有统计学意义(P<0.05)。血清OPN、ANGPTL8及二者联合预测HF发生的AUC分别为0.914、0.920、0.978,OPN联合ANGPTL8预测HF发生的AUC高于二者分别单独预测(P<0.05)。结论PHC病人血清OPN、ANGPTL8水平与HF的发生密切相关,二者是HF发生的影响因素,可作为预测HF发生的指标。

Interplays of liver fibrosis-associated microRNAs:Molecular mechanisms and implications in diagnosis and therapy

microRNAs(miRNAs)are a class of non-coding functional small RNA composed of 21e23 nucleotides,having multiple associations with liver fibrosis.Fibrosis-associated miRNAs are roughly classified into pro-fibrosis or anti-fibrosis types.The former is capable of activating hepatic stellate cells(HSCs)by modulating pro-fibrotic signaling pathways,mainly including TGF-b/SMAD,WNT/b-catenin,and Hedgehog;the latter is responsible for maintenance of the quiescent phenotype of normal HSCs,phenotypic reversion of activated HSCs(aHSCs),inhibition of HSCs proliferation and suppression of the extracellular matrix-associated gene expression.Moreover,several miRNAs are involved in regulation of liver fibrosis via alternative mechanisms,such as interacting between hepatocytes and other liver cells via exosomes and increasing autophagy of aHSCs.Thus,understanding the role of these miRNAs may provide new avenues for the development of novel interventions against hepatic fibrosis.

Elastography for Longitudinal Assessment of Liver Fibrosis after Antiviral Therapy:A Review

Chronic hepatitis B or C viral infection is a common cause of liver cirrhosis and hepatocellular carcinoma. Fibrosis regres-sion can be achieved after long-term antiviral therapy (AVT). Monitoring of dynamic changes in liver fibrosis after treat-ment is essential for establishing prognosis and formulation of a follow-up surveillance program. Routine surveillance of fibrosis after AVT by liver biopsy, the gold standard for fibrosis assessment, is hindered by its invasive nature, sampling error and observer variability. Elastography is a noninvasive quan-titative alternative that has been widely used and validated for the staging of liver fibrosis prior to treatment. Recently, increasing research interest has been focused on the role of elastography in longitudinal assessment of liver fibrosis after AVT. In this review, the basic principles, acquisition techni-ques, diagnostic performances, and strengths and limitations of ultrasound elastography and magnetic resonance elastog-raphy are presented. Emerging evidence regarding the use of elastography techniques for the monitoring of liver fibrosis after AVT is summarized. Current challenges and future directions are also discussed, designed to optimize the application of these techniques in clinical practice.

水飞蓟素联合复方蛋氨酸胆碱治疗酒精性肝炎患者疗效及其对血清肝纤维化指标的影响

目的观察应用水飞蓟素联合复方蛋氨酸胆碱治疗酒精性肝炎(AH)患者的临床疗效及其对血清肝纤维化指标的影响。方法2021年1月~2023年1月我院诊治的AH患者101例,采用随机数字表法将患者分为对照组50例和观察组51例,分别给予复方蛋氨酸胆碱或复方蛋氨酸胆碱联合水飞蓟素治疗6个月。采用放射免疫分析法检测血清层粘连蛋白(LN)、透明质酸(HA)、Ⅲ型前胶原N端肽(PCⅢ)和Ⅳ型胶原(Ⅳ-C),采用ELISA法检测血清过氧化物酶体增殖物激活受体γ(PPARγ)、瘦素(LEP)、脂联素(APN)和白细胞介素-18(IL-18)水平。结果经治疗,观察组血清ALT和AST水平分别为(41.8±4.9)IU/L和(42.5±4.9)IU/L,均显著低于对照组【分别为(75.3±6.8)IU/L和(62.7±5.8)IU/L,P<0.05】;观察组血清HA、PCⅢ和Ⅳ-C水平分别为(103.6±13.5)μg/L、(94.2±9.8)μg/L和(75.2±8.6)μg/L,均显著低于对照组【分别为(146.2±15.3)μg/L、(125.8±13.3)μg/L和(112.3±14.5)μg/L,P<0.05】;观察组血清PPARγ、LEP和IL-18水平分别为(231.7±26.8)pg/mL、(5.4±0.7)μg/L和(92.5±11.7)ng/mL,均显著低于对照组【分别为(285.3±29.4)pg/mL、(8.3±0.9)μg/L和(118.5±12.4)ng/mL,P<0.05】,而血清APN水平为(15.4±1.7)ng/L,显著高于对照组【(11.3±1.9)ng/L,P<0.05】。结论应用水飞蓟素联合复方蛋氨酸胆碱治疗AH患者效果较好,有利于肝功能指标恢复,可能对延缓肝纤维化进展、降低机体炎症反应和调节脂代谢有帮助。