To investigate the dynamic expression of transforming growth factor-β1 (TGF-β1) and heat shock protein 47 (HSP47) and explore their roles in the progression of hepatic fibrosis induced by Schistosoma japonicum infection.Fifty female mice of the ICR strain were randomly divided into the infection group and the normal control group, of 25 mice in each group. Each mouse in the infection group was infected with 20 ± 1 cercariae of S. japonicum via the abdominal skin, while uninfected animals served as normal control. Five mice were sacrificed 4, 6, 8, 10 and 12 weeks post-infection and liver tissues were sampled. Serum HSP47 and TGF-β1 was determined using enzyme-linked immunosorbent assay (ELISA), and the pathological changes of liver specimens were observed with hematoxylin & eosin (HE) staining. In addition, the synthesis of alpha 1 chain of type I collagen (COL1A1) was measured using Masson staining, and the mRNA expression of TGF-β1, HSP47 and COL1A1 was determined using real-time fluorescent quantitative PCR (qPCR) assay.During the period of S. japonicum-induced hepatic fibrosis, the serum HSP47 and TGF-β1 levels and the mRNA expression of TGF - β1, HSP47 and COL1A1 gradually increased with the progression of hepatic fibrosis. The serum levels of HSP47 and TGF-β1 were (179.26 ± 29.87) pg/mL and (22.37 ± 5.21) ng/mL 6 weeks post-infection, respectively, which were significantly greater than those [(150.29 ± 34.91) pg/mL and (18.54 ± 7.78) ng/mL, respectively] in the normal control group (both P values < 0.05). In addition, the mRNA expression of HSP47, COL1A1 and TGF-β1 was (0.86 ± 0.04), (1.17 ± 0.06) and (0.64 ± 0.13) in mouse liver specimens, which was significantly higher than that (0.23 ± 0.03, 0.20 ± 0.02 and 0.38 ± 0.02) in the normal control group (all P values < 0.01).The expression of TGF-β1 and HSP47 during the period of S. japonicum-induced hepatic fibrosis is consistent with the progression of the hepatic fibrosis, and exhibits the same tendency with type I collagen expression. HSP47 is a novel promising diagnosis marker and therapeutic target for S. japonicum-induced hepatic fibrosis.[摘要] 目的 观察在日本血吸虫诱导的小鼠肝纤维化进程中转化生长因子-β1 (Transforming growth factor-β1, TGF-β1) 和热休克蛋白47 (Heat shock protein 47, HSP47) 的动态表达, 探讨其在日本血吸虫病肝纤维化发生发展中的作用。方 法 将50只雌性ICR小鼠随机分成感染组和正常对照组, 每组25只。感染组每只小鼠经腹部皮肤感染日本血吸虫尾蚴 (20 ± 1) 条, 对照组以不含尾蚴的去氯水处理。分别于感染后4、6、8、10周和12周等5个时间点, 各取5只小鼠肝组织。应用酶联免疫吸附试验检测各小鼠血清中HSP47和TGF-β1含量, 肝组织HE染色观察病理学改变, Masson染色观察胶 原增生情况, 应用实时荧光定量PCR检测肝组织TGF-β1、HSP47、I型胶原 (α1) 链COL1A1 基因mRNA 表达水平。结果 在日本血吸虫诱导小鼠肝纤维化过程中, 小鼠血清HSP47和TGF-β1浓度和肝组织中TGF-β1 mRNA、HSP47 mRNA、COL1A1 mRNA表达水平均随纤维化进展而渐次升高。小鼠感染后6周, 小鼠血清HSP47和TGF-β1含量分别为 (179.26±29.87) pg/mL和 (22.37 ± 5.21) ng/mL, 显著高于同期正常对照组小鼠的 (150.29 ± 34.91) pg/mL和 (18.54 ± 7.78) ng/mL (P 均< 0.05)。肝组织中HSP47 mRNA、COL1A1 mRNA、TGF-β1 mRNA 表达水平分别为 (0.86 ± 0.04)、(1.17 ± 0.06) 和 (0.64 ± 0.13), 均高于同期正常对照组小鼠的 (0.23 ± 0.03)、(0.20 ± 0.02) 和 (0.38 ± 0.02) (P 均< 0.01)。结论 TGF-β1和HSP47 在日本血吸虫诱导的小鼠肝纤维进程中的表达与肝纤维化进程一致, 且随I型胶原的增多呈升高趋势; HSP47有望成为 日本血吸虫病肝纤维化的新诊断标志物和治疗靶点。.
Abstract Background Numerous studies have shown that aberrant microRNA (miRNA) expression is associated with the pathogenesis and progression of various human diseases. Hence, serum miRNAs are considered to be potential biomarkers for the diagnosis of human diseases. This study examined whether several miRNAs known to be commonly deregulated in liver diseases are deregulated in the serum of hosts with hepatic schistosomiasis, and thus whether they could serve as potential markers for detection of schistosome infection and evaluation of the effectiveness of chemotherapy. Methods We analyzed the serum levels of six selected candidate miRNA molecules (miR-146b, miR-122, miR-223, miR-199a-5p, miR-199a-3p, miR-34a) from mice, rabbits, buffalos and humans infected with Schistosoma japonicum using qPCR. We evaluated liver pathology by determining the hydroxyproline content in liver tissues. Primary resident liver cells were isolated to quantify the expression level of deregulated miRNAs. Bioinformatics analyses were also conducted to assess the potential function of miR-223. Results Using a mouse model of Schistosoma japonicum infection, we found that the expression level of serum miR-223 was significantly elevated after infection, but returned to near normal levels after the treatment with praziquantel (PZQ). Importantly, the level of serum miR-223 reflected the extent of liver pathology post-infection. We validated the elevated level of the circulating miR-223 in serum samples of other host species including rabbits, buffalos and humans. In addition, our results showed that miR-223 was primarily located in the Kupffer cells, but its expression levels were significantly up-regulated in hepatocytes, hepatic stellate cells and Kupffer cells after infection. Bioinformatics analyses revealed a potential functional role of miR-223 in transcription regulator activity, transcription factor activity and DNA binding. Conclusions This study suggested that the circulating miR-223 could serve as a potential new biomarker for the detection of schistosome infection and the assessment of the response to chemotherapy.
Schistosomiasis is a serious parasitic disease in humans, which can lead to liver fibrosis and death. Accumulating evidence indicated that targeting the deregulated microRNAs (miRNAs) could mitigate disease outcomes. Here, we showed that progressive hepatic schistosomiasis caused elevation of miR‐21 and efficient and sustained inhibition of miR‐21 by using highly hepatic tropic adeno‐associated virus serotype 8 (rAAV8), which protected mice against lethal schistosome infection through attenuation of hepatic fibrosis (HF). We demonstrated an additive role of interleukin (IL)−13 and transforming growth factor beta 1 (TGF‐β1) in up‐regulating miR‐21 expression in hepatic stellate cells (HSCs) by activation of mothers against decapentaplegic (SMAD) proteins. Furthermore, down‐regulation of miR‐21 in HSCs reversed HF by enhancing SMAD7 expression, thus repressing TGF‐β1/Smad and IL‐13/Smad pathways. Conclusion : This study suggests the mechanism of IL‐13‐mediated schistosomiasis HF by up‐regulation of miR‐21 and highlights the potential of rAAV8‐mediated miR‐21 inhibition as a therapeutic intervention for hepatic fibrotic diseases, such as schistosomiasis. (H epatology 2015;61:2008–2017)
Liver fibrosis is a common outcome of chronic liver injuries including Schistosoma infection.Activation of resident hepatic stellate cells (HSC) into proliferative,contractile,and fibrogenic myofibrosblast is considered the key process driving liver fibrosis.Recent studies have highlighted that HSC plays an important role in both murine and human schistosomiasis liver fibrosis.Studies of the HSC and its interaction with Schistosoma eggs may be pivotal in our understanding of the pathology associated with schistosomiasis liver fibrosis and provide new approaches to prevent and treat schistosomiasis liver fibrosis.
Key words:
Hepatic stellate cells; Liver fibrosis; Schistosomiasis
Liver fibrosis in schistosomiasis is a serious pathological consequence from immune reactions to schistosome infection. The progression of liver fibrosis depends on the state of immune response. Recent studies have found that Th17 and Treg cells are two subsets of CD4+T cells. The Th17 cells are mainly involved in inflammatory responses, while the Treg cells mainly mediate downregulation of the responses. Under normal conditions, the differentiations of the two subsets are inhibited by each other, and they function oppositely. The balance between Th17 and Treg cells, as well as the balance between them, play an important role in the maintenance of homeostasis and are involved in inflammatory responses, tissue trauma, fibrosis and development of many diseases. This paper reviews the role of Th17/Treg cells and their imbalance in liver fibrosis in schistosomiasis.
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