Department of Gastroenterology, The Second Hospital of Hebei Medical University, Hebei Key Laboratory of Gastroenterology, Hebei Institute of Gastroenterology, No. 215 Heping West Road, Shijiazhuang, 050000, Hebei, China.
Dig Dis Sci. 2019 Sep;64(9):2570-2580. doi: 10.1007/s10620-019-05588-5. Epub 2019 Mar 14.
Liver fibrosis is featured with excessive deposition of extracellular matrix and fibrous connective tissue hyperplasia. The specific inhibitor of Raf-1 kinase inhibitor protein, locostatin, inhibits the migration of hepatic stellate cells. In this study, we investigated the effect of locostatin on liver fibrosis and its underlying mechanism.
Carbon tetrachloride (CCl4) was used to induce liver fibrosis in mice, and locostatin was injected intraperitoneally. Liver fibrosis was assessed by Masson and Sirius red staining, hydroxyproline (HYP) assay, and collagen percentage area. Collagen I, collagen III, and α-SMA were detected by RT-PCR and western blot. The levels of MMP-13, MMP-2, TIMP-1, and TIMP-2 were estimated by ELISA. Liver inflammation was evaluated by HE staining and immunohistochemistry; liver myeloperoxidase (MPO), superoxide dismutase, and malondialdehyde were measured by ELISA; and cytokines were by Mouse Cytokine Array Q4000.
Compared to the CCl4 group, HYP (208.56 ± 6.12) µg/g, percentage of total collagen at overall region (1.91 ± 0.13), MMP-13/TIMP-1 (0.19 ± 0.01), MPO (1.45 ± 0.04) U/g, TGF-β (2652 ± 91.20), PDGF-AA (3897 ± 290.69), and E-selectin (1569 ± 66.48) in the liver tissues were decreased significantly in the locostatin-treated group.
Locostatin mitigated liver fibrosis and inflammation induced by CCl4. The mechanism is via inhibition inflammatory cytokines, TGF-β, PDGF-AA, and E-selectin.
肝纤维化的特征是细胞外基质的过度沉积和纤维结缔组织增生。Raf-1 激酶抑制剂蛋白的特异性抑制剂洛司他汀抑制肝星状细胞的迁移。在本研究中，我们研究了洛司他汀对肝纤维化的作用及其机制。
采用四氯化碳（CCl4）诱导小鼠肝纤维化，腹腔注射洛司他汀。通过 Masson 和 Sirius 红染色、羟脯氨酸（HYP）测定和胶原百分比面积评估肝纤维化。通过 RT-PCR 和 Western blot 检测胶原 I、胶原 III 和 α-SMA。通过 ELISA 估计 MMP-13、MMP-2、TIMP-1 和 TIMP-2 的水平。通过 HE 染色和免疫组化评估肝炎症；通过 ELISA 测量肝髓过氧化物酶（MPO）、超氧化物歧化酶和丙二醛；通过 Mouse Cytokine Array Q4000 测定细胞因子。
与 CCl4 组相比，洛司他汀处理组肝组织中的 HYP（208.56±6.12）μg/g、总胶原百分比（1.91±0.13）、MMP-13/TIMP-1（0.19±0.01）、MPO（1.45±0.04）U/g、TGF-β（2652±91.20）、PDGF-AA（3897±290.69）和 E-选择素（1569±66.48）显著降低。
洛司他汀减轻了 CCl4 诱导的肝纤维化和炎症。其机制是通过抑制炎症细胞因子、TGF-β、PDGF-AA 和 E-选择素。
