Hirudin Protects Ang II-Induced Myocardial Fibroblasts Fibrosis by Inhibiting the Extracellular Signal-Regulated Kinase1/2 (ERK1/2) Pathway

Abstract

BackgroundMyocardial fibrosis is closely related to all types of cardiovascular diseases. Hirudin is widely used in the prevention and treatment of cardiovascular diseases and cancers. In this study, we examined the potential role(s) and mechanism of hirudin in angiotensin II (Ang II)-induced myocardial fibrosis.Material/MethodsThe viability of myocardial fibroblasts, and reactive oxygen species (ROS) rates were measured respectively using cell counting kit-8 (CCK-8) and flow cytometry. Malondialdehyde (MDA) content, the activities of lactate dehydrogenase (LDH), and superoxide dismutase (SOD) were detected by the respective kits. The mRNA and protein levels of fibrosis-related factors were separately assessed by qRT-PCR and western blot.ResultsOur data revealed that hirudin suppressed the viability of myocardial fibroblasts, and that it relieved the proliferation induced by Ang II in a dose-dependent manner. We also found that hirudin reduced ROS production, LDH activity, and MDA content; however, it enhanced SOD activity. Moreover, while hirudin significantly downregulated the levels of matrix metalloproteinase-2 (MMP-2), MMP-9, fibronectin (FN), transforming growth factor beta 1 (TGF-β1), collagen-I (COL-I), and COL-III, it upregulated the expression level of tissue inhibitor of metalloproteinases-2 (TIMP-2). Furthermore, phosphorylated extracellular signal-regulated kinase1/2 (p-ERK1/2) was decreased by hirudin, compared to the Ang-II group.ConclusionsHirudin depressed Ang II-induced myocardial fibroblasts via inhibiting oxidative stress, regulating fibrosis-related factors, and repressing the ERK1/2 pathway.

The Association of Ascorbic Acid, Deferoxamine and N-Acetylcysteine Improves Cardiac Fibroblast Viability and Cellular Function Associated with Tissue Repair Damaged by Simulated Ischemia/Reperfusion

“…Among these, type I collagen is widely expressed in cardiac tissue of mammals and forms thick and stiff fibers [26]. Angiotensin II is a peptide known to elicit cardiac fibrosis [51,52] by stimulating CF collagen production and secretion [53,54]. In our CF in vitro model, sI/R prevented the increase of pro-collagen I synthesis triggered by angiotensin II and did not induce pro-collagen I production by itself.…”

Pharmacological Activities and Mechanisms of Hirudin and Its Derivatives - A Review

“…Besides, the reactive oxygen species (ROS) production, lactate dehydrogenase (LDH) activity, and MDA content were reduced by hirudin, while the SOD activity was enhanced. Moreover, the levels of several fibrosis-related factors such as matrix metalloproteinase-2 (MMP-2), MMP-9, FN, TGF-β 1 , COL-I, and COL-III were significantly down-regulated, but the expression level of tissue inhibitor of metalloproteinases-2 (TIMP-2) was upregulated (Yu et al, 2018).…”

Roles of Biomarkers in Myocardial Fibrosis

“…TIMP1 mediates the interaction between fibroblast membrane protein CD63 and integrin β1, which is widely expressed in various cells, thus triggering fibrosis [ 52 - 55 ]. At the cellular level, hirudin reverses Ang II-induced fibrosis by elevating TIMP-2 expression [ 56 ]. TIMP3 is the only TIMP that can inhibit the activity of TNF-α-converting enzyme and may have a protective effect against fibrosis by upregulating cytokines involved in myofibroblast activation and immunity [ 57 ].…”