Application of specific excitation Mas receptor in medicine for preventing and treating hepatic ischemia-reperfusion injury

Abstract

The invention relates to an application of a specific excitation Mas receptor in a medicine for preventing and treating hepatic ischemia-reperfusion injury. According to the invention, a G protein coupled receptor Mas coded by a protooncogene Mas1 is focused, a 70% warm ischemia reperfusion model of the liver is constructed by clamping left and middle leaf vessels of the liver by a vascular clamp after a mouse is laparotomy, and the condition that the liver injury can be obviously alleviated by activating the Mas receptor and the liver injury can be aggravated by knocking out the Mas receptor is fully proved on an in-vivo level; the invention provides an important early-stage data basis for research and development of new drugs based on specific activation of the Mas receptor, and is expected to provide an important animal experiment basis for development of new drugs aiming at the Mas receptor in a later stage.

Description

Technical Field

[0001] The invention belongs to the field of liver ischemia-reperfusion injury, and particularly relates to the application of specific stimulatory Mas receptors in drugs for preventing and treating liver ischemia-reperfusion injury. Background Art

[0002] Liver ischemia-reperfusion injury (LIRI) is the most common unavoidable adverse event after liver transplantation, which seriously affects clinical outcomes. Sustained immune inflammatory response is an important driving force for its progression. Host innate immune activation is the core of LIRI, including the infiltration of multiple inflammatory cells such as neutrophils and macrophages, which ultimately transforms the liver's immune tolerance microenvironment into an inflammatory organ. In this process, neutrophils are recruited to the site of injury as innate effector immune cells to aggravate tissue damage, especially aging neutrophils, which have been considered to be a more pro-inflammatory subpopulation. At the same time, activated macrophages also play an important pathogenic role by secreting a large number of pro-inflammatory factors. Increased infiltration of liver macrophages and neutrophils has been considered to be an established biomarker of the severity of LIRI. Previous studies on inflammatory cells have focused on reducing the infiltration and accumulation of immune cells or avoiding the production of neutrophil extracellular traps, but all have certain limitations.

[0003] Although they are considered to be the main immune effector cells of sterile inflammation, more and more studies have shown that immune cells such as macrophages and neutrophils interact with each other and play an important role in maintaining tissue homeostasis and injury repair, suggesting that endogenous cell interactions may be a more promising direction in the future. Among them, the phagocytosis of apoptotic cells, including neutrophils, by macrophages is called "efferocytosis", which is a key process for macrophage polarization to initiate the inflammatory repair stage. Many studies from the cardiovascular, liver, and kidney fields have demonstrated the important role of macrophage efferocytosis in inflammatory repair and maintaining tissue homeostasis. Therefore, exploring the key molecules and mechanisms of the macrophage efferocytosis process and exploring the important endogenous regulatory factors that maintain macrophage functional homeostasis may help to discover new therapeutic targets for LIRI.

[0004] Mas receptor is a G protein-coupled receptor encoded by the proto-oncogene Mas1, which is widely expressed in various tissues of humans and mice. The natural ligand of Mas is angiotensin-1-7 (ANG-(1-7)), which is an important effector molecule of the "anti-inflammatory" renin-angiotensin system. Activating Mas has multiple beneficial effects in the liver, such as anti-inflammation, improvement of glucose and lipid metabolism disorders, and anti-fibrosis, but many of the mechanisms are still unclear. Summary of the invention

[0005] The present invention provides application of specific agonist Mas receptor in medicine for preventing and treating liver ischemia-reperfusion injury, aiming at the technical problem that existing medicines generally have limitations.

[0006] The invention provides application of a specific stimulatory Mas receptor in a drug for preventing and treating liver ischemia-reperfusion injury.

[0007] Furthermore, the drug includes Mas receptor-specific agonist AVE0991.

[0008] Furthermore, the drug comprises the Mas ligand ANG-(1-7).

[0009] Furthermore, the drug also contains pharmaceutically acceptable carriers and / or excipients.

[0010] Furthermore, the pharmaceutically acceptable carrier and / or excipient includes at least one of a diluent, a binder, a surfactant, a humectant, an adsorption carrier, a lubricant, a filler, and a disintegrant.

[0011] In the present invention, myeloid cell-specific Mas1 gene knockout mice (LysM cre Mas1 f / f ) and control mice (Mas1 f / f ) After laparotomy, a 70% warm ischemia-reperfusion model of the liver was constructed by clamping the common trunk of the left and middle lobe vessels with a vascular clamp. The results showed that under the same modeling conditions, myeloid cell-specific Mas1 gene knockout mice showed more severe liver damage and intrahepatic inflammatory response compared with control mice. In addition, specific activation of Mas receptors before LIRI modeling of wild-type mice can significantly reduce liver damage caused by ischemia-reperfusion. The present invention confirms that Mas receptor activation can play a protective role against liver ischemia-reperfusion injury by enhancing macrophage efferocytosis. Based on the current research data, Mas receptors can be used as a highly potential therapeutic target for the prevention and treatment of liver ischemia-reperfusion injury.

[0012] Beneficial Effects

[0013] The present invention focuses on the G protein-coupled receptor Mas encoded by the proto-oncogene Mas1. By opening the abdomen of mice and clamping the common trunk of the left and middle lobe vessels with vascular clamps, a 70% warm ischemia-reperfusion model of the liver was constructed. It was fully confirmed at the in vivo level that activating the Mas receptor can significantly reduce liver damage, while knocking out the Mas receptor can aggravate liver damage, and the mechanism is explained to be related to macrophage efferocytosis, which provides an important preliminary data basis for the development of new drugs based on the specific activation of the Mas receptor, and is expected to provide an important animal experimental basis for the later development of new drugs targeting the Mas receptor. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 AF shows that myeloid cell-specific Mas knockout leads to massive accumulation of senescent neutrophils.

[0015] Figure 2 AE is a figure showing that the Mas receptor-specific agonist AVE0991 significantly alleviates liver ischemia-reperfusion injury by enhancing macrophage efferocytosis.

[0016] Figure 3 AF is the Mas ligand Ang-(1-7) which significantly reduces liver ischemia-reperfusion injury by activating Mas receptors and enhancing macrophage efferocytosis.

[0017] Figure 4 AD shows that Mas ligand Ang-(1-7) promotes tissue repair after liver ischemia-reperfusion injury by activating Mas receptors to enhance macrophage efferocytosis. DETAILED DESCRIPTION

[0018] The present invention will be further described below in conjunction with specific embodiments. It should be understood that these embodiments are only used to illustrate the present invention and are not intended to limit the scope of the present invention. In addition, it should be understood that after reading the content taught by the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms fall within the scope limited by the appended claims of the application equally.

[0019] Example 1

[0020] 1. Experimental Methods

[0021] (1) Construction of myeloid cell-specific Mas knockout gene Mas1 using the Loxp-Cre recombinase system fl / fl LysM-Cre mice and littermate controls Mas1 fl / fl A LIRI model was established in mice (6-8 weeks, male). 24 h after reperfusion, liver tissue (HE staining) and serological indicators (ALT) were detected in mice. mIHC (F4 / 80 / MerTK / Mas / Ly6G / CXCR4) and in vivo imaging (WGA / Ly6G / CXCR4 / F4 / 80) were used to observe the inter-group differences in macrophage efferocytosis and senescent neutrophils in the mouse liver to clarify that Mas receptor participates in tissue damage repair by affecting macrophage efferocytosis.

[0022] (2) LIRI model was established in 6-8 week old male wild-type C57BL / 6 mice. The control group was intraperitoneally injected with solvent control 24 h and 3 h before modeling, while the experimental group was intraperitoneally injected with Mas receptor specific agonist AVE0991 24 h and 3 h before modeling to activate Mas receptor. 24 h after reperfusion, liver tissue (HE staining) and serological indicators (ALT) were detected in mice. mIHC (F4 / 80 / MerTK / Mas / Ly6G / CXCR4) was used to observe the differences in macrophage efferocytosis and senescent neutrophils in the liver of mice, so as to clarify the effect and feasibility of targeted intervention of Mas receptor to upregulate macrophage efferocytosis, reduce inflammatory damage and promote tissue repair.

[0023] (3) LIRI model was established in 6-8 week old male wild-type C57BL / 6 mice. The control group was intraperitoneally injected with solvent control 72 h, 48 h, 24 h before modeling and 6 h after reperfusion. The experimental group was intraperitoneally injected with Mas ligand Ang-(1-7) 72 h, 48 h, 24 h before modeling and 6 h after reperfusion to activate Mas receptor. Liver tissue (HE staining) and serological indicators (ALT) were measured 1 d and 3 d after reperfusion. mIHC (F4 / 80 / MerTK / Mas / Ly6G / CXCR4) was used to observe the differences in macrophage efferocytosis and senescent neutrophils in the liver of mice, so as to clarify the effect and feasibility of targeted intervention of Mas receptor to upregulate macrophage efferocytosis, alleviate inflammatory damage and promote tissue repair.

[0024] 2. Experimental Results

[0025] Figure 1 AF showed that compared with the control group, myeloid cell-specific Mas1 gene knockout could significantly aggravate the degree of liver ischemia-reperfusion injury and cause a large number of senescent neutrophils to accumulate in the necrotic area.

[0026] Figure 2 AE showed that the Mas receptor-specific agonist AVE0991 could significantly alleviate liver ischemia-reperfusion injury by enhancing macrophage efferocytosis.

[0027] Figure 3 AF showed that Mas ligand Ang-(1-7) significantly alleviated liver ischemia-reperfusion injury by activating Mas receptor and enhancing macrophage efferocytosis.

[0028] Figure 4 AD showed that Mas ligand Ang-(1-7) promoted tissue repair after liver ischemia-reperfusion injury by activating Mas receptors and enhancing macrophage efferocytosis.

Claims

1. Application of drugs that specifically stimulate Mas receptors in the prevention and treatment of liver ischemia-reperfusion injury.

2. The use according to claim 1, characterized in that: The drug includes the Mas receptor specific agonist AVE0991.

3. The use according to claim 1, characterized in that: The drugs include the Mas ligand ANG-(1-7).

4. The use according to claim 1, characterized in that: The medicine further comprises a pharmaceutically acceptable carrier and / or excipient.

5. The use according to claim 4, characterized in that: The pharmaceutically acceptable carrier and / or excipient includes at least one of a diluent, a binder, a surfactant, a humectant, an adsorption carrier, a lubricant, a filler, and a disintegrant.

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