Application of KLF4 in preparation of drug for treating drug-induced liver injury

Abstract

The invention relates to application of KLF4 in preparation of a drug for treating drug-induced liver injury. On an APAP-induced mouse acute liver injury model, it is fully proved that activation of the KLF4 can significantly alleviate liver injury and liver tissue inflammatory response, and knockout of the KLF4 can significantly aggravate liver injury and liver tissue inflammatory response, so that an important earlier-stage data basis is provided for research and development of new drugs based on the specific activation transcription factor KLF4; and important help is expected to be provided for development of new drugs aiming at the transcription factor KLF4 in the later period.

Description

Technical Field

[0001] The present invention belongs to the field of drug-induced liver injury, and particularly relates to the application of KLF4 in the preparation of a drug for treating drug-induced liver injury. Background Art

[0002] Drug-induced liver injury (DILI) is the most common adverse drug reaction, with high morbidity and mortality, and is also an important obstacle to the development and marketing of new drugs. Among them, liver injury caused by acetaminophen (APAP) is a typical hepatocellular injury-type DILI. Due to the availability of mouse models highly relevant to human pathophysiology, APAP is also the most widely studied hepatotoxic drug. In recent years, the study of Krüppel-like Factor 4 (KLF4) in the field of inflammation has attracted widespread attention. KLF4 is a transcription factor characterized by a zinc finger protein structure. Increasing evidence shows that KLF4 plays a key role in the pathogenesis of various inflammatory diseases. At present, there are no reports on the study of KLF4 in drug-induced liver injury. Summary of the invention

[0003] The present invention provides the use of KLF4 in preparing a drug for treating drug-induced liver injury, aiming at the technical problem that existing drug intervention means are insufficient.

[0004] The present invention provides application of KLF4 in preparing a medicine for treating drug-induced liver injury.

[0005] Furthermore, activating the KLF4 reduces liver damage and inflammatory response in liver tissue.

[0006] Furthermore, the drug-induced liver injury includes acetaminophen-induced drug-induced liver injury.

[0007] Furthermore, the drug also contains pharmaceutically acceptable carriers and / or excipients.

[0008] 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.

[0009] The present invention is to treat myeloid cell-specific Klf4 gene knockout mice (LysM cre Klf4 f / f ) and control mice (Klf4 f / f) were intraperitoneally injected with APAP to construct an in vivo research model of DILI. The results showed that with the same dose of APAP, more severe hepatocellular damage and intrahepatic inflammatory response were observed in myeloid cell-specific Klf4 gene knockout mice compared with the control group mice. Before APAP modeling in wild-type (WT) mice, the use of the KLF4 small molecule agonist APTO-253 to specifically activate the transcription factor KLF4 significantly reduced APAP-induced liver damage.

[0010] Beneficial Effects

[0011] The present invention has fully demonstrated in the APAP-induced acute liver injury model in mice that activating KLF4 can significantly reduce liver injury and inflammatory response in liver tissue, while knocking out KLF4 can significantly aggravate liver injury and inflammatory response in liver tissue, providing an important preliminary data basis for the development of new drugs based on the specific activation of the transcription factor KLF4, and is expected to provide important assistance for the later development of new drugs targeting the transcription factor KLF4. BRIEF DESCRIPTION OF THE DRAWINGS

[0012] Figure 1 AC, Figure 2 AC showed that knocking out KLF4 transcription factor 24 hours later could significantly aggravate APAP-induced acute liver injury.

[0013] Figure 3 AB showed that knocking out KLF4 transcription factor 48 hours later aggravated APAP-induced acute liver injury.

[0014] Figure 4 AB showed that activation of KLF4 transcription factor could significantly alleviate APAP-induced acute liver injury. DETAILED DESCRIPTION

[0015] 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.

[0016] Example 1

[0017] 1 Experimental animals and methods

[0018] 1.1a. Construction of disease model with transgenic mice: ① Transgenic mice: Klf4 in C57BL / 6 background f / f With LysM cre Myeloid cell-specific Klf4 knockout mice (LysM cre Klf4 f / f②Disease model: administration of Klf4 f / f Mouse and LysM cre Klf4 f / f Mice were intraperitoneally injected with APAP (300 mg / kg, 24h-48h) to establish an APAP-induced liver injury model.

[0019] b. Experimental design: ① Collect blood from the eye sockets and separate serum to detect ALT, AST, and ALP in serum to preliminarily evaluate the degree of liver damage. ② Prepare paraffin sections of liver tissue and perform H&E staining, MPO staining, F4 / 80 staining, and TUNEL staining to evaluate intrahepatic inflammation and necrosis; ③ Extract liver tissue protein and detect changes in CYP2E1 expression by WB; ④ Collect fresh liver tissue specimens for BulkRNA-seq sequencing to detect LysM cre Klf4 f / f Mouse and Klf4 f / f Differential expression of mRNA in mouse disease models.

[0020] 1.2a. Disease model and drug intervention: ① Mice: Wild-type mice with C57BL / 6 background. ② Disease model: Small molecule KLF4-specific agonist (APTO-253) or solvent control was administered before intraperitoneal injection of APAP to induce acute liver injury model in mice.

[0021] b. Experimental design: ① Blood was collected from the eye sockets and serum was separated to detect ALT and AST in the serum to preliminarily assess the degree of liver damage. ② Paraffin sections of liver tissue were prepared and H&E staining was performed to assess the necrotic changes in liver tissue.

[0022] 2 Experimental results

[0023] 2.1 Figure 1 The results showed that knocking out KLF4 transcription factor 24 hours later could significantly aggravate liver damage (H&E staining, increased serum ALT, AST, and ALP levels) and inflammatory response (MPO and F4 / 80 staining).

[0024] 2.2 Figure 2 It was shown that 24 hours after knocking out the KLF4 transcription factor, the inflammatory pathway in liver tissue was upregulated (Bulk RNA-seq sequencing).

[0025] 2.3 Figure 3 The results showed that knocking out KLF4 transcription factor could aggravate liver damage (H&E staining, increased serum ALT, AST, and ALP levels) after 48 hours.

[0026] 2.4 Figure 4 The results showed that specific activation of KLF4 transcription factor could significantly reduce liver damage (H&E staining, decreased serum ALT and AST levels).

Claims

1. Application of KLF4 in the preparation of drugs for the treatment of drug-induced liver injury.

2. The use according to claim 1, characterized in that: Activating KLF4 reduces liver damage and inflammatory response in liver tissue.

3. The use according to claim 1, characterized in that: The drug-induced liver injury includes acetaminophen-induced drug-induced liver injury.

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 wetting agent, an adsorption carrier, a lubricant, a filler, and a disintegrant.

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