Application of uric acid in preparation of medicines for treating liver injury and platelet inhibition medicines

Abstract

The invention relates to the technical field of medicines, in particular to application of uric acid in preparation of a medicine for treating liver injury and a medicine for inhibiting platelets during liver injury. The invention relates to an application of uric acid in the aspects of treating liver injury and inhibiting platelet activation, which indicates that uric acid (especially exogenous uric acid) can be used for preparing the medicines for treating liver injury and platelet activation, and provides technical enlightenment for preparing the medicines for treating liver injury and liver cirrhosis hypercoagulable state. The invention discloses that exogenous uric acid has a treatment effect on acute liver injury induced by thioacetamide (TAA). The exogenous uric acid has a treatment effect on chronic liver injury caused by cholestasis. The mechanism is related to reduction of oxidative stress and inflammatory response and further reduction of cell senescence. The condition of a liver cirrhosis patient is related to the platelet activation state, exogenous uric acid inhibits TAA and platelet activation in a common bile duct ligation (BDL) rat model, and it is prompted that the liver disease and the liver cirrhosis hypercoagulable state can be treated.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, specifically the application of uric acid in the preparation of drugs for treating liver damage and platelet-inhibiting drugs. Background Technology

[0002] Acute liver injury is liver damage induced by various etiologies. If left untreated, it can develop into acute liver failure, seriously endangering the patient's life. Clinical treatment faces the challenge of comprehensively addressing all these issues, and the medications used in clinical treatment can also cause various adverse reactions. Chronic liver injury causes nearly 2 million deaths annually. Therefore, there is an urgent need to explore more reliable treatment measures. Currently, several programs exist for the prevention and treatment of liver disease, but they can also cause various adverse reactions. Therefore, further development of corresponding drugs is necessary.

[0003] Uric acid, a product of purine nucleotide metabolism in the body, is a natural antioxidant. Most previous research on uric acid has focused on the relationship between serum uric acid levels and diseases. It was previously believed that high serum uric acid was associated with metabolic diseases, cardiovascular diseases, and kidney diseases. However, recent studies have found that lowering serum uric acid has not shown a clear clinical therapeutic effect in these diseases, suggesting that there may be no causal relationship between hyperuricemia and these diseases. The relationship between serum uric acid and liver disease is also unclear. Serum uric acid levels may change when liver function is impaired. Previously, serum uric acid levels were considered an indicator of liver damage. However, recent studies have shown that uric acid levels may decrease as liver function deteriorates, and this is associated with poor prognosis. Most scholars believe that high serum uric acid is an independent risk factor for non-alcoholic fatty liver disease / metabolic fatty liver disease. However, some studies have found a negative correlation between serum uric acid levels and fibrosis stage in NAFLD patients.

[0004] In summary, despite years of research on uric acid, a unified consensus on its precise role in disease remains lacking, particularly regarding its underestimated medicinal value. Uric acid possesses strong antioxidant capabilities, capable of scavenging approximately 60% of free radicals in the body. Most commonly used Parkinson's disease medications currently available in clinical practice can increase serum uric acid levels. A moderate increase in uric acid in the short term is a temporary physiological fluctuation, its effect possibly related to antioxidant activity; however, long-term hyperuricemia is a persistent pathological state that can lead to gout, kidney damage, and other diseases, and the two should not be equated. Therefore, the applicant is exploring whether small amounts of uric acid also have a therapeutic effect on liver disease.

[0005] Under normal circumstances, platelets contain various cytokines that play a crucial role in maintaining liver homeostasis. Recent studies have found that patients with liver disease often exhibit abnormal platelet function and activation. Activated platelets play a promoting role in the development and progression of liver fibrosis. In patients with cirrhosis, changes in vascular endothelial structure and impaired function promote platelet adhesion and aggregation, leading to a hypercoagulable state. Therefore, recent studies suggest that antiplatelet drugs have potential therapeutic effects on liver diseases and liver fibrosis. However, safely and effectively using antiplatelet drugs without increasing the risk of bleeding remains a significant challenge, thus requiring further drug development. Summary of the Invention

[0006] To address the aforementioned problems, this invention provides the application of uric acid in the preparation of drugs for treating liver injury and platelet-inhibiting drugs, as detailed below: Applications of uric acid in the preparation of drugs for treating liver damage.

[0007] Furthermore, the aforementioned drug for treating liver injury is a drug for treating TAA-induced liver injury.

[0008] Furthermore, the aforementioned drug for treating liver injury is a drug for treating BDL-induced liver injury.

[0009] Furthermore, the aforementioned drug for treating liver damage is a drug that reduces liver fibrosis.

[0010] Furthermore, the drug for treating liver injury is a drug that reduces liver inflammation and oxidative stress. Preferably, the drug that reduces liver inflammation and oxidative stress is a drug that lowers lactate dehydrogenase (LDH) and serum creatinine.

[0011] Furthermore, the aforementioned liver injury treatment drug is a drug that reduces inflammation and aging markers in BDL liver injury.

[0012] Furthermore, the aforementioned drug for treating liver damage is a drug that reduces spleen enlargement.

[0013] Applications of uric acid in the preparation of platelet-inhibiting drugs.

[0014] Furthermore, the platelet-inhibiting drug is a drug that inhibits platelet activation.

[0015] Furthermore, the platelet-inhibiting drug is a drug that inhibits platelet activation in patients with liver damage.

[0016] Furthermore, the platelet-inhibiting drug is a drug that reduces platelet inflammation in patients with liver damage.

[0017] Furthermore, the platelet-inhibiting drug is a medication used to treat liver diseases and hypercoagulable states associated with cirrhosis.

[0018] Compared with the prior art, the technical effects of this invention are reflected in: This invention relates to the application of uric acid in the treatment of liver damage and the inhibition of platelet activation, demonstrating that uric acid (especially exogenous uric acid) can be used to prepare drugs for treating liver damage and platelet inhibition, providing technical inspiration for the preparation of drugs for liver damage and hypercoagulable states in cirrhosis. Its specific effects on liver damage are as follows: ① TAA induces liver dysfunction in rats, and exogenous uric acid alleviates TAA-induced liver damage; ② TAA induces changes such as cell necrosis and inflammatory cell infiltration, and uric acid alleviates liver damage; ③ TAA induces increased expression of BAX, p21, and ANXA1 in the liver, and uric acid reduces the increase in the expression of these proteins; ④ TAA induces the degree of liver fibrosis, and uric acid reduces the degree of fibrosis; ⑤ TAA induces the expression of IL-6 and CD62p in the liver, and uric acid inhibits this response; ⑥ TAA induces increased serum 3-NT, LDH, and serum creatinine, and uric acid reduces LDH and serum creatinine; ⑦ TAA induces hepatocyte damage, enhanced oxidative stress response, and increased expression of ANXA1 and p21. Exogenous uric acid alleviates the above changes; ⑧ BDL leads to an increase in liver function indicators in rats, and exogenous uric acid alleviates liver damage; ⑨ BDL leads to an increase in the expression of p21 and IL-1β in the liver, and uric acid alleviates the above changes; ⑩ BDL leads to spleen enlargement in rats, and exogenous uric acid alleviates this response. Its inhibitory effects on platelet activation are as follows: ① Uric acid inhibits hippocampal platelet accumulation and increases CD62P protein expression in the TAA model; ② Uric acid inhibits platelet activation; ③ Uric acid reduces spleen size in the BDL model; ④ Uric acid alleviates platelet inflammation in the BDL model. Attached Figure Description

[0019] Figure 1 This is the result of liver function index testing for exogenous uric acid reduction.

[0020] Figure 2 The HE staining results indicate that exogenous uric acid reduces liver damage.

[0021] Figure 3 The results show that exogenous uric acid reduces cell apoptosis and the expression of p21 and ANXA1.

[0022] Figure 4 The test results indicate that exogenous uric acid reduces liver fibrosis.

[0023] Figure 5 The results indicate that exogenous uric acid reduces liver inflammation and oxidative stress.

[0024] Figure 6 The study used a cell model to discover that uric acid reduces TAA-induced hepatocyte damage in rats.

[0025] Figure 7This is a graph showing the results of blood biochemistry indicators indicating that exogenous uric acid has a protective effect against BDL liver damage.

[0026] Figure 8 This is the result of testing for inflammatory and aging indicators that exogenous uric acid can reduce BDL liver damage.

[0027] Figure 9 This is the result of detecting spleen volume induced by exogenous uric acid reducing BDL.

[0028] Figure 10 The results show increased platelet accumulation in the hippocampus and elevated CD62P protein expression in the uric acid-inhibited TAA model.

[0029] Figure 11 This is the result of a test showing that uric acid inhibits platelet activation.

[0030] Figure 12 The blood routine test results indicated that uric acid inhibited platelet activation in the BDL model.

[0031] Figure 13 This is the spleen detection result of the BDL model, which reduces exogenous uric acid levels.

[0032] Figure 14 This is the result of platelet inflammation detection in the BDL model, which shows a decrease in exogenous uric acid. Detailed Implementation

[0033] The technical solution of the present invention will be further defined below with reference to specific embodiments, but the scope of protection is not limited to the description made.

[0034] Research on the use of uric acid in the preparation of drugs for treating liver injury Example 1: Uric acid alleviates TAA-induced liver injury in rats 1. Model and Grouping An acute HE model was established by intraperitoneal injection of TAA 200 mg / kg into rats for 3 consecutive days.

[0035] SD rats were divided into three groups: WT control group (Control), WT model group (TAA), and WT treatment group (TAA+UA). ANXA1 rats were divided into three groups: AN- / - control group (AN- / - Control), AN- / - model group (AN- / -TAA), and AN- / - treatment group (AN- / -TAA+UA).

[0036] Control group: Intraperitoneal injection of 2 ml / kg of normal saline containing the same amount of TAA as the HE model group; Model group: TAA was injected intraperitoneally at a concentration of 100 mg / ml for 3 consecutive days at a concentration of 200 mg / kg / d. Treatment group: TAA was administered intraperitoneally at a dose of 200 mg / kg / day for 3 consecutive days, with uric acid 200 mg / kg administered intraperitoneally on the last day.

[0037] 2. Exogenous uric acid reduces liver function indicators Method: Kit.

[0038] Results: See details below. Figure 1 TAA-induced liver dysfunction in rats was alleviated by exogenous uric acid.

[0039] 3. HE staining results suggest that exogenous uric acid reduces liver damage. Method: HE staining.

[0040] Results: See details below. Figure 2 TAA induces changes such as cell necrosis and inflammatory cell infiltration, while uric acid reduces liver damage.

[0041] 4. Exogenous uric acid reduces apoptosis and p21, ANXA1 expression. Method: Western blot.

[0042] Results: See details below. Figure 3 TAA induces increased expression of BAX, p21, and ANXA1 in the liver, while decreased uric acid leads to increased expression of these proteins.

[0043] 5. Exogenous uric acid reduces liver fibrosis Method: Masson staining of the liver.

[0044] Results: See details below. Figure 4 TAAs induce liver fibrosis, while uric acid reduces the degree of fibrosis.

[0045] 6. Exogenous uric acid reduces liver inflammation and oxidative stress. Methods: Western blot and 3-NT, LDH and serum creatinine kits.

[0046] Results: See details below. Figure 5 TAA induces the expression of IL-6 and CD62p in the liver, while uric acid inhibits this response. TAA induces increased serum 3-NT and LDH, as well as serum creatinine, while uric acid reduces LDH and serum creatinine.

[0047] Example 2: Using a cell model, it was found that uric acid alleviates TAA-induced hepatocyte damage in rats. Methods: LDH kit. DHE staining to detect ROS. Immunofluorescence to observe ANXA1. Western blotting to detect P21.

[0048] Results: See details below. Figure 6TAA induces hepatocyte damage, enhances oxidative stress, and increases the expression of ANXA1 and p21. Exogenous uric acid alleviates these changes.

[0049] Example 3: Protective effect of exogenous uric acid on a BDL rat model of liver injury. 1. Modeling and Grouping This experiment was divided into three groups: Sham group: except that the common bile duct was not ligated, the rats were treated the same as the BDL group and injected with the same amount of physiological saline; BDL+NS group: the common bile duct was ligated for four weeks to establish a chronic HE model; BDL+UA group: exogenous uric acid (150 mg / kg, concentration of 50 mg / mL) was injected intraperitoneally twice a week for a total of four times in the third week.

[0050] 2. Blood biochemical indicators suggest that exogenous uric acid has a protective effect against BDL liver damage. Method: Kit.

[0051] Results: See details below. Figure 7 BDL led to an increase in liver function indicators in rats, while exogenous uric acid reduced liver damage.

[0052] 3. Exogenous uric acid reduces inflammation and aging markers in BDL-induced liver damage. Method: Western blot.

[0053] Results: See details below. Figure 8 BDL leads to increased expression of p21 and IL-1β in the liver, while uric acid reduces these changes.

[0054] 4. Exogenous uric acid reduces BDL-induced spleen volume. Method: Direct measurement.

[0055] Results: See details below. Figure 9 BDL caused spleen enlargement in rats, and exogenous uric acid alleviated this response.

[0056] Research findings on the use of uric acid in the preparation of drugs for treating liver injury: 200 mg / kg exogenous uric acid has a therapeutic effect on TAA-induced acute liver injury. 150 mg / kg exogenous uric acid has a therapeutic effect on BDL-induced chronic liver injury. The mechanism is related to reducing oxidative stress and inflammatory responses, thereby alleviating cellular senescence.

[0057] Research on the use of uric acid in the preparation of platelet-inhibiting drugs Example 4: TAA Experimental Section Modeling: In this study, an acute liver injury rat model was established by intraperitoneal injection of TAA (200 mg / kg) for three consecutive days. Rats were divided into three groups: the control group (Control) received intraperitoneal injection of the same volume of physiological saline (2 mL / kg) as the HE model group; the model group (TAA) and the treatment group (TAA+UA) received intraperitoneal injection of TAA (200 mg / kg, concentration 100 mg / mL) for three consecutive days. On the third day, the treatment group also received intraperitoneal injection of uric acid (200 mg / kg, concentration 100 mg / mL), while the model group received an equal volume of physiological saline.

[0058] 1. Uric acid inhibits increased hippocampal platelet accumulation and CD62P protein expression in the TAA model. Methods: Immunofluorescence staining and Western blot.

[0059] Results: See details below. Figure 10 Immunofluorescence showed increased CD61 accumulation in the hippocampus of the TAA group, and Western blot results showed increased CD62p (p=0.0098). Uric acid treatment alleviated these changes (p=0.0089).

[0060] 2. Uric acid inhibits platelet activation. The centrifugation conditions for platelet-rich plasma (PRP) were: 200 g centrifugation for 10 min at room temperature.

[0061] Methods: Flow cytometry and Western blot.

[0062] Results: See details below. Figure 11 Flow cytometry results showed that the platelet content in platelet-rich serum (PRP) was approximately 97.04%. Western blot results showed that CD62P expression in PRP of the TAA model group was increased compared with that of the control group (p=0.0003), while UA inhibited CD62P expression (p=0.0013).

[0063] Example 5 BDL Experiment Section Modeling: Establishment of the BDL rat model First, rats were anesthetized with sevoflurane and fixed on the operating table. Anesthesia was maintained through continuous inhalation of sevoflurane. The surgical area was shaved and disinfected. A 2-3 cm incision was made along the midline of the abdomen, and the skin and abdominal wall muscles were dissected layer by layer to expose the abdominal cavity. Next, the abdominal cavity was opened to expose the common bile duct. Double ligation of the common bile duct was performed at two different locations using 6-0 sutures. The common bile duct was then cut between the ligation points to ensure complete cessation of bile flow. Finally, the surgical area was carefully examined to confirm the absence of bleeding and other abnormalities. The abdominal wall muscles and skin were sutured layer by layer, and the rats were placed in a warm environment until fully recovered.

[0064] 1. Blood routine test results indicated that uric acid inhibited platelet activation in the BDL model. Method: Complete blood count (CBC) test.

[0065] Results: See details below. Figure 12 The platelet-to-large ratio (P-LCR) is an indicator of platelet activation. In the BDL group, white blood cell count (p<0.0001), platelet count (p=0.0456), platelet-to-hematocrit (PCT) (p=0.0058), and P-LCR (p=0.0037) were higher than in the Sham group. After the application of exogenous UA, white blood cell count (p<0.0001), platelet count (ns), PCT (p=0.0213), and P-LCR (p=0.0104) decreased.

[0066] 2. Spleen in the BDL model with reduced exogenous uric acid Method: See details for specific results. Figure 13 Hypersplenism can lead to platelet activation; therefore, this study measures spleen size to indirectly reflect the platelet activation status.

[0067] Results: The spleen area in the BDL group was larger than that in the Sham group (p=0.0001), and decreased after UA intervention (p=0.0092).

[0068] 3. Exogenous uric acid reduces platelet inflammation in the BDL model. Method: Western blot. Results: See details below. Figure 14 In the BDL group, IL-1β protein expression (p=0.001) was increased in PRP compared to the Sham group, while IL-1β protein expression (p=0.0044) decreased after the application of exogenous UA.

[0069] Research findings on the use of uric acid in the preparation of platelet-inhibiting drugs: Liver disease leads to platelet activation. This project used TAA and BDL rat models to find that exogenous uric acid can inhibit platelet activation in the models.

[0070] The condition of patients with cirrhosis is correlated with platelet activation status. Exogenous uric acid inhibits platelet activation in rat models of TAA and BDL, suggesting a potential therapeutic effect on liver disease and the hypercoagulable state of cirrhosis.

[0071] Finally, it should be noted that the above embodiments are merely representative examples of the present invention. Obviously, the technical solution of the present invention is not limited to the above embodiments, and many variations are possible. All variations that can be directly derived or conceived by those skilled in the art from the content disclosed in this invention should be considered within the scope of protection of this invention.

Claims

1. Application of uric acid in the preparation of drugs for treating liver damage.

2. The application according to claim 1, characterized in that, The aforementioned drug for treating liver injury is a drug for treating TAA-induced liver injury.

3. The application according to claim 1, characterized in that, The aforementioned drug for treating liver injury is a drug for treating BDL-induced liver injury.

4. The application according to claim 1, characterized in that, The aforementioned medication for treating liver damage is a drug that reduces liver fibrosis.

5. The application according to claim 1, characterized in that, The aforementioned medication for treating liver damage is designed to reduce liver inflammation and oxidative stress.

6. The application according to claim 5, characterized in that, The drugs mentioned that reduce liver inflammation and oxidative stress are those that lower lactate dehydrogenase (LDH) and serum creatinine.

7. The application according to claim 1, characterized in that, The aforementioned liver injury treatment drug is designed to reduce inflammation and aging markers associated with BDL liver injury.

8. The application according to claim 1, characterized in that, The aforementioned medication for treating liver damage is one that reduces spleen enlargement.

9. Application of uric acid in the preparation of platelet-inhibiting drugs.

10. The application according to claim 9, characterized in that, The platelet-inhibiting drug is a drug that inhibits platelet activation in patients with liver damage.

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