New Applications of Adenosylcobalamin

Abstract

The present invention belongs to the technical field of biological medicines, and relates to a new application of adenosylcobalamin, specifically, the application of adenosylcobalamin in the preparation of a drug for treating or preventing chronic pancreatitis. Adenosylcobalamin can inhibit macrophage M2 polarization, reduce pancreatic fibrosis caused by chronic pancreatitis, and reduce pancreatic tissue changes caused by chronic pancreatitis. The present invention provides a basis for drug screening for chronic pancreatitis.

Description

Technical Field

[0001] The invention belongs to the technical field of biological medicines, and in particular relates to a new application of adenosylcobalamin. Background Art

[0002] Chronic pancreatitis (CP) is a progressive fibroinflammatory disease with permanent damage to the pancreatic structure. The main pathological manifestations of CP are pancreatic acinar atrophy, tissue fibrosis or calcification, focal edema, inflammation and necrosis. The main clinical manifestations include persistent or recurrent abdominal pain, weight loss and diabetes. Currently, the main methods for clinical treatment of CP include screening and treatment of complications, and there is a lack of specific therapies to delay or reverse the progression of CP.

[0003] The pathological characteristics of CP are that acinar cell damage leads to the infiltration of multiple inflammatory cells, mainly macrophages, which secrete a large number of proinflammatory cytokines, causing activation of pancreatic stellate cells (PSCs) in the pancreatic tissue microenvironment, and then produce a large amount of extracellular matrix (ECM), which eventually develops into pancreatic fibrosis. Therefore, pancreatic fibrosis is the main lesion in the process of CP. Finding targeted drugs to inhibit pancreatic fibrosis can effectively block the occurrence and development of CP. Studies have shown that macrophage polarization plays an important role in pancreatic fibrosis, and inhibiting macrophage M2 polarization can improve pancreatic inflammation and relieve pancreatic fibrosis.

[0004] Adenosylcobalamin is a homologue of vitamin B12, which maintains cell growth and proliferation as well as the intact structure of nerve myelin sheath. It is mainly used to treat various anemias such as megaloblastic, malnutrition and pregnancy, as well as inflammatory diseases such as multiple neuritis and radiculitis.

[0005] So far, there is no research report on the application of adenosylcobalamin in the prevention and treatment of CP. Therefore, actively exploring the role of adenosylcobalamin in macrophage M2 polarization and pancreatic fibrosis caused by CP has very important academic value and clinical significance. Summary of the invention

[0006] The purpose of the present invention is to provide a new application of adenosylcobalamin, to study the role of adenosylcobalamin in macrophage M2 polarization and pancreatic fibrosis caused by CP, and to provide a possible clinical medication scheme for the prevention and treatment of CP.

[0007] In order to achieve the above-mentioned invention object, the present invention adopts the following technical solutions:

[0008] The invention provides application of adenosylcobalamin in preparing a medicine for treating or preventing chronic pancreatitis.

[0009] The structural formula of adenosylcobalamin is as follows:

[0010]

[0011] In the above technical solution, further, the application is to inhibit macrophage M2 polarization.

[0012] In the above technical solution, further, the application is to reduce pancreatic fibrosis caused by chronic pancreatitis.

[0013] In the above technical solution, further, the application is to reduce the changes in pancreatic tissue caused by chronic pancreatitis.

[0014] The present invention also provides a pharmaceutical composition for preventing or treating chronic pancreatitis, wherein the pharmaceutical composition uses adenosylcobalamin as an active ingredient.

[0015] In the above technical solution, further, the drug is an oral preparation or an injectable preparation.

[0016] Compared with the prior art, the present invention has the following beneficial effects: the present invention provides the use of adenosylcobalamin in drugs for treating or preventing chronic pancreatitis, and experiments have verified that adenosylcobalamin can inhibit macrophage M2 polarization, reduce CP-induced pancreatic tissue atrophy, obvious nodules and other damage, and pathological changes such as pancreatic fibrosis. The present invention provides a basis for drug screening for chronic pancreatitis. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 Biochemical kits were used to detect the effects of adenosylcobalamin on the levels of CRE, BUN, ALT and AST in mouse serum. Data are presented as mean ± SEM. Cob, adenosylcobalamin; CRE, creatinine; BUN, urea nitrogen; ALT, alanine aminotransferase; AST, aspartate aminotransferase.

[0018] Figure 2 HE staining was used to observe the effect of adenosylcobalamin on the pathological changes of the heart, liver, spleen, lung, kidney, brain, small intestine and pancreas of mice (400×). Cob, adenosylcobalamin.

[0019] Figure 3 Flow cytometry results; (a) Flow cytometry detection of the effect of adenosylcobalamin on the content of M2 macrophages in mouse spleen tissue; (b) Statistical graph of mouse spleen flow cytometry results. Data are expressed as mean ± SEM; Compared with the CP group, ## P < 0.01. Cob, adenosylcobalamin.

[0020] Figure 4 F4-80-CD206 double immunofluorescence staining results; (a) F4-80 + -CD206 +Double immunofluorescence staining to observe the effect of adenosylcobalamin on M2 macrophages in mouse pancreatic tissue; (b) Mouse pancreatic F4-80 + -CD206 + Statistical graph of double immunofluorescence staining results. Data are expressed as mean ± SEM; compared with the Ctrl group, ** P<0.01; compared with the CP group, ## P < 0.01. Cob, adenosylcobalamin.

[0021] Figure 5 HE staining results; (a) HE staining to observe the effect of adenosylcobalamin on pancreatic tissue pathological damage in CP mice; (b) Statistical graph of HE staining results of mouse pancreas. Data are expressed as mean ± SEM; Compared with the Ctrl group, ** P<0.01; compared with the CP group, ## P < 0.01. Cob, adenosylcobalamin.

[0022] Figure 6 Masson staining results; (a) Masson staining to observe the effect of adenosylcobalamin on pancreatic fibrosis in CP mice; (b) Statistical graph of Masson staining results of mouse pancreas. Data are expressed as mean ± SEM; Compared with the Ctrl group, ** P<0.01; compared with the CP group, # P < 0.05. Cob, adenosylcobalamin. DETAILED DESCRIPTION

[0023] The present invention is further described below in conjunction with specific embodiments, but the present invention is not limited in any way.

[0024] Example 1

[0025] 1. Experimental plan:

[0026] 1) In vivo toxicity experiment: 16 C57BL / 6 mice were randomly divided into 2 groups (control group (Ctrl) and adenosylcobalamin group (Cob), n=8) after one week of adaptation to the environment to evaluate the in vivo toxicity of adenosylcobalamin. Mice in the Ctrl and Cob groups were intraperitoneally injected with normal saline and adenosylcobalamin (0.2 mg / kg / day), respectively. After 4 weeks, C57BL / 6 mice were anesthetized with isoflurane, and blood was collected from the abdominal aorta to collect serum and frozen in a -80°C refrigerator. Subsequently, biochemical kits were used to detect ALT, AST, BUN, CRE and other indicators in mouse serum; and pancreas, heart, liver, spleen, lung, kidney, brain, intestine and other tissues were fixed in 10% formaldehyde for HE staining to detect the in vivo toxicity of adenosylcobalamin.

[0027] 2) In vivo efficacy experiment: 24 C57BL / 6 mice were randomly divided into 3 groups (control group (Ctrl), chronic pancreatitis group (CP), CP+adenosylcobalamin group (CP+Cob), n=8) after one week of adaptation to the environment, so as to evaluate the in vivo efficacy of adenosylcobalamin. The CP mouse model was established by intraperitoneal injection of caerulein. C57BL / 6 mice were intraperitoneally injected with caerulein (50μg / kg) on ​​Mondays, Wednesdays, and Fridays every week, with an interval of 1h / time, 6 times a day, for 8 weeks; adenosylcobalamin was intraperitoneally injected 4 weeks after the start of CP modeling (0.2mg / kg / day, for 4 weeks); mice in the Ctrl group were intraperitoneally injected with an equal amount of normal saline at the same dosing frequency. After 8 weeks, C57BL / 6 mice were anesthetized with isoflurane, and blood was collected from the abdominal aorta to be killed; spleen tissue was obtained for trypsin digestion, and M2 macrophages (F480 + -CD206 + The pancreatic tissues were fixed in 10% formaldehyde for subsequent detection of M2 macrophage content and HE and Masson staining to evaluate the effect of adenosylcobalamin on CP-induced pancreatic injury and fibrotic lesions.

[0028] 2. Experimental results:

[0029] 1) Evaluation of the in vivo toxicity of adenosylcobalamin

[0030] like Figure 1 As shown in the figure, after 4 weeks of administration of adenosylcobalamin (0.2 mg / kg / day), the levels of CRE, BUN, ALT and AST in the mouse serum did not change significantly compared with the Ctrl group. HE staining results also showed that the heart, liver, spleen, lung, kidney, brain, small intestine and pancreas of the mice in the adenosylcobalamin group were normal in morphology, and there was no obvious pathological damage compared with the Ctrl group mice ( Figure 2 Therefore, intraperitoneal injection of adenosylcobalamin (0.2 mg / kg / day, 4 weeks) had no obvious toxic side effects on C57BL / 6 mice.

[0031] 2) Adenosylcobalamin improves pancreatic injury and fibrotic lesions in CP mice by reducing M2 polarization of macrophages

[0032] Flow cytometry was used to detect the content of M2 macrophages in spleen tissue. Figure 3 As shown in ab, compared with CP, the adenosylcobalamin group was able to significantly reduce F4-80 + -CD206 + The number of double positive cells, namely M2 macrophages. In addition, F4-80 + -CD206 +Double immunofluorescence results showed that compared with the Ctrl group, the expression of F4-80 (red fluorescence area) and CD206 (green fluorescence area) in the pancreatic tissue of CP mice was significantly upregulated, that is, the amount of M2 macrophages infiltrating in the pancreatic tissue was significantly increased, and the adenosylcobalamin administration group was able to significantly reduce the polarization of M2 macrophages ( Figure 4 ).

[0033] HE staining was used to evaluate the role of adenosylcobalamin in CP. Figure 5 As shown in the figure, compared with the Ctrl group, the pancreatic tissue of the mice in the CP group was atrophic, whitened, and had obvious nodules, while the pancreatic tissue damage of the mice in the adenosylcobalamin group was significantly alleviated. The results of Masson staining showed that the blue collagen fibrosis area in the pancreatic lesions of CP-induced mice increased, but the collagen fibrosis area in the adenosylcobalamin group was significantly reduced ( Figure 6 ). Therefore, adenosylcobalamin can significantly inhibit the M2 polarization of macrophages and reduce pancreatic damage and fibrosis caused by CP.

[0034] From the above results, it can be seen that adenosylcobalamin can inhibit the M2 polarization of macrophages, thereby reducing pancreatic damage and fibrosis in CP mice, and is a good candidate drug for the clinical prevention and treatment of CP.

Claims

1. Application of adenosylcobalamin in the preparation of drugs for treating or preventing chronic pancreatitis.

2. The use according to claim 1, characterized in that: The application is to inhibit macrophage M2 polarization.

3. The use according to claim 1, characterized in that: The application is to reduce pancreatic fibrosis caused by chronic pancreatitis.

4. The use according to claim 1, characterized in that: The application is to reduce the pancreatic tissue changes caused by chronic pancreatitis.

5. Use of a pharmaceutical composition in the preparation of a drug for preventing or treating chronic pancreatitis, wherein the pharmaceutical composition contains adenosylcobalamin as an active ingredient.

6. The use according to claim 5, characterized in that: The medicine is an oral preparation or an injection preparation.

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