CN108635571B - Application of irisin in preparation of medicine for preventing and treating severe acute pancreatitis - Google Patents
Application of irisin in preparation of medicine for preventing and treating severe acute pancreatitis Download PDFInfo
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- CN108635571B CN108635571B CN201810797454.4A CN201810797454A CN108635571B CN 108635571 B CN108635571 B CN 108635571B CN 201810797454 A CN201810797454 A CN 201810797454A CN 108635571 B CN108635571 B CN 108635571B
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- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
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Abstract
The invention belongs to the field of medicines, and particularly discloses application of irisin (irisin) in a medicine for preventing and treating severe acute pancreatitis, wherein in the severe acute pancreatitis of a mouse, exogenous irisin is injected intravenously, so that the pancreatic function can be obviously improved, the endoplasmic reticulum stress of pancreatic cells can be relieved, and the pancreatic tissue damage and cell death can be reduced, which shows that irisin can obviously improve the severe acute pancreatitis; the irisin is applied to medicine development, when acute pancreatitis is diagnosed in an emergency, the local inflammatory reaction is developed into systemic inflammatory response syndrome or the chain type expansion effect of the systemic inflammatory response syndrome is relieved when the acute pancreatitis is blocked and prevented, so that the intensive pancreatitis is inhibited, and local organs are protected and the multi-organ functional failure syndrome is prevented; the invention provides a brand new choice and idea for preventing and treating severe acute pancreatitis at present.
Description
Technical Field
The invention relates to the field of medicines, in particular to application of irisin in medicines for preventing and treating severe acute pancreatitis.
Background
Acute Pancreatitis (AP) is a common clinical Acute abdomen, wherein 20-25% of patients have clinically serious illness, local or systemic complications can occur, multiple organ failure occurs, and even life is threatened, and the Acute Pancreatitis (SAP) is called Severe Acute Pancreatitis. SAP has complex etiology and incomplete pathogenesis, and is estimated by the world health organization to become the first cause of death of pancreatic diseases in 2020. Through research in recent 30 years, the treatment effect of severe acute pancreatitis in China is remarkably improved, and the treatment view point reaches consensus in principle, namely, the individual treatment scheme for distinguishing different causes and different disease stages. At present, the overall survival rate of the treatment effect of acute pancreatitis reaches 83%, wherein the survival rate of operative treatment reaches more than 70%, and the survival rate of non-operative treatment reaches about 90%. Compared with the past, the curative effect is favorable, but is not satisfactory, because 20-30% of patients cannot be cured, the treatment course is long, the cost is high, and the pain of the patients is large. The most important reason for influencing the curative effect is that specific treatment which can really block the development of the disease state is lacked in the early stage of the disease course; the fulminant pancreatitis still has high mortality rate, and the survival rate of the fulminant pancreatitis is difficult to improve by only depending on operation or non-operation treatment; in different stages of the disease course, the selection of the surgical indication and the mastering of the surgical time, even in the aspects of the surgical method and the drainage technology, the difference in cognition often exists, so that the surgical time is delayed, and the disease focus is remained; the understanding of serious complications such as deep fungal infection and pancreatic encephalopathy is insufficient, and the mortality rate and the disability rate of the diseases are still high.
At present, the prevention of severe acute pancreatitis is mainly active surgical drainage, effective life support treatment is provided, and the progression of waterfall chain reaction of systemic inflammation is inhibited.
Irisin is a muscle factor found in 2012, and studies have shown that irisin can promote brown cell-like changes in white adipocytes of mice, and plays an important role in energy metabolism and type 2 diabetes. Irisin is a secretable polypeptide fragment formed by proteolysis of the fibronectin type III component comprising protein 5(fibronectin type IIIdomain-associating protein 5, FNDC 5). At present, no report on the application of irisin to severe acute pancreatitis exists.
Disclosure of Invention
The invention aims to provide application of irisin in preparing a medicine for preventing and treating severe acute pancreatitis.
In order to achieve the purpose, the invention provides the following technical scheme:
the invention provides application of irisin in preparing a medicine for preventing and treating severe acute pancreatitis.
The severe acute pancreatitis refers to a mouse model of the severe acute pancreatitis induced by L-arginine.
The irisin can relieve pancreatic dysfunction caused by severe acute pancreatitis.
The irisin can relieve active oxygen injury of pancreatic tissue caused by severe acute pancreatitis.
The irisin can relieve pancreatic cell mitochondria injury caused by severe acute pancreatitis.
The irisin can relieve the endoplasmic reticulum stress of pancreatic cells caused by severe acute pancreatitis.
The effective benefits of the invention are: the irisin is applied to medicine development, is injected into the vein when acute pancreatitis is diagnosed in emergency, and has the effects of blocking and preventing local inflammatory reaction from progressing to systemic inflammatory response syndrome or relieving chain enlargement of the systemic inflammatory response syndrome when pancreatitis is diagnosed, so that the severity of pancreatitis is inhibited, and local organs are protected and multi-organ failure syndrome is prevented. The invention provides a brand new choice and idea for preventing/treating the severe acute pancreatitis at present.
Drawings
FIG. 1: the irisin is injected into abdominal cavity to relieve pancreatic dysfunction caused by severe acute pancreatitis.
FIG. 2: irisin is injected into abdominal cavity to relieve pancreatic tissue injury caused by severe acute pancreatitis.
FIG. 3: the irisin is injected into the abdominal cavity to relieve the pancreatic cell mitochondria damage caused by severe acute pancreatitis.
FIG. 4: irisin is injected into abdominal cavity to relieve the endoplasmic reticulum stress of pancreatic cells caused by severe acute pancreatitis.
FIG. 5: the molecular structure of irisin.
Detailed Description
The invention is further described with reference to the drawings and the following detailed description, which are not intended to limit the invention in any way. The irisin used was synthesized by bio-corporation and commercially available. Reagents, methods and apparatus used in the present invention are conventional in the art unless otherwise indicated.
Example 1: 100 micrograms of irisin was dissolved in 10 ml of physiological saline and formulated into 10ug/ml injection.
The injection is taken as an example to research the effect of irisin on severe acute pancreatitis.
And (5) animal experiments.
Experimental animal one: 20-25g of male C57BL/6J mice were randomly divided into 4 groups of 6 mice:
normal control group (Sham);
control group of severe acute pancreatitis: the mouse is injected with 4g/kg (Vehicle) of 8 percent L-arginine in the abdominal cavity;
high-dose irisin treatment group for severe acute pancreatitis: mice were injected intraperitoneally with 4g/kg of 8% L-arginine and 2 hours later were given 250. mu.g/kg of Irisin (H-Irisin).
Low dose irisin treatment group for severe acute pancreatitis: mice were injected intraperitoneally with 4g/kg of 8% L-arginine and 2 hours later were given 50ug/kg of Irisin (L-Irisin).
II, secondly: preparing a severe acute pancreatitis model: vehicle mice were injected intraperitoneally with 4g/kg of 8% L-arginine to create an SAP model of severe acute pancreatitis. The Sham group was injected with saline of the same volume intraperitoneally; the mice in the H-Irisin group are given 250 mu g/kg of Irisin 2 hours after 4g/kg of 8 percent L-arginine is injected into the abdominal cavity. The L-Irisin mice are injected with 4g/kg of 8% L-arginine in the abdominal cavity for 2 hours and then are given 50ug/kg of Irisin.
Anesthetizing each mouse under isoflurane after injecting irisin for 24h to obtain a blood sample, and detecting the index of serum pancreatitis; after euthanasia of the mice, pancreatic tissues are taken, and part of pancreas is fixed by 4% paraformaldehyde for H & E staining; refrigerating the rest fresh pancreatic tissues at-80 ℃ for detecting indexes such as pancreatic oxidative stress indexes and mitochondrial damage.
Thirdly, the method comprises the following steps: experimental results;
1. and (3) the irisin improves the pancreatic function of the mice after the severe acute pancreatitis. The kit is used for detecting the contents of amylase and lipase in serum samples of each group of mice. The results are shown in FIG. 1, where Sham is the blank set without intervention; vihicle is a severe acute pancreatitis control group; H-Irisin is a high-dose Irisin treatment group for severe acute pancreatitis; L-Irisin is a low-dose Irisin treatment group for severe acute pancreatitis. The graph indicates the difference compared with the Sham group; # indicates a difference from the Vihicle group; the color is different from the H-irisin group.
The results show that: irisin can significantly improve the pancreatic function of mice after severe acute pancreatitis.
2. Irisin can be used for treating pancreatic tissue injury after severe acute pancreatitis of mouse. The results are shown in FIG. 2, and the fixed pancreatic tissue was embedded in paraffin blocks and sectioned and stained with hematoxylin and eosin (H & E). Results were scored histologically and necrotic area was measured. The tissue scoring method comprises the following steps: the method comprises the following 6 indexes: cytoplasm discoloration, vacuolation, nucleus shrinkage, nucleus fragmentation, nucleus regression and erythrocyte stasis, wherein each index is 1 (light), 2 (medium) and 3 (heavy), and the total score of the above 6 indexes is the average value of 6 mice. The necrotic area was estimated by H & E outcome measurements. The graph indicates the difference compared with the Sham group; # indicates a difference from the Vihicle group; the color is different from the H-irisin group.
The results show that: the caucasin can obviously relieve the pancreatic tissue injury of mice after severe acute pancreatitis.
3. Irisin improves the condition of mouse pancreatic tissue mitochondria damage after severe acute pancreatitis. Indexes such as PGC-1 alpha, DRP-1, ATPB, PINK and the like in pancreatic tissues of each group of mice are detected by using a Western-blot technology. The results are shown in FIG. 3, where Sham is the blank set without intervention; vihicle is a severe acute pancreatitis control group; H-Irisin is a high-dose Irisin treatment group for severe acute pancreatitis; L-Irisin is a low-dose Irisin treatment group for severe acute pancreatitis. The graph indicates the difference compared with the Sham group; # indicates a difference from the Vihicle group; the color is different from the H-irisin group.
The results show that: irisin can significantly reduce the injury of the mitochondria of pancreatic cells after severe acute pancreatitis of mice.
Therefore, the invention can be used for preparing the medicine for preventing/treating the severe acute pancreatitis.
4. Irisin improves the stress of pancreatic endoplasmic reticulum after severe acute pancreatitis of mice. And (3) detecting indexes such as IRE 1-alpha, GRP78 and the like in pancreatic tissues of each group of mice by using a Western-blot technology. The results are shown in FIG. 4, where Sham is the blank set without intervention; vihicle is a severe acute pancreatitis control group; H-Irisin is a high-dose Irisin treatment group for severe acute pancreatitis; L-Irisin is a low-dose Irisin treatment group for severe acute pancreatitis. The graph indicates the difference compared with the Sham group; # indicates a difference from the Vihicle group; the color is different from the H-irisin group.
The results show that: the irisin can obviously relieve the stress condition of the endoplasmic reticulum of pancreatic cells after the severe acute pancreatitis of the mice.
Data were all mean ± standard deviation. Statistical methods using SNK test, P <0.05 was statistically different.
Example (b): 100 micrograms of irisin was dissolved in 5 ml of physiological saline and formulated into 20ug/ml injection.
The injection is taken as an example to research the effect of irisin on severe acute pancreatitis.
And (5) animal experiments.
Experimental animal one: 20-25g of male C57BL/6J mice were randomly divided into 4 groups of 6 mice:
normal control group (Sham);
control group of severe acute pancreatitis: the mouse is injected with 4g/kg (Vehicle) of 8 percent L-arginine in the abdominal cavity;
high-dose irisin treatment group for severe acute pancreatitis: mice were injected intraperitoneally with 4g/kg of 8% L-arginine and 2 hours later were given 250. mu.g/kg of Irisin (H-Irisin).
Low dose irisin treatment group for severe acute pancreatitis: mice were injected intraperitoneally with 4g/kg of 8% L-arginine and 2 hours later were given 50ug/kg of Irisin (L-Irisin).
II, secondly: preparing a severe acute pancreatitis model: vehicle mice were injected intraperitoneally with 4g/kg of 8% L-arginine to create an SAP model of severe acute pancreatitis. The Sham group was injected with saline of the same volume intraperitoneally; the mice in the H-Irisin group are given 250 mu g/kg of Irisin 2 hours after 4g/kg of 8 percent L-arginine is injected into the abdominal cavity. The L-Irisin mice are injected with 4g/kg of 8% L-arginine in the abdominal cavity for 2 hours and then are given 50ug/kg of Irisin.
Anesthetizing each mouse under isoflurane after injecting irisin for 24h to obtain a blood sample, and detecting the index of serum pancreatitis; after euthanasia of the mice, pancreatic tissues are taken, and part of pancreas is fixed by 4% paraformaldehyde for H & E staining; refrigerating the rest fresh pancreatic tissues at-80 ℃ for detecting indexes such as pancreatic oxidative stress indexes and mitochondrial damage.
Thirdly, the method comprises the following steps: experimental results;
1. and (3) the irisin improves the pancreatic function of the mice after the severe acute pancreatitis. The kit is used for detecting the contents of amylase and lipase in serum samples of each group of mice. The results are shown in FIG. 1, where Sham is the blank set without intervention; vihicle is a severe acute pancreatitis control group; H-Irisin is a high-dose Irisin treatment group for severe acute pancreatitis; L-Irisin is a low-dose Irisin treatment group for severe acute pancreatitis. The graph indicates the difference compared with the Sham group; # indicates a difference from the Vihicle group; the color is different from the H-irisin group.
The results show that: irisin can significantly improve the pancreatic function of mice after severe acute pancreatitis.
2. Irisin can be used for treating pancreatic tissue injury after severe acute pancreatitis of mouse. The results are shown in FIG. 2, and the fixed pancreatic tissue was embedded in paraffin blocks and sectioned and stained with hematoxylin and eosin (H & E). Results were scored histologically and necrotic area was measured. The tissue scoring method comprises the following steps: the method comprises the following 6 indexes: cytoplasm discoloration, vacuolation, nucleus shrinkage, nucleus fragmentation, nucleus regression and erythrocyte stasis, wherein each index is 1 (light), 2 (medium) and 3 (heavy), and the total score of the above 6 indexes is the average value of 6 mice. The necrotic area was estimated by H & E outcome measurements. The graph indicates the difference compared with the Sham group; # indicates a difference from the Vihicle group; the color is different from the H-irisin group.
The results show that: the caucasin can obviously relieve the pancreatic tissue injury of mice after severe acute pancreatitis.
3. Irisin improves the condition of mouse pancreatic tissue mitochondria damage after severe acute pancreatitis. Indexes such as PGC-1 alpha, DRP-1, ATPB, PINK and the like in pancreatic tissues of each group of mice are detected by using a Western-blot technology. The results are shown in FIG. 3, where Sham is the blank set without intervention; vihicle is a severe acute pancreatitis control group; H-Irisin is a high-dose Irisin treatment group for severe acute pancreatitis; L-Irisin is a low-dose Irisin treatment group for severe acute pancreatitis. The graph indicates the difference compared with the Sham group; # indicates a difference from the Vihicle group; the color is different from the H-irisin group.
The results show that: irisin can significantly reduce the injury of the mitochondria of pancreatic cells after severe acute pancreatitis of mice.
Therefore, the invention can be used for preparing the medicine for preventing/treating the severe acute pancreatitis.
4. Irisin improves the stress of pancreatic endoplasmic reticulum after severe acute pancreatitis of mice. And (3) detecting indexes such as IRE 1-alpha, GRP78 and the like in pancreatic tissues of each group of mice by using a Western-blot technology. The results are shown in FIG. 4, where Sham is the blank set without intervention; vihicle is a severe acute pancreatitis control group; H-Irisin is a high-dose Irisin treatment group for severe acute pancreatitis; L-Irisin is a low-dose Irisin treatment group for severe acute pancreatitis. The graph indicates the difference compared with the Sham group; # indicates a difference from the Vihicle group; the color is different from the H-irisin group.
The results show that: the irisin can obviously relieve the stress condition of the endoplasmic reticulum of pancreatic cells after the severe acute pancreatitis of the mice.
Data were all mean ± standard deviation. Statistical methods using SNK test, P <0.05 was statistically different.
Example 1: 100 micrograms of irisin was dissolved in 10 ml of physiological saline and formulated into 10ug/ml injection.
The injection is taken as an example to research the effect of irisin on severe acute pancreatitis.
And (5) animal experiments.
Experimental animal one: 20-25g of male C57BL/6J mice were randomly divided into 4 groups of 6 mice:
normal control group (Sham);
control group of severe acute pancreatitis: the mouse is injected with 4g/kg (Vehicle) of 8 percent L-arginine in the abdominal cavity;
high-dose irisin treatment group for severe acute pancreatitis: mice were injected intraperitoneally with 4g/kg of 8% L-arginine and 2 hours later were given 250. mu.g/kg of Irisin (H-Irisin).
Low dose irisin treatment group for severe acute pancreatitis: mice were injected intraperitoneally with 4g/kg of 8% L-arginine and 2 hours later were given 50ug/kg of Irisin (L-Irisin).
II, secondly: preparing a severe acute pancreatitis model: vehicle mice were injected intraperitoneally with 4g/kg of 8% L-arginine to create an SAP model of severe acute pancreatitis. The Sham group was injected with saline of the same volume intraperitoneally; the mice in the H-Irisin group are given 250 mu g/kg of Irisin 2 hours after 4g/kg of 8 percent L-arginine is injected into the abdominal cavity. The L-Irisin mice are injected with 4g/kg of 8% L-arginine in the abdominal cavity for 2 hours and then are given 50ug/kg of Irisin.
Anesthetizing each mouse under isoflurane after injecting irisin for 24h to obtain a blood sample, and detecting the index of serum pancreatitis; after euthanasia of the mice, pancreatic tissues are taken, and part of pancreas is fixed by 4% paraformaldehyde for H & E staining; refrigerating the rest fresh pancreatic tissues at-80 ℃ for detecting indexes such as pancreatic oxidative stress indexes and mitochondrial damage.
Thirdly, the method comprises the following steps: experimental results;
1. and (3) the irisin improves the pancreatic function of the mice after the severe acute pancreatitis. The kit is used for detecting the contents of amylase and lipase in serum samples of each group of mice. The results are shown in FIG. 1, where Sham is the blank set without intervention; vihicle is a severe acute pancreatitis control group; H-Irisin is a high-dose Irisin treatment group for severe acute pancreatitis; L-Irisin is a low-dose Irisin treatment group for severe acute pancreatitis. The graph indicates the difference compared with the Sham group; # indicates a difference from the Vihicle group; the color is different from the H-irisin group.
The results show that: irisin can significantly improve the pancreatic function of mice after severe acute pancreatitis.
2. Irisin can be used for treating pancreatic tissue injury after severe acute pancreatitis of mouse. The results are shown in FIG. 2, and the fixed pancreatic tissue was embedded in paraffin blocks and sectioned and stained with hematoxylin and eosin (H & E). Results were scored histologically and necrotic area was measured. The tissue scoring method comprises the following steps: the method comprises the following 6 indexes: cytoplasm discoloration, vacuolation, nucleus shrinkage, nucleus fragmentation, nucleus regression and erythrocyte stasis, wherein each index is 1 (light), 2 (medium) and 3 (heavy), and the total score of the above 6 indexes is the average value of 6 mice. The necrotic area was estimated by H & E outcome measurements. The graph indicates the difference compared with the Sham group; # indicates a difference from the Vihicle group; the color is different from the H-irisin group.
The results show that: the caucasin can obviously relieve the pancreatic tissue injury of mice after severe acute pancreatitis.
3. Irisin improves the condition of mouse pancreatic tissue mitochondria damage after severe acute pancreatitis. Indexes such as PGC-1 alpha, DRP-1, ATPB, PINK and the like in pancreatic tissues of each group of mice are detected by using a Western-blot technology. The results are shown in FIG. 3, where Sham is the blank set without intervention; vihicle is a severe acute pancreatitis control group; H-Irisin is a high-dose Irisin treatment group for severe acute pancreatitis; L-Irisin is a low-dose Irisin treatment group for severe acute pancreatitis. The graph indicates the difference compared with the Sham group; # indicates a difference from the Vihicle group; the color is different from the H-irisin group.
The results show that: irisin can significantly reduce the injury of the mitochondria of pancreatic cells after severe acute pancreatitis of mice.
Therefore, the invention can be used for preparing the medicine for preventing/treating the severe acute pancreatitis.
4. Irisin improves the stress of pancreatic endoplasmic reticulum after severe acute pancreatitis of mice. And (3) detecting indexes such as IRE 1-alpha, GRP78 and the like in pancreatic tissues of each group of mice by using a Western-blot technology. The results are shown in FIG. 4, where Sham is the blank set without intervention; vihicle is a severe acute pancreatitis control group; H-Irisin is a high-dose Irisin treatment group for severe acute pancreatitis; L-Irisin is a low-dose Irisin treatment group for severe acute pancreatitis. The graph indicates the difference compared with the Sham group; # indicates a difference from the Vihicle group; the color is different from the H-irisin group.
The results show that: the irisin can obviously relieve the stress condition of the endoplasmic reticulum of pancreatic cells after the severe acute pancreatitis of the mice.
Data were all mean ± standard deviation. Statistical methods using SNK test, P <0.05 was statistically different.
Claims (6)
1. Application of irisin in preparing medicine for treating severe acute pancreatitis is provided.
2. The use of claim 1, wherein the severe acute pancreatitis is a mouse model of L-arginine-induced severe acute pancreatitis.
3. The use according to claim 1, wherein irisin is capable of alleviating pancreatic dysfunction associated with severe acute pancreatitis.
4. The use according to claim 1, wherein said irisin reduces pancreatic tissue reactive oxygen species damage caused by severe acute pancreatitis.
5. The use according to claim 1, wherein said irisin reduces pancreatic cell mitochondrial damage resulting from severe acute pancreatitis.
6. The use according to claim 1, wherein irisin reduces pancreatic endoplasmic reticulum stress caused by severe acute pancreatitis.
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Effective date of registration: 20220225 Address after: 518066 1201, building a, phase I, Qianhai economic and Trade Center, China Merchants Group, No. 151, Zimao West Street, Nanshan street, Qianhai Shenzhen Hong Kong cooperation zone, Shenzhen, Guangdong Province Patentee after: Chonghao Technology Co.,Ltd. Address before: 710061 No. 277, Yanta West Road, Shaanxi, Xi'an Patentee before: THE FIRST AFFILIATED HOSPITAL OF MEDICAL COLLEGE OF XI'AN JIAOTONG University |