CN108635571A - Irisin(irisin)Application in preventing and treating Severe Acute Pancreatitis SAP drug - Google Patents

Irisin(irisin)Application in preventing and treating Severe Acute Pancreatitis SAP drug Download PDF

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Publication number
CN108635571A
CN108635571A CN201810797454.4A CN201810797454A CN108635571A CN 108635571 A CN108635571 A CN 108635571A CN 201810797454 A CN201810797454 A CN 201810797454A CN 108635571 A CN108635571 A CN 108635571A
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irisin
acute pancreatitis
severe acute
pancreatitis sap
sap
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CN108635571B (en
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吴荣谦
任凡
任一凡
吕毅
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Chonghao Technology Co.,Ltd.
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First Affiliated Hospital of Medical College of Xian Jiaotong University
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    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00Medicinal preparations containing peptides
    • A61K38/16Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/39Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin, cold insoluble globulin [CIG]
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/18Drugs for disorders of the alimentary tract or the digestive system for pancreatic disorders, e.g. pancreatic enzymes

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Abstract

The invention belongs to field of medicaments, specifically disclose application of the irisin (irisin) in preventing and treating Severe Acute Pancreatitis SAP drug, in mouse Severe Acute Pancreatitis SAP, pancreas function can be significantly improved by being injected intravenously exogenous irisin, mitigate pancreatic cell er stress, pancreatic tissue damage and cell death are reduced, shows that irisin can significantly improve Severe Acute Pancreatitis SAP;Irisin is applied to drug development; it is injected intravenously when acute pancreatitis is made a definite diagnosis in emergency treatment; when blocking and preventing pancreatitis; local inflammation reaction progresses to systemic inflammatory response syndrome or mitigates the widened effect of systemic inflammatory response syndrome chain type; to inhibit pancreatitis worsening, plays the local internal organs of protection and prevent the generation of multiple organ failure syndrome;The present invention provides completely new selection and thinking to prevent at present and treating Severe Acute Pancreatitis SAP.

Description

Application of the irisin (irisin) in preventing and treating Severe Acute Pancreatitis SAP drug
Technical field
The present invention relates to field of medicaments, and in particular to irisin (irisin) is preventing and treating Severe Acute Pancreatitis SAP medicine Application in object.
Background technology
Acute pancreatitis (Acute Pancreatitis, AP) is clinical common acute abdomen, the wherein disease of 20%-25% People's clinically state of an illness weight, may occur in which locally or systemically complication, multiple organ failure or even threat to life occurs, referred to as heavy Disease acute pancreatitis (Severe Acute Pancreatitis, SAP).The SAP causes of disease are complicated, and pathogenesis is not yet completely clear, Estimate according to the World Health Organization, will become the No.1 cause of death of pancreatic disease to the year two thousand twenty SAP.By nearly research in 30 years, I The therapeutic effect of state's Severe Acute Pancreatitis SAP has had significant raising, treatment viewpoint also to reach common recognition in principle, that Just be to discriminate between different pathogeny, different stadium individualized treatment scheme.The therapeutic effect of acute pancreatitis at present, overall survival Up to 83%, the wherein survival rate of operative treatment is up to 70% or more, and non-operative treatment person is up to 90% or so.Compared with the past, this One curative effect is gratifying, but can't be satisfactory, because also 20%~30% patient cannot cure, and the course for the treatment of is long, Costly, patient's pain is big.Affect the treatment it is most important the reason is that, also lacked early stage the course of disease really can blocking condition development Specific treatment;Operation treatment still has the very high death rate, simple to be difficult to improve by operation or non-operative treatment Its survival rate;In the different phase of the course of disease, the selection of surgical indication and the grasp of opportunity of operation, or even in operation method, drainage Technical aspect usually there is also the difference in understanding, makes opportunity of operation be delayed, Residue;To severe complication such as fungi deep Infection and pancreatic encephalopathy understanding are insufficient, and the death rate and disability rate are still very high.
The prevention of Severe Acute Pancreatitis SAP is mainly positive operative drainage at present, gives effective life supportive treatment, suppression Systemic inflammatorome waterfall type chain reaction progress processed aggravates.
Irisin is a kind of muscle factor for being found in 2012, current study show that irisin can promote the white of mouse Adipocyte occurs brown cell sample and changes, and plays a significant role in energetic supersession and diabetes B.Irisin is type III Fibronectin component includes albumen 5 (fibronectin type IIIdomain-containing protein 5, FNDC5) What is formed after proteolysis can secrete polypeptide segment.Currently, any report that there is no irisin to be applied in Severe Acute Pancreatitis SAP Road.
Invention content
The purpose of the present invention is to provide irisins (irisin) in preventing and treating Severe Acute Pancreatitis SAP drug Using.
To achieve the goals above, the present invention provides following technical scheme:
The present invention provides application of the irisin (irisin) in preventing and treating Severe Acute Pancreatitis SAP drug.
The Severe Acute Pancreatitis SAP refers to the Severe Acute Pancreatitis SAP mouse model of L-arginine induction.
It is abnormal that the irisin can alleviate pancreas function caused by Severe Acute Pancreatitis SAP.
The irisin can mitigate pancreatic tissue Damage Induced by Reactive Oxygen Species caused by Severe Acute Pancreatitis SAP.
The irisin can mitigate pancreatic cell injury of mitochondria caused by Severe Acute Pancreatitis SAP.
The irisin can mitigate pancreatic cell er stress caused by Severe Acute Pancreatitis SAP.
The present invention effective income be:Irisin is applied to drug development, the vein when acute pancreatitis is made a definite diagnosis in emergency treatment Injection blocks and prevents when pancreatitis that local inflammation reaction progresses to systemic inflammatory response syndrome or to mitigate systemic inflammatorome anti- The widened effect of syndrome chain type is answered, to inhibit pancreatitis worsening, the local internal organs of protection is played and prevents multiple organ dysfunction The generation of wasting syndrome.The present invention provides completely new selection and thinking for current preventing/treating Severe Acute Pancreatitis SAP.
Description of the drawings
Fig. 1:It is abnormal that pancreas function caused by Severe Acute Pancreatitis SAP is alleviated in irisin intraperitoneal injection.
Fig. 2:Irisin intraperitoneal injection mitigates pancreatic tissue caused by Severe Acute Pancreatitis SAP and damages.
Fig. 3:Irisin intraperitoneal injection mitigates pancreatic cell injury of mitochondria caused by Severe Acute Pancreatitis SAP.
Fig. 4:Irisin intraperitoneal injection mitigates pancreatic cell er stress caused by Severe Acute Pancreatitis SAP.
Fig. 5:The molecular structure of irisin.
Specific implementation mode
It is further illustrated the present invention below in conjunction with Figure of description and specific embodiment, but embodiment is not to the present invention It limits in any form.Irisin used is synthesized by biotech firm, can directly buy commercially available product.Unless stated otherwise, originally The reagent used, method and apparatus are invented as the art conventional reagent, method and apparatus.
Embodiment 1:By 100 microgram irisins, 10 milliliters of physiological saline solutions, it is configured to 10ug/ml injections.
Effect of the irisin to Severe Acute Pancreatitis SAP is studied by taking this injection as an example.
Zoopery.
One, experimental animals:20-25g male C 57 BL/6 J mouses are taken to be randomly divided into 4 groups, every group 6:
Normal group (Sham);
Severe Acute Pancreatitis SAP control group:Mouse peritoneal injects 8%L- arginine 4g/kg (Vehicle);
Severe Acute Pancreatitis SAP high dose irisin treatment group:Mouse peritoneal injects 8%L- arginine 4g/kg, after 2 hours Give 250 μ g/kg irisins (H-Irisin).
Severe Acute Pancreatitis SAP low dosage irisin treatment group:Mouse peritoneal injects 8%L- arginine 4g/kg, after 2 hours Give 50ug/kg irisins (L-Irisin).
Two:Model of Severe Acute Pancreatitis makes:Vehicle mouse make severe through 8%L- arginine 4g/kg is injected intraperitoneally Acute pancreatitis SAP models.Sham groups are through being injected intraperitoneally equal volume physiological saline;H-Irisin groups mouse is through intraperitoneal injection Give within 2 hours after 8%L- arginine 4g/kg 250 μ g/kg Irisin.L-Irisin group mouse peritoneals inject 8%L- arginine 4g/kg gives 50ug/kg irisins after 2 hours.
Anesthesia takes blood specimen to above-mentioned each mouse under isoflurane after irisin is injected for 24 hours, detects serum pancreatitis index;It is small Pancreatic tissue is taken after mouse euthanasia, part pancreas is taken to fix row H&E dyeing with 4% paraformaldehyde;Remaining fresh pancreatic tissue -80 Degree refrigeration, to detect the indexs such as pancreas oxidative stress index and injury of mitochondria.
Three:Experimental result;
1. irisin improves situation to pancreas function after mouse Severe Acute Pancreatitis SAP.Each group mouse blood is detected with kit Amylase, lipase content in clear sample.The result is shown in Figure 1, Sham is the blank group that do not intervene in figure;Vihicle is severe Acute pancreatitis control group;H-Irisin is Severe Acute Pancreatitis SAP high dose Irisin treatment groups;L-Irisin is that severe is anxious Property pancreatitis low dosage Irisin treatment groups.* is indicated in figure and Sham groups are more variant;# is indicated and Vihicle groups have difference It is different;※ is indicated and H-irisin groups are variant.
The result shows that:Irisin can significantly improve pancreas function situation after mouse Severe Acute Pancreatitis SAP.
2. irisin is to pancreatic tissue degree of impairment after mouse Severe Acute Pancreatitis SAP.As a result see Fig. 2, fixed pancreas group It knits and is embedded and be sliced with paraffin wax block, with haematoxylin eosin stains (H&E).Using histological score and downright bad face is measured to result Product.Organize methods of marking:Including following 6 indexs:Cytoplasm colour fading, vacuolation, karyopycnosis, karyorrhexis, core are subsided and red thin Born of the same parents are retarded by silt, and each index is divided into 1 (light), 2 (in), 3 (weight)s, obatained score is that 6 mouse that must be divided into of above 6 indexs are put down Mean value.Necrosis area is estimated by H&E outcome measurements.* is indicated in figure and Sham groups are more variant;# is indicated and Vihicle groups It is variant;※ is indicated and H-irisin groups are variant.
The result shows that:Pancreatic tissue is damaged after tail element can significantly mitigate mouse Severe Acute Pancreatitis SAP.
3. irisin improves situation to pancreatic tissue injury of mitochondria after mouse Severe Acute Pancreatitis SAP.Use Western- Blot technologies detect the indexs such as PGC-1 α, DRP-1, ATPB and PINK in each group mice pancreatic tissue.As a result Fig. 3 is seen, in figure Sham is the blank group that do not intervene;Vihicle is Severe Acute Pancreatitis SAP control group;H-Irisin is Severe Acute Pancreatitis SAP High dose Irisin treatment groups;L-Irisin is Severe Acute Pancreatitis SAP low dosage Irisin treatment groups.* expressions and Sham in figure Group is more variant;# is indicated and Vihicle groups are variant;※ is indicated and H-irisin groups are variant.
The result shows that:Irisin can significantly mitigate pancreatic cell injury of mitochondria after mouse Severe Acute Pancreatitis SAP.
Therefore, the present invention may be used as the preparation of preventing/treating Severe Acute Pancreatitis SAP drug.
4. irisin improves situation to pancreatic cell er stress after mouse Severe Acute Pancreatitis SAP.Use Western- Blot technologies detect the indexs such as IRE1- α and GRP78 in each group mice pancreatic tissue.As a result see Fig. 4, Sham is not have in figure The blank group of intervention;Vihicle is Severe Acute Pancreatitis SAP control group;H-Irisin is Severe Acute Pancreatitis SAP high dose Irisin treatment groups;L-Irisin is Severe Acute Pancreatitis SAP low dosage Irisin treatment groups.* is indicated in figure and Sham groups compare It is variant;# is indicated and Vihicle groups are variant;※ is indicated and H-irisin groups are variant.
The result shows that:Irisin can significantly mitigate pancreatic cell er stress feelings after mouse Severe Acute Pancreatitis SAP Condition.
Data are all made of means standard deviation.Statistical method is examined using SNK, P<0.05 has significant difference.
Embodiment:By 100 microgram irisins, 5 milliliters of physiological saline solutions, it is configured to 20ug/ml injections.
Effect of the irisin to Severe Acute Pancreatitis SAP is studied by taking this injection as an example.
Zoopery.
One, experimental animals:20-25g male C 57 BL/6 J mouses are taken to be randomly divided into 4 groups, every group 6:
Normal group (Sham);
Severe Acute Pancreatitis SAP control group:Mouse peritoneal injects 8%L- arginine 4g/kg (Vehicle);
Severe Acute Pancreatitis SAP high dose irisin treatment group:Mouse peritoneal injects 8%L- arginine 4g/kg, after 2 hours Give 250 μ g/kg irisins (H-Irisin).
Severe Acute Pancreatitis SAP low dosage irisin treatment group:Mouse peritoneal injects 8%L- arginine 4g/kg, after 2 hours Give 50ug/kg irisins (L-Irisin).
Two:Model of Severe Acute Pancreatitis makes:Vehicle mouse make severe through 8%L- arginine 4g/kg is injected intraperitoneally Acute pancreatitis SAP models.Sham groups are through being injected intraperitoneally equal volume physiological saline;H-Irisin groups mouse is through intraperitoneal injection Give within 2 hours after 8%L- arginine 4g/kg 250 μ g/kg Irisin.L-Irisin group mouse peritoneals inject 8%L- arginine 4g/kg gives 50ug/kg irisins after 2 hours.
Anesthesia takes blood specimen to above-mentioned each mouse under isoflurane after irisin is injected for 24 hours, detects serum pancreatitis index;It is small Pancreatic tissue is taken after mouse euthanasia, part pancreas is taken to fix row H&E dyeing with 4% paraformaldehyde;Remaining fresh pancreatic tissue -80 Degree refrigeration, to detect the indexs such as pancreas oxidative stress index and injury of mitochondria.
Three:Experimental result;
1. irisin improves situation to pancreas function after mouse Severe Acute Pancreatitis SAP.Each group mouse blood is detected with kit Amylase, lipase content in clear sample.The result is shown in Figure 1, Sham is the blank group that do not intervene in figure;Vihicle is severe Acute pancreatitis control group;H-Irisin is Severe Acute Pancreatitis SAP high dose Irisin treatment groups;L-Irisin is that severe is anxious Property pancreatitis low dosage Irisin treatment groups.* is indicated in figure and Sham groups are more variant;# is indicated and Vihicle groups have difference It is different;※ is indicated and H-irisin groups are variant.
The result shows that:Irisin can significantly improve pancreas function situation after mouse Severe Acute Pancreatitis SAP.
2. irisin is to pancreatic tissue degree of impairment after mouse Severe Acute Pancreatitis SAP.As a result see Fig. 2, fixed pancreas group It knits and is embedded and be sliced with paraffin wax block, with haematoxylin eosin stains (H&E).Using histological score and downright bad face is measured to result Product.Organize methods of marking:Including following 6 indexs:Cytoplasm colour fading, vacuolation, karyopycnosis, karyorrhexis, core are subsided and red thin Born of the same parents are retarded by silt, and each index is divided into 1 (light), 2 (in), 3 (weight)s, obatained score is that 6 mouse that must be divided into of above 6 indexs are put down Mean value.Necrosis area is estimated by H&E outcome measurements.* is indicated in figure and Sham groups are more variant;# is indicated and Vihicle groups It is variant;※ is indicated and H-irisin groups are variant.
The result shows that:Pancreatic tissue is damaged after tail element can significantly mitigate mouse Severe Acute Pancreatitis SAP.
3. irisin improves situation to pancreatic tissue injury of mitochondria after mouse Severe Acute Pancreatitis SAP.Use Western- Blot technologies detect the indexs such as PGC-1 α, DRP-1, ATPB and PINK in each group mice pancreatic tissue.As a result Fig. 3 is seen, in figure Sham is the blank group that do not intervene;Vihicle is Severe Acute Pancreatitis SAP control group;H-Irisin is Severe Acute Pancreatitis SAP High dose Irisin treatment groups;L-Irisin is Severe Acute Pancreatitis SAP low dosage Irisin treatment groups.* expressions and Sham in figure Group is more variant;# is indicated and Vihicle groups are variant;※ is indicated and H-irisin groups are variant.
The result shows that:Irisin can significantly mitigate pancreatic cell injury of mitochondria after mouse Severe Acute Pancreatitis SAP.
Therefore, the present invention may be used as the preparation of preventing/treating Severe Acute Pancreatitis SAP drug.
4. irisin improves situation to pancreatic cell er stress after mouse Severe Acute Pancreatitis SAP.Use Western- Blot technologies detect the indexs such as IRE1- α and GRP78 in each group mice pancreatic tissue.As a result see Fig. 4, Sham is not have in figure The blank group of intervention;Vihicle is Severe Acute Pancreatitis SAP control group;H-Irisin is Severe Acute Pancreatitis SAP high dose Irisin treatment groups;L-Irisin is Severe Acute Pancreatitis SAP low dosage Irisin treatment groups.* is indicated in figure and Sham groups compare It is variant;# is indicated and Vihicle groups are variant;※ is indicated and H-irisin groups are variant.
The result shows that:Irisin can significantly mitigate pancreatic cell er stress feelings after mouse Severe Acute Pancreatitis SAP Condition.
Data are all made of means standard deviation.Statistical method is examined using SNK, P<0.05 has significant difference.
Embodiment 1:By 100 microgram irisins, 10 milliliters of physiological saline solutions, it is configured to 10ug/ml injections.
Effect of the irisin to Severe Acute Pancreatitis SAP is studied by taking this injection as an example.
Zoopery.
One, experimental animals:20-25g male C 57 BL/6 J mouses are taken to be randomly divided into 4 groups, every group 6:
Normal group (Sham);
Severe Acute Pancreatitis SAP control group:Mouse peritoneal injects 8%L- arginine 4g/kg (Vehicle);
Severe Acute Pancreatitis SAP high dose irisin treatment group:Mouse peritoneal injects 8%L- arginine 4g/kg, after 2 hours Give 250 μ g/kg irisins (H-Irisin).
Severe Acute Pancreatitis SAP low dosage irisin treatment group:Mouse peritoneal injects 8%L- arginine 4g/kg, after 2 hours Give 50ug/kg irisins (L-Irisin).
Two:Model of Severe Acute Pancreatitis makes:Vehicle mouse make severe through 8%L- arginine 4g/kg is injected intraperitoneally Acute pancreatitis SAP models.Sham groups are through being injected intraperitoneally equal volume physiological saline;H-Irisin groups mouse is through intraperitoneal injection Give within 2 hours after 8%L- arginine 4g/kg 250 μ g/kg Irisin.L-Irisin group mouse peritoneals inject 8%L- arginine 4g/kg gives 50ug/kg irisins after 2 hours.
Anesthesia takes blood specimen to above-mentioned each mouse under isoflurane after irisin is injected for 24 hours, detects serum pancreatitis index;It is small Pancreatic tissue is taken after mouse euthanasia, part pancreas is taken to fix row H&E dyeing with 4% paraformaldehyde;Remaining fresh pancreatic tissue -80 Degree refrigeration, to detect the indexs such as pancreas oxidative stress index and injury of mitochondria.
Three:Experimental result;
1. irisin improves situation to pancreas function after mouse Severe Acute Pancreatitis SAP.Each group mouse blood is detected with kit Amylase, lipase content in clear sample.The result is shown in Figure 1, Sham is the blank group that do not intervene in figure;Vihicle is severe Acute pancreatitis control group;H-Irisin is Severe Acute Pancreatitis SAP high dose Irisin treatment groups;L-Irisin is that severe is anxious Property pancreatitis low dosage Irisin treatment groups.* is indicated in figure and Sham groups are more variant;# is indicated and Vihicle groups have difference It is different;※ is indicated and H-irisin groups are variant.
The result shows that:Irisin can significantly improve pancreas function situation after mouse Severe Acute Pancreatitis SAP.
2. irisin is to pancreatic tissue degree of impairment after mouse Severe Acute Pancreatitis SAP.As a result see Fig. 2, fixed pancreas group It knits and is embedded and be sliced with paraffin wax block, with haematoxylin eosin stains (H&E).Using histological score and downright bad face is measured to result Product.Organize methods of marking:Including following 6 indexs:Cytoplasm colour fading, vacuolation, karyopycnosis, karyorrhexis, core are subsided and red thin Born of the same parents are retarded by silt, and each index is divided into 1 (light), 2 (in), 3 (weight)s, obatained score is that 6 mouse that must be divided into of above 6 indexs are put down Mean value.Necrosis area is estimated by H&E outcome measurements.* is indicated in figure and Sham groups are more variant;# is indicated and Vihicle groups It is variant;※ is indicated and H-irisin groups are variant.
The result shows that:Pancreatic tissue is damaged after tail element can significantly mitigate mouse Severe Acute Pancreatitis SAP.
3. irisin improves situation to pancreatic tissue injury of mitochondria after mouse Severe Acute Pancreatitis SAP.Use Western- Blot technologies detect the indexs such as PGC-1 α, DRP-1, ATPB and PINK in each group mice pancreatic tissue.As a result Fig. 3 is seen, in figure Sham is the blank group that do not intervene;Vihicle is Severe Acute Pancreatitis SAP control group;H-Irisin is Severe Acute Pancreatitis SAP High dose Irisin treatment groups;L-Irisin is Severe Acute Pancreatitis SAP low dosage Irisin treatment groups.* expressions and Sham in figure Group is more variant;# is indicated and Vihicle groups are variant;※ is indicated and H-irisin groups are variant.
The result shows that:Irisin can significantly mitigate pancreatic cell injury of mitochondria after mouse Severe Acute Pancreatitis SAP.
Therefore, the present invention may be used as the preparation of preventing/treating Severe Acute Pancreatitis SAP drug.
4. irisin improves situation to pancreatic cell er stress after mouse Severe Acute Pancreatitis SAP.Use Western- Blot technologies detect the indexs such as IRE1- α and GRP78 in each group mice pancreatic tissue.As a result see Fig. 4, Sham is not have in figure The blank group of intervention;Vihicle is Severe Acute Pancreatitis SAP control group;H-Irisin is Severe Acute Pancreatitis SAP high dose Irisin treatment groups;L-Irisin is Severe Acute Pancreatitis SAP low dosage Irisin treatment groups.* is indicated in figure and Sham groups compare It is variant;# is indicated and Vihicle groups are variant;※ is indicated and H-irisin groups are variant.
The result shows that:Irisin can significantly mitigate pancreatic cell er stress feelings after mouse Severe Acute Pancreatitis SAP Condition.
Data are all made of means standard deviation.Statistical method is examined using SNK, P<0.05 has significant difference.

Claims (6)

1. application of the irisin (irisin) in preventing and treating Severe Acute Pancreatitis SAP drug.
2. application according to claim 1, which is characterized in that the Severe Acute Pancreatitis SAP refers to L-arginine induction Severe Acute Pancreatitis SAP mouse model.
3. application according to claim 1, which is characterized in that the irisin can alleviate Severe Acute Pancreatitis SAP and draw The pancreas function risen is abnormal.
4. application according to claim 1, which is characterized in that the irisin can mitigate Severe Acute Pancreatitis SAP and draw The pancreatic tissue Damage Induced by Reactive Oxygen Species risen.
5. application according to claim 1, which is characterized in that the irisin can mitigate Severe Acute Pancreatitis SAP and draw The pancreatic cell injury of mitochondria risen.
6. application according to claim 1, which is characterized in that the irisin can mitigate Severe Acute Pancreatitis SAP and draw The pancreatic cell er stress risen.
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