Rainfrog extract induces chronic pancreatitis animal model in mice

Rainfrog extract induces chronic pancreatitis animal model in mice

Rainfrog extract induces chronic pancreatitis animal model in mice：

1、 Experimental animal selection

• ‌Type and GenderC57BL/6 male mice (8-10 weeks old, weight 20-25g), some studies have used PRSS1 transgenic mice to enhance human chronic pancreatitis (CP) similarity
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• ‌Feeding conditionsSPF level environment, fasting for 12 hours before the experiment (free drinking water)
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2、 Preparation and administration plan of rain frog extract

1. ‌Preparation of mother liquor‌

• Dissolve 1 mg of rain frog extract in 1 mL of physiological saline to prepare a 1 mg/mL mother liquor, filter and sterilize, and then pack and freeze for storage
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2. ‌Concentration of working fluid‌

• Dilute to 5-10 μ g/mL (physiological saline) before use, with a commonly used injection dose of 50 μ g/kg.

3. ‌dosage regimen‌

• ‌Basic PlanThree intraperitoneal injections (50 μ g/kg) per week for 6-7 weeks
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• ‌Strengthening planCombination of LPS (10-15 mg/kg) exacerbates fibrosis, and LPS is added after the last injection of huperzine
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• ‌Acute to chronicSimulate chronic disease progression through multiple acute inducements (such as weekly injections of huperzine)
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3、 Model evaluation indicators

1. ‌Pathological observation‌

• ‌H&E stainingReduction of acinar cells, infiltration of inflammatory cells, and interstitial fibrosis.

• ‌collagen depositionKu Wei Acid Sirius Red Staining Quantitative Fibrosis Degree
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• ‌immunohistochemistryDetecting α - SMA expression to reflect pancreatic stellate cell activation
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2. ‌biochemical indicators‌

• Serum levels of starch fixing enzyme and lipase
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• Detection of inflammatory factors (IL-6, IL-1 β, TNF - α)
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3. ‌Molecular mechanism analysis‌

• Western blot detection of NF - κ B/p65, IL-6 and other pathway proteins
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• QPCR analysis of fibrosis related genes (such as Smad3)

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