Snow Glycosides Model Mouse Fibroblasts To Inhibition By Light Microscopy Electron Microscopy Experiments

1.Objective:The experiment is to discuss the mechanism of asiaticoside treat fibroblast after inflammation from the animal model. To understanding the change of ultramicrostructure of fibroblast in the inflammation developing process through observating the light mirror and the electricity mirror ,providing the theoretical basis for the asiaticoside in the clinical application. 2.Experimental material:2.1 Medicines and reagent: The asiaticoside, the Shanghai modern Pudong pharmaceutical factory limited company produces, batch number 050401, specification 6mg/Piece; Tetracyclin, Chengdu Jinghua pharmaceutical manufacturing plant production, batch number 050302, specification 0.25g/ Piece; Propionibacterium acne , provides by the Australian bacterium laboratory; 10% formaldehyde solution, provide by the Wuhan University medical school immunity room; 2.5% glutaric dialdehydes provide by the Wuhan University medical school electricity mirror room.2.2 Experimental animals: Kunming pure line mouse 40, female and male are half and half, body weight 28±2g,The mouse age 2 months, provide by the Huazhong science and technology University Tongji medical school experimental animal department2.3 Instruments: Photoelectricity binocular microscope 1, provides by the Wuhan University medical school immunity pathology laboratory; Cell counter 1, provides by the Wuhan University medical school immunity pathology laboratory; LKB2V ultra thin slicer slice,Hitachi H2600 passes the radio mirror to provide by the Wuhan University medical school electricity mirror laboratory.3.Methods:3.1 Propionibacterium acnes culture: The freezing and drying Propionibacterium acnes dissolutes in the brain heart infusion,then layes aside several minutes under the room temperature , takes the little solution culture in 5% blood agar, tires of the oxygen environment in the middle of 35-37℃ to culture for 5-7 day, washes 3 times with the physiological saline,adjusts Propionibacterium acnes concertration for 2 × 10¹⁰/ml spare, and through light mirror confirms that it's blue rod-shaped Propionibacterium acnes.3.2 Animal model: shearing fur of the mouse back, approximately 1×1 centimeter size, each mouse three sides;then injecting cultured Propionibacterium acnes into the mouse subsurface,each side injects 20 million units Propionibacterium acnes,and observe the change of mouse partial skin after injecting. <> Chinese medicine publishing house 256)3.3 Batching:Dividing the experiment mouse into four groups:the asiaticoside and tetracyclin group,tetracyclin group, asiaticoside group,Tetracyclin group; blank Contrast group.3.4 Intragastral:The asiaticoside+tetracyclin group gives asiaticoside 0.25ml (0.144mg/ml) and the tetracyclin 0.25ml (1.5mg/ml),The asiaticoside group givesasiaticoside0.25ml (0.144mg/ml),The tetracyclin group gives tetracyclin 0.25ml (1.5mg/ml) , blank Contrast group gives physiological saline 0.25ml. According to stipulation dosage gives the medicine.in the morning 8-10 of every day. <> Hubei science and technology publishing house 277 commonly used experimental animals and human's body surface area proportion)3.5 Light mirrors specimen manufacture and observation:excising the mice skin in 3, 7, 10, 14, 21 days.Fixed with 20% formaldehyde.According to paraffin section...