Application of a kind of amygdalin in the preparation of anti-inflammatory drugs for knee osteoarthritis
A technology for knee osteoarthritis and amygdalin is applied in the application field of amygdalin in the preparation of chondrocyte anti-inflammatory drugs to achieve the effects of enhancing anti-inflammatory ability, controlling side effects, and delaying degeneration
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Embodiment 1
[0034] Example 1 Expression characteristics of DNMT3B in human osteoarthritis cartilage
[0035] 1. Comparing the expression of DNMT3B in articular cartilage of non-diseased healthy people and OA patients
[0036] 1.1 Experimental samples: Normal articular cartilage tissue was collected from amputated patients' articular cartilage tissue (11 cases of talus or knee joint), and knee cartilage tissue of OA patients was collected from degenerated knee articular cartilage tissue under surgical replacement after total knee arthroplasty ( 57 cases).
[0037] 1.2 Experimental grouping
[0038] (A) Non-disease (ND) group; (B) OA group.
[0039] 1.3 Acquisition of articular cartilage tissue and preparation of slices
[0040] After the samples of each group were washed several times with HBSS (Hank's Balanced Salt Solution), they were soaked in 10% NBF (prepared with ultrapure water) for 3 days to fully fix them. After fixation, rinse with deionized water 6 times (15 minutes / time), t...
Embodiment 2
[0116] Example 2 In vitro study of amygdalin delaying chondrocyte degeneration by regulating Dnmt3b
[0117] 1. Method
[0118] 1. Screening of optimal drug concentration for AMD in chondrocytes
[0119] 1.1 Experimental grouping
[0120] (A) blank group; (B) control group; (C) drug group.
[0121] 1.2 Isolation and culture of mouse articular chondrocytes
[0122] 1) Take 2-week-old C57BL / 6J mice and use CO 2 After sacrifice, soak them in 70% alcohol for 5-10 minutes.
[0123] 2) Take out the mouse, peel off the skin, grab the cartilage layer of the femoral head with tweezers, cut off the attached ligament and soft tissue with scissors, and place the cartilage tissue in a 1.5 mL centrifuge tube containing DPBS.
[0124] 3) Remove the absorbent cloth, clean the ultra-clean workbench, change gloves, and preheat the digestion medium (DMEM medium + 10% FBS + 1% P / S + collagenase P 0.5 mg / mL) at 37°C.
[0125] 4) Wash the cartilage cap in the 1.5mL centrifuge tube with DPBS 4...
Embodiment 3
[0169] Example 3 In vitro study of amygdalin preventing chondrocyte degeneration by regulating p38 / MAPK / Dnmt3b signaling pathway
[0170] Objective To clarify the changes of inflammation-related pathways in chondrocytes regulated by AMD in the inflammatory state, and to clarify the mechanism of AMD preventing chondrocyte degeneration.
[0171] 1. Method
[0172] 1. To clarify the regulatory effect of AMD on inflammation-related pathways in chondrocytes in the inflammatory state
[0173] 1.1 Experimental grouping
[0174] (A) Veh group; (B) IL-1β group; (C) AMD group; (D) IL-1β+AMD group.
[0175] 1.2 Isolation and culture of mouse articular chondrocytes
[0176] Articular chondrocytes were isolated from knee joints of 2-week-old C57BL / 6J mice. Separation method method is the same as embodiment 2 experimental method 2.2. Take 2.5×10 5 / mL density was inoculated into tissue culture plates containing basal medium (DMEM medium+10%FBS+1%P / S).
[0177] 1.3 Inflammatory factor...
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