Experimental method of effects of DNA oxidative damage and bone cell apoptosis on avascular necrosis

Abstract

The invention discloses an experimental method of effects of DNA oxidative damage and bone cell apoptosis on avascular necrosis, the method comprises the following steps: experimental grouping and establishment of an animal model, preparation of specimens, histomorphology observation under a light microscope, observation by a transmission electron microscope, bone cell apoptosis situation detection by TUNEL (terminal dexynucleotidyl transferase(TdT)-mediated dUTP nick end labeling) method, bone marrow hematopoietic cell DNA oxidative damage detection by monoclonal antibody N45.1 immunohistochemistry method, and statistical analysis. According to the method, experiment animals are respectively killed in the second week and the fourth week after the last time of administration, and five animals are killed for each time in each group; bilateral femoral heads are taken for conventional fixed decalcification for stand-by testing, the number of vacant bone lacunas is counted by HE (hematoxylin-eosin) staining, the bone cell morphological change is observed by the electron microscope, the bone cell apoptosis situation is detected by the TUNEL method, the bone marrow hematopoietic cell DNA oxidative damage is detected by the immunohistochemistry method, and the relationship between SANFH ((steroid-induced avascular necrosis of femoral head)) and the bone marrow hematopoietic cell DNA oxidative damage and the bone cell apoptosis is explored, and provides new ideas for the further study of pathogenesis of the steroid induced avascular necrosis of femoral head.

Description

technical field [0001] The invention belongs to the research field of femoral head necrosis, and particularly relates to an experimental method for the effect of hematopoietic cell DNA oxidative damage and osteocyte apoptosis in ischemic necrosis. Background technique [0002] Steroid-induced Avascular Necrosis of Femoral Head (SANFH) refers to partial or complete ischemia of the femoral head caused by long-term use or short-term massive use of adrenal cortex hormones (hereinafter referred to as hormones), which in turn leads to bone The pathological process of necrosis of cells, bone marrow hematopoietic cells and fat cells has been recognized by the medical community. Since it was first reported in 1957, there have been many clinical cases. [0003] SANFH mainly affects young and middle-aged people, and is mostly bilateral, with a wide range of necrosis. After the "SARS" in 2003, a large number of cases emerged again. Hormone-induced SANFH is a complex biological proces...

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Problems solved by technology

[0010] The purpose of the present invention is to provide an experimental method for the role of hematopoietic DNA oxidative damage and osteocyte apoptosis in ischemic necrosis, aiming to solve what kind of pathological changes occur in the early stage of SANFH, whether there is DNA oxidative damage, the pathological changes and What is the temporal relationship between the occurrence of osteocyte apoptosis

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