The Expression Characteristics Of Nuclear Factor KappaB And Interleukin-6 In Mice Bone Tissue Of Experimental Osteoporosis Models

ObjectiveWith the population aging, the morbility of osteoporosis gradually step up and become an increasingly important public healthy problem. However due to the pathogenesy are not completely clear , its prevention and therapy are still difficult. The risk of fracture increased and the union of fracture prolonged. Because of the abnormal of molecular mechanism in bone microenvironment, we are paying more attention to the research of the role of cytokines in bony remodeling . In vitro research indicate that nuclear factor - kappaB can activate the transcription of IL - 6 and stimulate the differentiation of osteoclast. It will increase the number of osteoclast and the action of bone absorption. To examine the expression level of NF - kappaBp65 and Interleukin - 6, adult mice were ovariectomized as models of experimental osteoporosis. In this way , we can determin the protein level of NF - kappaBp65 and Interleukin - 6 , explore the molecular mechanism of osteoporosis and understand the relationship and significance between them during the morbility of osteoporosis. Deep reseach in the change of cytokines of bone microenvironment and explore to use molecular biology technique to improve it. That will possess important significance in the therapy of osteoporosis.Methods1. The grouping and duplication of osteoporosisFour - month female balb/c mice were randomly divided into ovariecto-mized group and sham - operated group.2. The establishment of osteoporosis model through Dual -X ray absorption method12 weeks after surgery ,the bone density of mice were measured by the e-quipment of Lunar bone density .3. The immunohistochemistry stain and in - situ hybridization detection The NF - kappaB and IL - 6 levels in bone tissue were determined by immunohistochemistry techniques. After sacrifice of the animal ,we get the inferior extremity of femur by sterile operation and set in the paraformaldehyde, perform immunohistochemistry stain and observation. Record probe by random priming, use Digoxin labeled NF - KBP65cDNA probe , perform according to the direction of the kit .4. Result assessment and image analysisThe bone density , NF - kappaB and IL - 6 levels of mice were demonstrate by average ± standard deviation and analyzed by t test and correlation a-nalysis.Results1. The bone density and histology observationCompared with the sham - operated group, we may find changes in the ovariectomized group: decrease in bone density , rarefaction of trabecula , the width of trabecula narrowing, interval among trabecula broadening.2. The immunohistochemistry stain and observationThe immunohistochemistry results showed that the positive signals of NF -kappaB and EL - 6 appeared similarly in both ovariectomized and sham - operated group. They mainly distribute in the cytoplasm of osteoblast, osteocytes, and marrow stroma cell.3. The result of immunohistochemistry stainThe content of protein in ovariectomized group are higher than in sham -operated group.4. In - situ hybridization observation and detection resultThe positive signal of NF - KBP65mRNA were found in both groups, they...