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The Intervention Role of Ground Beetle on Steroid-induced BMSCs Adipogenic Differentiation

Author: LiShuQiang
Tutor: QiZhenXi
School: Fujian College of Traditional Chinese Medicine
Course: TCM orthopedics and traumatology
Keywords: Mesenchymal Stem Cells / drug effects Spinal cord Necrosis of the femoral head / TCD therapy Eupolyphaga Cell differentiation / drug effects Cell culture technology
CLC: R285.5
Type: Master's thesis
Year: 2009
Downloads: 87
Quote: 1
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Abstract


Objective: steroid-induced avascular necrosis may be induced with hormone adipogenic differentiation of mesenchymal stem cells (BMSCs) in bone marrow, Eupolyphaga significant effect on the prevention and treatment of the disease early. This research project by observing the hormone-induced rat bone marrow mesenchymal stem cells the adipogenic differentiation Eupolyphaga containing serum explore the Eupolyphaga hormone induced adipogenic differentiation of mesenchymal stem cells in the bone marrow of the impact of its intervention role for clinical prevention hormone The avascular necrosis laid a theoretical foundation to provide a theoretical basis for the treatment of this disease and is Chinese medicine Eupolyphaga on. Methods: (1) selection quality Chinese medicine Eupolyphaga using the the ten days Preparation Eupolyphaga containing serum. (2) to take the 1-month-old SD rats tibia and femur bone marrow BMSCs adherent culture and subculture. The the detected the biography three generations BMSCs surface antigens CD45 and CD90 expression positive rate to identify. (3) pass three generations of BMSCs were randomly divided into blank group, the control group and Chinese medicine is low, medium and high dose groups. Control group, a high dose of hormones induced it to fat cells, traditional Chinese medicine each group to give Eupolyphaga hormone induced serum containing intervention. (4) intervention in each group after 6 days of osteogenic and adipogenic marker testing, including an inverted microscope, alkaline phosphatase (ALP), triglycerides (TG), osteocalcin (BGP) detection, RT- PCR of type I collagen (Col I), the fat cells of peroxisome proliferator-activated receptor (PPARγ) and fatty acid binding protein (aP2) mRNA expression of determination. Results: 1. According BMSCs adherent growth characteristics of BMSCs and hematopoietic cells in the bone marrow can be separated. Isolation and culture through such methods BMSCs surface antigen CD45, the CD90 expression positive rate of 1.9% and 99.1%, respectively. Cell intervention after 6 days of culture triglyceride content in the cells of the control group than the control group was significantly higher, and the difference was highly significant (P <0.01). Chinese medicine groups than in the control group were decreased to varying degrees, and the control group showed a highly significant (P <0.01). With the the Chinese medicine concentration increased intracellular triglyceride content showed a decreasing trend. Cell intervention after 6 days of culture, control group culture fluid in the osteocalcin content lower than the control group, the difference was highly significant (P <0.01). Chinese medicine group than in the control group had varying degrees of rise, and Chinese medicine concentration increased osteocalcin levels in the culture fluid showed an increasing trend. Chinese herbal medicine, the high-dose group and control group difference was highly significant (P <0.01). Cell intervention after 6 days of culture, alkaline phosphatase (ALP) levels in the cells of the control group were significantly lower than the blank, and the difference was highly significant (P <0.01). Chinese medicine group than in the control group has increased to varying degrees, and the control group showed a highly significant (P <0.01). With the the Chinese medicine concentration increased intracellular ALP content showed an increasing trend. 5 gene expression: cell intervention after 6 days of culture, semi-quantitative analysis of PCR products gel imaging scan system, collagen type I, fat cells peroxisome proliferator-activated receptor (PPARγ) and fatty acid binding protein (aP2) mRNA expression in the control group, blank group and Chinese medicine group showed a significant difference. Conclusion: 1. Experiments in vitro were isolated and cultured cells by morphology, flow cytometry surface markers identified belong BMSCs, and has a higher purity. . Induced by high-dose hormone may promote BMSCs differentiation to adipogenic and osteogenic differentiation to reduce. 3 Eupolyphaga serum containing intervention intracellular triglyceride the content, PPARγmRNA and aP2 mRNA expression compared with the control group were significantly lower adipogenic differentiation of BMSCs hormone induced the prompt Eupolyphaga reversible. 4. The serum containing Eupolyphaga intervention BMSCs, the expression of cell ALP, BGP content and Col ImRNA, compared with the control group increased in the prompt Eupolyphaga promote BMSCs osteogenic differentiation. 5. Eupolyphaga prevention mechanism of steroid-induced avascular necrosis is not only to improve the microcirculation of the femoral head, also inhibit BMSCs adipogenic differentiation, promote osteogenic differentiation.

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