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Influence of Rosiglitazone on Both Bone Mineral Bensity and the Expression of BMP-2 and Bcl-xL in Osteoblast in OLETF Rats

Author: WuHaiQing
Tutor: LiangYu
School: Tianjin Medical University
Course: Human Anatomy and Embryology
Keywords: Rosiglitazone Type 2 diabetes mellitus Bone density OLETF rats Osteoblasts Bone morphogenetic protein-2 Collagen
CLC: R587.1
Type: Master's thesis
Year: 2011
Downloads: 40
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Abstract

The purpose of diabetes mellitus (DM), following heart disease, cancer, diseases of serious harm to people's health, with the advent of an aging society, the incidence of diabetes has increased year by year. Diabetic Osteoporosis (DOP) is a secondary bone osteoporosis (OP) is the most common complication of DM, accompanied by changes in nutritional, metabolic, endocrine factors and occurred a series of pathological changes. OLETF rats, a spontaneous type 2 diabetic rats, the experimental type 2 diabetes model OLETF rats as LETO rats and photo Ruluo Ge rosiglitazone OLETF rats as control, by observing the rat femur pathological changes measured The bone mineral density (BMD) and osteoblast bone morphogenetic protein -2 (BMP-2), Bcl-xL expression levels to explore type 2 diabetes on bone and rosiglitazone on diabetic rat bone and characteristics. Single cages under specific pathogen-free (SPF) conditions of class methods rats fed the standard diet, 12/12h light-dark cycle, free access to food and drinking water. An oral glucose tolerance test (OGTT) regularly monitor blood glucose the blood glucose peak gt; 16.7mmol / L and 120 min after load blood glucose gt; 11.1mmol / L, a diagnosis of diabetes, have only the one for impaired glucose tolerance. Into a mold OLETF rats 12 to 30 weeks, a total, were randomly divided into diabetic control (DM) group, rosiglitazone (RGZ) group (n = 6), 8 LETO rats as normal control (NC) group . RGZ group: Rosiglitazone was diluted with distilled water and was dissolved at a dose of 3mg/kg · d. DM group and NC group distilled water to the same amount in each group were administered orally for 12 weeks, 1 day. Rats were sacrificed, the rat was removed from the body of the left and right hind limbs femur carefully peeling the soft tissue adhered to the surface, to avoid damage bone surface. Left hind femur wrapped in gauze soaked with phosphate buffered saline and stored at -20 ℃ prepared to do detection of the BMD, collagen content and the ashes dry weight ratio. Right hind limb femur placed and fixed in 10% neutral formalin solution with 10? The TA decalcification prepared with HE and osteoblasts BMP-2, Bcl-xL immunohistochemical staining. Histomorphometric analysis application of multimedia analysis software SPSS13.0 software for statistical analysis. Results 1. Rats: Comparison of of OLETF rats compared with LETO and obese, sluggish activity, lazy move, mental Weidun, lack of exercise with eyes closed, arched back and curled up, hi get together, chills, warm, dry dull coat. The femur BMD compare: DM group [(0.198 ± 0.011) g/cm2] compared, NC group BMD [(0.220 ± 0.012) g/cm2] increased (P lt; 0.01), RGZ group BMD [(0.172 ± 0.012) g/cm2] lower (P lt; 0.01). Ashes dry weight ratio: compared with the DM group (0.521 ± 0.012) and NC group shares the ashes of dry weight ratio (0.538 ± 0.018) increased (P lt; 0.05), dry weight RGZ group (0.492 ± 0.011) shares ashes decreased (P lt; 0.01). 4. The decalcification situation femur comparison: compared with DM group, NC group femur decalcification time (bone - demineralized bone weight) / weight ratio increased bone compared to statistical significance (P lt; 0.01 ). RGZ group relative to the DM group decalcification time (bone - demineralized bone weight) / weight ratio decreased bone compared statistically significant (P lt; 0.05). 5 the empty osteocyte lacuna rate, counting change in osteoblast (OB) and osteoclasts (OC): Compared with DM group, NC group empty bone cells subsidence the fossa rate and OC counts lower (P lt; 0.01 or P lt; 0.05), OB count increased (P lt; 0.01). RGZ group relative to the DM group empty bone cells subsidence the nest rate and OC count increased (P lt; 0.01), OB count decreased (P lt; 0.05). 6. Femur collagen content comparison: compared with DM group [(185.60 ± 19.38) mg / g] NC group femur collagen content [(243.27 ± 22.19) mg / g] increased (P lt; 0.01), RGZ group [ (158.81 ± 17.65) mg / g lower (P lt; 0.05). 7 osteoblast cells Bcl-xL expression changes: compared with the DM group (84.83 ± 10.82), NC of osteoblasts positive Bcl-xL expression an average gray value (69.13 ± 11.56) reduce (P lt; 0.05), RGZ composed bone cells positive expression of Bcl-xL elevated average gray value (110.17 ± 15.33) (P lt; 0.01). Osteoblasts BMP-2 expression changes: RGZ composition the bone cells BMP-2 positive expression absorbance value (0.4247 ± 0.0321) was significantly lower than the DM group (0.4886 ± 0.0398), and the absorbance value of the positive expression of the DM group was significantly lower than NC group (0.5571 ± 0.0402), the results were statistically significant (P lt; 0.01). Conclusions The rat model significantly obese, can be used as an ideal animal model of type 2 diabetes, bone metabolism disorder. 2 diabetic state of rats showed reduced bone density and bone mass, rats may appear abnormal the apparent bone tissue morphology, osteoblast dysfunction may be the main mechanism of diabetic bone metabolic imbalance, but osteoclasts may also be involved them. The diabetic rat model abnormal collagen metabolism and structural disorder, the former showed a decrease in collagen synthesis and content, the latter showed poor continuity of bone collagen and disarranged. The collagen metabolic abnormalities may be related to the decline in the number of osteoblasts or dysfunction. 4. Rosiglitazone on spontaneous obese type 2 diabetic rat femur with aggravated damage prompted our clinical medication to pay attention to its harmful effects on bone.

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