To investigate the effect of calcitonin gene related peptide (CGRP) on bone resorption mediated by interleukin 1β(IL 1β) in vitro , the osteoclasts isolated from the long bones of newborn SD rats were co cul...To investigate the effect of calcitonin gene related peptide (CGRP) on bone resorption mediated by interleukin 1β(IL 1β) in vitro , the osteoclasts isolated from the long bones of newborn SD rats were co cultured with osteoblasts on ivory slices placed in 24 well plates . 24 h later, conditioned media containing CGRP and/or IL 1β were added to the wells respectively, and continued culturing for 48 h. After the cells were stripped off by ultrasonication, the ivory slices were stained in toludine blue. The number and the total area of resorption lacunae on each slice were measured by computer imaging analysis system. Our results showed that IL 1β significantly stimulated bone resorption, but CGRP inhibited the effect mediated by IL 1β in a dose dependent manner. It is suggested that CGRP may inhibit osteoclastic bone resorption through two ways: One is that CGRP functions directly on osteoclasts to block their activation; the other is that CGRP regulates the release of cytokines by osteoblasts and indirectly affects the function of osteoclasts.展开更多
目的探究柚皮苷对破骨细胞分化和极化功能的影响。方法体外培养RAW264.7细胞,设置对照组(培养基不含柚皮苷)和实验组(培养基中柚皮苷剂量分别为2、20、200μg/L)连续培养7 d,通过抗酒石酸酸性磷酸酶(TRAP)染色对比组间诱导分化结果,通...目的探究柚皮苷对破骨细胞分化和极化功能的影响。方法体外培养RAW264.7细胞,设置对照组(培养基不含柚皮苷)和实验组(培养基中柚皮苷剂量分别为2、20、200μg/L)连续培养7 d,通过抗酒石酸酸性磷酸酶(TRAP)染色对比组间诱导分化结果,通过实时荧光定量逆转录聚合酶链反应(qPCR)对比组间组织蛋白酶K(CK)、基质金属蛋白酶-9(MMP-9)、整合素β3(integrinβ3)和非受体酪氨酸激酶(c-src)mRNA表达水平,通过蛋白免疫印迹法对比组间CK、MMP-9、integrinβ3、磷酸化产物p-src蛋白表达水平。结果与对照组相比,各实验组TRAP染色阳性细胞数显著降低(P<0.05);与对照组相比,20μg/L与200μg/L组CK、MMP-9、integrinβ3及c-src m RNA表达水平显著降低(P<0.05);与对照组相比,20μg/L与200μg/L组CK、MMP-9、integrinβ3及p-src蛋白表达水平显著降低(P<0.05)。结论柚皮苷对破骨细胞的分化和极化有一定的抑制作用,其作用机制可能与下调integrinβ3、c-src及p-src表达有关。展开更多
文摘To investigate the effect of calcitonin gene related peptide (CGRP) on bone resorption mediated by interleukin 1β(IL 1β) in vitro , the osteoclasts isolated from the long bones of newborn SD rats were co cultured with osteoblasts on ivory slices placed in 24 well plates . 24 h later, conditioned media containing CGRP and/or IL 1β were added to the wells respectively, and continued culturing for 48 h. After the cells were stripped off by ultrasonication, the ivory slices were stained in toludine blue. The number and the total area of resorption lacunae on each slice were measured by computer imaging analysis system. Our results showed that IL 1β significantly stimulated bone resorption, but CGRP inhibited the effect mediated by IL 1β in a dose dependent manner. It is suggested that CGRP may inhibit osteoclastic bone resorption through two ways: One is that CGRP functions directly on osteoclasts to block their activation; the other is that CGRP regulates the release of cytokines by osteoblasts and indirectly affects the function of osteoclasts.
文摘目的探究柚皮苷对破骨细胞分化和极化功能的影响。方法体外培养RAW264.7细胞,设置对照组(培养基不含柚皮苷)和实验组(培养基中柚皮苷剂量分别为2、20、200μg/L)连续培养7 d,通过抗酒石酸酸性磷酸酶(TRAP)染色对比组间诱导分化结果,通过实时荧光定量逆转录聚合酶链反应(qPCR)对比组间组织蛋白酶K(CK)、基质金属蛋白酶-9(MMP-9)、整合素β3(integrinβ3)和非受体酪氨酸激酶(c-src)mRNA表达水平,通过蛋白免疫印迹法对比组间CK、MMP-9、integrinβ3、磷酸化产物p-src蛋白表达水平。结果与对照组相比,各实验组TRAP染色阳性细胞数显著降低(P<0.05);与对照组相比,20μg/L与200μg/L组CK、MMP-9、integrinβ3及c-src m RNA表达水平显著降低(P<0.05);与对照组相比,20μg/L与200μg/L组CK、MMP-9、integrinβ3及p-src蛋白表达水平显著降低(P<0.05)。结论柚皮苷对破骨细胞的分化和极化有一定的抑制作用,其作用机制可能与下调integrinβ3、c-src及p-src表达有关。