葛根素调控miR-30b-5p表达抑制类固醇诱导的兔股骨头坏死

Puerarin regulates miR-30b-5p expression and inhibits steroid induced necrosis of the femoral head in rabbits

目的探讨葛根素调控miR-30b-5p表达对类固醇诱导的兔股骨头坏死(osteonecrosis of the femoral head,ONFH)的作用及机制。方法将84只新西兰大白兔分为对照组、ONFH组、葛根素低剂量组、葛根素高剂量组、阳性对照组、葛根素高剂量+antagomir NC组、葛根素高剂量+miR-30b-5p antagomir组,每组12只。通过Micro-CT检测大白兔股骨头骨小梁数量、骨小梁厚度及骨小梁分离度;HE染色检测股骨头组织病理变化;ELISA法检测股骨头组织中超氧化物歧化酶(SOD)活性及丙二醛(MDA)含量;TUNEL染色检测股骨头组织中骨细胞凋亡;qRT-PCR检测股骨头组织中miR-30b-5p表达;Western Blot检测股骨头组织中Dickkopf-1(DKK-1)蛋白表达。结果与对照组比较,ONFH组骨小梁数量、骨小梁厚度、SOD活性、miR-30b-5p表达降低,骨小梁分离度、空骨陷窝率、MDA含量、骨细胞凋亡率及DKK-1蛋白表达升高(P<0.05);与ONFH组比较,葛根素低、高剂量组、阳性对照组骨小梁数量、骨小梁厚度、SOD活性、miR-30b-5p表达升高,骨小梁分离度、空骨陷窝率、MDA含量、骨细胞凋亡率及DKK-1蛋白表达降低(P<0.05);miR-30b-5p antagomir减弱了高剂量葛根素对ONFH兔氧化应激及细胞凋亡的抑制作用。结论葛根素可抑制氧化应激及细胞凋亡进而发挥对ONFH兔的保护作用,其机制可能与上调miR-30b-5p表达有关。

Objective To investigate the effect and mechanism of puerarin on steroid-induced osteonecrosis of the femoral head(ONFH)in rabbits by regulating the expression of miR-30b-5p.Methods A total of 84 New Zealand white rabbits were grouped into a control group,ONFH group,low-dose puerarin group,high-dose puerarin group,positive control group,high-dose puerarin+antagonist NC group,and high-dose puerarin+miR-30b-5p antagonist group,with 12 rabbits in each group.Micro CT was applied to detect the number,thickness,and separation of trabeculae in the femoral head of rabbits;HE staining was applied to detect pathological changes in the femoral head tissue;ELISA was applied to detect the activity of superoxide dismutase(SOD)and the content of malondialdehyde(MDA)in the femoral head tissue;TUNEL staining was applied to detect osteocyte apoptosis in femoral head tissue;qRT-PCR was applied to detect the expression of miR-30b-5p in femoral head tissue;Western blot was applied to detect the expression of Dickkopf-1(DKK-1)protein in femoral head tissue.Results Compared with the control group,the number of trabeculae,thickness of trabeculae,SOD activity,and the expression of miR-30b-5p in the ONFH group decreased,the bone trabecular separation,empty bone lacuna rate,MDA content,bone cell apoptosis rate,and the expression of DKK-1 protein increased(P<0.05);compared with the ONFH group,the number of trabeculae,thickness of trabeculae,SOD activity,and the expression of miR-30b-5p in the low-dose,the high-dose puerarin group,and the positive control group increased,the bone trabecular separation,empty bone lacuna rate,MDA content,bone cell apoptosis rate,and the expression of DKK-1 protein decreased(P<0.05);miR-30b-5p antagomir attenuates the inhibitory effect of high dose puerarin on oxidative stress and apoptosis of ONFH rabbits.Conclusion Puerarin can inhibit oxidative stress and apoptosis of ONFH rabbits,and its mechanism may be related to the up-regulation of miR-30b-5p expression.