摘要
目的:探讨骨痹通消颗粒通过调控Wnt/PI3K⁃AKT信号通路对兔激素性股骨头坏死的治疗作用。方法:将30只清洁级新西兰兔随机分为对照组、模型组和实验组,除对照组外,其余均采用异体血清联合糖皮质激素建立模拟激素性股骨头坏死动物模型。造模成功后,模型组与对照组给予等量生理盐水,实验组按照人与兔的等效剂量给予骨痹通消颗粒浓缩液,1天1次,持续8周,经腹腔麻醉后体外心脏采血并采用免疫酶联吸附法检测血清中ALP、TGF⁃β1水平;处死实验动物,取出各组兔双侧股骨头进行苏木精⁃伊红染色,TUNEL染色法观察股骨头区细胞凋亡情况;RT⁃qPCR分析Bax、Bcl⁃2基因表达情况,Western blot检测通路蛋白Wnt、β⁃catenin、GSK⁃3β、AKT和p⁃AKT等表达水平。结果:与对照组相比,模型组兔骨组织中ALP和TGF⁃β1表达水平显著下降(P<0.05),骨小梁稀疏变细,空骨陷窝数和凋亡阳性细胞数明显增多(P<0.05),骨组织中促凋亡基因Bax表达升高,抗凋亡基因Bcl⁃2表达则受到抑制(P<0.05);经骨痹通消颗粒干预治疗后,实验组兔骨组织空骨陷窝情况和细胞凋亡情况均得到有效改善(P<0.05)。此外,与模型组对比,骨痹通消颗粒能够上调ALP和TGF⁃β1表达量(P<0.05),同时促进通路蛋白Wnt、β⁃catenin表达,提高AKT磷酸化水平,下调GSK⁃3β的表达(P<0.05)。结论:糖皮质激素环境下,成骨细胞发育功能障碍,细胞凋亡增多。骨痹通消颗粒能够调控Wnt/PI3K⁃AKT两条信号通路,促进成骨细胞发育并抑制细胞凋亡,有效维持骨代谢平衡,延长股骨头塌陷时间,进一步治疗股骨头坏死。
Objective:To investigate the therapeutic effect of osteopathic osteopathy on hormonal osteonecrosis in rabbits by regulating the Wnt/PI3K⁃AKT signaling pathway.Methods:Thirty clean⁃grade New Zealand rabbits were randomly divided in⁃to control group,model group and experimental group,and all except for the control group,allogeneic serum combined with glu⁃cocorticoids were used to establish animal models of simulated hormonal osteonecrosis of the femoral head.After successful model⁃ing,the experimental group was given the model group and the control group the same amount of normal saline,and the experi⁃mental group was given bone palsy granule concentrate according to the equivalent dose of humans and rabbits,once a day for 8 weeks,and after intraperitoneal anesthesia,blood was collected from the extracorporeal heart and the serum ALP and TGF⁃β1 levels were detected by immunoenzyme⁃linked adsorption.The experimental animals were sacrificed,and the bilateral femoral heads of rabbits were removed for hematoxylin⁃eosin staining,and the apoptosis of cells in the femoral head area was observed by TUNEL staining.RT⁃qPCR analyzed the expression of Bax and Bcl⁃2 genes,and Western blot detected the expression levels of pathway proteins Wnt,β⁃catenin,GSK⁃3β,AKT and p⁃AKT.Results:Compared with the control group,the expression levels of ALP and TGF⁃β1 in rabbit bone tissue in the model group decreased significantly(P<0.05),the bone trabeculae were sparse and thinned,the number of hollow bone fosses and the number of apoptosis⁃positive cells increased significantly(P<0.05),the expression of pro⁃apoptotic gene Bax was increased,and the expression of anti⁃apoptotic gene Bcl⁃2 was inhibited(P<0.05).Af⁃ter the intervention and treatment of bone palsy granules,the hollow bone depression and apoptosis of rabbit bone tissue in the ex⁃perimental group were effectively improved(P<0.05).In addition,compared with the model group,the osteopathic elimination particles could upregulate the expression of ALP and TGF⁃β1(P<0.05),promote the expression of pathway proteins Wnt andβ⁃catenin,increase the phosphorylation level of AKT,and downregulate the expression of GSK⁃3β(P<0.05).Conclusion:Un⁃der glucocorticoids,osteoblast development dysfunction and apoptosis increase.Bone palsy granules can regulate the two signaling pathways of Wnt/PI3K⁃AKT,promote osteoblast development and inhibit apoptosis,effectively maintain bone metabolic bal⁃ance,prolong the collapse time of the femoral head,and further treat osteonecrosis of the femoral head.
作者
李胜
朱彩玉
顾一帆
朱磊
李子鹏
陈少奇
周正新
LI Sheng;ZHU Cai-yu;GU Yi-fan;ZHU Lei;LI Zi-peng;CHEN Shao-qi;ZHOU Zheng-xin(First Clinical Medical College of Anhui University of Traditional Chinese Medicine,Hefei 230031,China;The First Affiliated Hospital of Anhui University of Traditional Chinese Medicine,Hefei 230031,China)
出处
《海南医学院学报》
CAS
2023年第7期523-529,共7页
Journal of Hainan Medical University
基金
安徽省高校研究生科研项目(YJS20210477)
安徽省自然科学基金面上项目(2008085MH281)
安徽省高校自然科学研究重点项目(KJ2020A0403)。
关键词
股骨头坏死
骨痹通消颗粒
糖皮质激素
成骨细胞
GSK⁃3β
Osteonecrosis of the femoral head
Bone palsy eliminates granules
Glucocorticoids
Osteoblasts
GSK⁃3β
作者简介
李胜,E⁃mail:1254045443@qq.com。;通讯作者:周正新,博士,主任医师,博士生导师,E⁃mail:zhouzhengxin1968@sina.com。
