摘要
背景:前期研究发现,对于绝经后骨质疏松症患者,补肾健脾方有助于调节血清破骨、成骨相关因子及肠道菌群水平,改善患者临床症状并进一步提高疗效。目的:观察补肾健脾方对去势大鼠骨代谢及骨微结构的影响。方法:采用随机数字表法将24只雌性SD大鼠随机分为4组:假手术组(n=6)切除卵巢周围脂肪,模型组(n=6)、阿仑膦酸钠组(n=6)、补肾健脾方组(n=6)切除双侧卵巢建立去势模型;造模5 d后,补肾健脾方组予以补肾健脾方灌胃(1次/d),阿仑膦酸钠组予以阿仑膦酸钠灌胃(每周1次),假手术组及模型组给予等体积生理盐水灌胃(1次/d),连续灌胃12周。灌胃结束后,检测血清Ⅰ型胶原氨基端延长肽、Ⅰ型胶原羟基端肽β降解产物及肿瘤坏死因子α水平,Micro-CT检测股骨远端微结构,苏木精-伊红染色观察股骨组织形态,免疫组化染色检测股骨内Ⅰ型胶原蛋白、肿瘤坏死因子α表达。结果与结论:①血清ELISA检测显示,与假手术组比较,模型组Ⅰ型胶原氨基端延长肽水平降低(P<0.05),Ⅰ型胶原羟基端肽β降解产物、肿瘤坏死因子α水平升高(P<0.05);与模型组比较,补肾健脾方组与阿仑膦酸钠组Ⅰ型胶原氨基端延长肽水平升高(P<0.05),Ⅰ型胶原羟基端肽β降解产物、肿瘤坏死因子α水平降低(P<0.05);②Micro-CT三维重建显示,与假手术组对比,模型组骨小梁数量显著减少、连续性变差、间隙增大,阿仑膦酸钠与补肾健脾方均对此种现象有积极改善作用;③苏木精-伊红染色显示,假手术组比较,模型组骨小梁数量减少且形态变细,网状结构遭到破坏;与模型组比较,补肾健脾方组与阿仑膦酸钠组骨小梁数量增多且形态变粗,网状结构有所恢复;④免疫组化染色显示,与假手术组比较,模型组Ⅰ型胶原蛋白表达减少,肿瘤坏死因子α表达增加;与模型组相比,补肾健脾方组与阿仑膦酸钠组Ⅰ型胶原蛋白表达增加,肿瘤坏死因子α表达减少。结果表明,补肾健脾方可有效改善去势大鼠的骨代谢及骨微结构。
BACKGROUND:Previous studies have found that for postmenopausal osteoporosis patients,the Bushen Jianpi Formula can help regulate serum osteoclast-and osteoblast-related factors and intestinal flora levels,improve clinical symptoms of patients and further improve efficacy.OBJECTIVE:To investigate the effect of Bushen Jianpi Formula on bone metabolism and bone microarchitecture in ovariectomized rats.METHODS:Twenty-four female SD rats were randomly divided into four groups by random number table method:sham operation group(n=6)was subjected to periovarian fat removal,and bilateral ovariectomy was performed in model group(n=6),alendronate group(n=6),and Bushen Jianpi Formula group(n=6).Five days after modeling,the Bushen Jianpi Formula group was given Bushen Jianpi Formula by intragastric administration(once a day).The alendronate sodium group was given alendronate by intragastric administration(once a week).The sham operation group and model group were given an equal volume of normal saline by intragastric administration(once a day)for 12 consecutive weeks.After intragastric administration,serum levels of procollagen I N-terminal peptide,β-isomerized C-terminal telopeptide of type I collagen degradation product,and tumor necrosis factorαwere detected.The distal femur microstructure was detected by Micro-CT.The femoral tissue morphology was observed by hematoxylin-eosin staining.The expression of type I collagen and tumor necrosis factorαin the femur was detected by immunohistochemical staining.RESULTS AND CONCLUSION:(1)The serum ELISA test showed that compared with the sham operation group,the level of procollagen I N-terminal peptide in the model group decreased(P<0.05),and the levels of degradation products ofβ-isomerized C-terminal telopeptide of type I collagen and tumor necrosis factorαincreased(P<0.05).Compared with the model group,the levels of the procollagen I N-terminal peptide in the Bushen Jianpi Formula group and the alendronate sodium group increased(P<0.05),and the levels ofβ-isomerized C-terminal telopeptide of type I collagen degradation products and tumor necrosis factorαdecreased(P<0.05).(2)Micro-CT three-dimensional reconstruction showed that compared with the sham operation group,the number of bone trabeculae in the model group was significantly reduced,the continuity was worsened,and the gaps were increased.Both alendronate sodium and Bushen Jianpi Formula had a positive improvement effect on this phenomenon.(3)Hematoxylin-eosin staining showed that compared with the sham operation group,the number of bone trabeculae in the model group was reduced and the shape became thinner,and the network structure was destroyed.Compared with the model group,the number of bone trabeculae in the Bushen Jianpi Formula group and the alendronate sodium group increased,the shape became thicker,and the reticular structure was restored.(4)Immunohistochemical staining showed that compared with the sham operation group,the expression of type I collagen was reduced and tumor necrosis factorαexpression was increased in the model group.Compared with the model group,the expression of type I collagen increased and the expression of tumor necrosis factorαdecreased in the Bushen Jianpi Formula group and the alendronate sodium group.The results show that the Bushen Jianpi Formula can effectively improve bone metabolism and bone microstructure in ovariectomized rats.
作者
郭荪林
洪恩达
戴新华
林禧
彭志毅
程英雄
范琳燕
Guo Sunlin;Hong Enda;Dai Xinhua;Lin Xi;Peng Zhiyi;Cheng Yingxiong;Fan Linyan(Third Clinical Medicine College of Guangzhou University of Chinese Medicine,Guangzhou 510405,Guangdong Province,China;Third Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510375,Guangdong Province,China;Guangzhou University of Chinese Medicine,Guangzhou 510006,Guangdong Province,China)
出处
《中国组织工程研究》
北大核心
2025年第26期5588-5594,共7页
Chinese Journal of Tissue Engineering Research
基金
广东省自然科学基金面上项目(2021A1515011463),项目负责人:范琳燕。
关键词
补肾健脾方
去势大鼠
骨质疏松
骨代谢
骨微结构
MICRO-CT
工程化组织构建
Bushen Jianpi Formula
ovariectomized rat
osteoporosis
bone metabolism
bone microarchitecture
Micro-CT
engineered tissue construction
