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接骨膏促进新西兰兔骨折愈合的活性部位筛选
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郑国尧 ,彭 磊,徐元翠,吴 飞,吴 波(恩施土家族苗族自治州中心医院康复医学中心,湖北 恩施
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445099)
中图分类号 R965.1 文献标志码 A 文章编号 1001-0408(2023)11-1349-05
DOI 10.6039/j.issn.1001-0408.2023.11.12
摘 要 目的 筛选接骨膏促进新西兰兔骨折愈合的活性部位。方法 制备接骨膏醇提物以及乙酸乙酯部位、正丁醇部位,并与炼
蜜调制成黏稠度适宜的膏药。以新西兰兔建立左前肢桡骨骨折模型,然后分为模型对照组、醇提物组、乙酸乙酯部位组、正丁醇部
位组,每组6只。除模型对照组外,其余各组兔外敷接骨膏相应极性部位膏药,干预4周。通过X线检测兔骨折愈合情况,采用酶
联免疫吸附测定法检测兔血清中白细胞介素6(IL-6)、肿瘤坏死因子α(TNF-α)、骨钙素(OC)、血管内皮生长因子A(VEGFA)、碱
性成纤维细胞生长因子2(bFGF2)、碱性磷酸酶(ALP)水平,采用苏木素-伊红染色观察兔骨折部位病理学形态变化,采用免疫组
化法检测兔骨折部位bFGF2蛋白的表达水平。结果 正丁醇部位组兔骨折部位的愈合速度最快,醇提物组次之,乙酸乙酯部位组
最慢;正丁醇部位组兔血清中 TNF-α、IL-6 水平下降最快,ALP、bFGF2、OC、VEGFA 水平上升最快[且较醇提物组显著升高(P<
0.01)],骨折部位的软骨细胞完全消失,形成大量骨髓腔,且骨髓腔内骨小梁正式形成,bFGF2表达水平也较醇提物组升高。结论
接骨膏正丁醇部位促进兔骨折部位愈合的作用强于醇提物以及乙酸乙酯部位,是接骨膏促进骨折愈合的活性部位。
关键词 接骨膏;骨折;愈合;醇提物;正丁醇部位;活性部位
Screening of active fractions of Jiegu ointment for promoting fracture healing in New Zealand rabbits
ZHENG Guoyao,PENG Lei,XU Yuancui,WU Fei,WU Bo(Rehabilitation Medical Center, Central Hospital of
Enshi Tujia and Miao Autonomous Prefecture, Hubei Enshi 445099, China)
ABSTRACT OBJECTIVE To screen the active site of Jiegu ointment in promoting fracture healing in New Zealand rabbits.
METHODS The ethanol extract of Jiegu ointment, as well as the ethyl acetate and n-butanol parts, were prepared and mixed with
honey to form a plaster with appropriate viscosity. The radial fracture model of left forelimb in New Zealand rabbit was established
and divided into model control group, ethanol extract group, ethyl acetate fraction group and n-butanol fraction group, with 6
rabbits in each group. Except for model control group, rabbits of all other groups were treated with corresponding polar part of
Jiegu ointment for external application, for 4 weeks. The radial fracture healing of rabbits was studied by X-ray examination.
Enzyme-linked immunosorbent assay was used to detect the serum levels of interleukin-6 (IL-6), tumor necrosis factor α (TNF-α),
osteocalcin (OC), vascular endothelial growth factor A (VEGFA), basic fibroblast growth factor 2 (bFGF2) and alkaline
phosphatase (ALP). HE staining was adopted to observe the changes of pathological morphology of rabbit fracture site, and
immunohistochemical method was used to detect the protein expression of bFGF2 in fracture site of rabbits. RESULTS The healing
speed of the fracture site in the n-butanol fraction group was the fastest, followed by ethanol extract group, and the ethyl acetate
fraction group was the slowest; the serum levels of TNF-α and IL-6 in n-butanol fraction group decreased the fastest, while the
levels of ALP, bFGF2, OC and VEGFA increased the fastest [significant increase compared with ethanol extract group (P<0.01)];
the chondrocytes at the fracture fraction completely disappeared, forming a large number of bone marrow cavities, and the bone
trabeculae in the bone marrow cavity were officially formed. The expression level of bFGF2 was also higher than ethanol extract
group. CONCLUSIONS The effect of n-butanol fraction on promoting fracture healing is more significant than ethyl acetate
fraction and ethanol extract, and n-butanol fraction is the active fraction of Jiegu ointment to promote fracture healing.
KEYWORDS Jiegu ointment; fracture; healing; ethanol extract; n-butanol fraction; active fraction
现代医学认为,骨折愈合过程包含血肿炎症机化 化期内,骨折部位血管损伤形成血肿而分泌炎症因子
期、原始骨痂形成期、骨痂改造塑形期。在血肿炎症机 [如肿瘤坏死因子 α(tumor necrosis factor-α,TNF-α)和
白细胞介素 6(interleukin-6,IL-6)等]导致炎症反应,刺
Δ 基金项目 湖北省知识创新专项(自然科学基金)资助项目(No. 激受损部位,而具有消肿止痛功效的中药,可抑制炎症
2019CFB615) 因子过度表达而改善软组织损伤,并缩短骨折愈合的血
*第一作者 主治医师,硕士。研究方向:中药与康复医学。 肿炎症机化期 [1―3] 。在原始骨痂形成期内,血管形成先
E-mail:41629986@qq.com
于骨形成,为骨组织稳态维持提供坚实的物质基础,血
# 通信作者 主任医师,硕士生导师。研究方向:土家特色药材临
床研究与应用。E-mail:723677231@qq.com 管内皮生长因子 A(vascular endothelial growth factor A,
中国药房 2023年第34卷第11期 China Pharmacy 2023 Vol. 34 No. 11 · 1349 ·