[1]宋飞,江敏,李世元,等.藏红花素水凝胶干预大鼠胫骨骨折的疗效及作用机制研究[J].中医正骨,2025,37(10):23-29，37.
　SONG Fei,JIANG Min,LI Shiyuan,et al.Efficacy and mechanism of crocin hydrogel against tibial fractures in rats:an experimental study[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2025,37(10):23-29，37.
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藏红花素水凝胶干预大鼠胫骨骨折的疗效及作用机制研究()

《中医正骨》[ISSN:1001-6015/CN:41-1162/R]

卷:
第37卷
期数:
2025年10期
页码:
23-29，37
栏目:
基础研究
出版日期:
2025-10-20

文章信息/Info

Title:
Efficacy and mechanism of crocin hydrogel against tibial fractures in rats:an experimental study
作者:
宋飞; 江敏; 李世元; 陈前永
中国人民解放军联勤保障部队第九〇一医院,安徽 合肥 230031
Author(s):
SONG Fei; JIANG Min; LI Shiyuan; CHEN Qianyong
The 901st Hospital of the Joint Logistics Support Force of the Chinese People's Liberation Army,Hefei 230031,Anhui,China
关键词:
胫骨骨折;  骨折愈合;  藏红花素;  水凝胶类;  大鼠;  动物实验
Keywords:
tibial fractures;  fracture healing;  crocin;  hydrogels;  rats;  animal experimentation
摘要:
目的:探讨藏红花素水凝胶干预大鼠胫骨骨折的疗效及作用机制。方法:将30只雄性SD大鼠随机分为正常对照组、空白水凝胶组和藏红花素水凝胶组,每组10只。空白水凝胶组和藏红花素水凝胶组大鼠通过手术建立左侧胫骨骨折模型,在缝合切口前,在空白水凝胶组大鼠骨折断端注射0.2 mL空白水凝胶,在藏红花素水凝胶组大鼠骨折断端注射0.2 mL藏红花素水凝胶。正常对照组大鼠不进行手术。造模和干预后4周,采集腹主动脉血,检测大鼠血清碱性磷酸酶(alkaline phosphatase,ALP)、Ⅰ 型前胶原氨基端前肽(N-terminal propeptide of typeⅠprecollagen,PⅠNP)、骨钙素水平; 截取大鼠左侧胫骨组织(空白水凝胶组和藏红花素水凝胶组大鼠截取骨痂上下约0.5 cm的胫骨组织,正常对照组大鼠截取胫骨结节下方长约1 cm的胫骨组织),采用Micro-CT分析胫骨组织骨体积分数、骨小梁分离度、骨小梁数量、骨小梁厚度,采用HE染色观察胫骨骨痂组织病理改变,采用实时定量PCR检测胫骨组织骨形态发生蛋白(bone morphogenetic protein,BMP)2、Runt相关转录因子2(Runt-related transcription factor 2,RUNX2)的mRNA表达水平,采用Western Blot法检测胫骨组织BMP2、RUNX2、磷脂酰肌醇3激酶(phosphoinositide 3-kinase,PI3K)、蛋白激酶B(protein kinase B,PKB)、磷酸化PI3K、磷酸化PKB的蛋白表达水平,并计算PI3K和PKB的磷酸化水平。结果:①血清骨形成标志物检测结果。空白水凝胶组大鼠血清ALP、PⅠNP、骨钙素水平均高于正常对照组(P=0.000,P=0.000,P=0.000),藏红花素水凝胶组大鼠血清ALP、PⅠNP、骨钙素水平均高于空白水凝胶组(P=0.000,P=0.000,P=0.000)。②胫骨组织骨微结构检测结果。空白水凝胶组大鼠胫骨组织骨体积分数、骨小梁厚度均高于正常对照组(P=0.000,P=0.000),骨小梁分离度低于正常对照组(P=0.000),骨小梁数量多于正常对照组(P=0.000); 藏红花素水凝胶组大鼠胫骨组织骨体积分数、骨小梁厚度均高于空白水凝胶组(P=0.000,P=0.000),骨小梁分离度低于空白水凝胶组(P=0.000),骨小梁数量多于空白水凝胶组(P=0.000)。③胫骨骨痂组织病理学观察结果。空白水凝胶组大鼠的胫骨骨痂组织可见少量排列不规则的纤维组织,藏红花素水凝胶组大鼠的胫骨骨痂组织可见大量排列规则的纤维组织,且部分纤维组织已被正常骨组织替代。④胫骨组织成骨细胞分化相关基因的mRNA和蛋白表达水平检测结果。空白水凝胶组大鼠胫骨组织BMP2、RUNX2的mRNA和蛋白表达水平均高于正常对照组(P=0.000,P=0.000; P=0.000,P=0.000),藏红花素水凝胶组大鼠胫骨组织BMP2、RUNX2的mRNA和蛋白表达水平均高于空白水凝胶组(P=0.000,P=0.000; P=0.000,P=0.000)。⑤胫骨组织PI3K-PKB信号通路关键蛋白磷酸化水平检测结果。空白水凝胶组大鼠胫骨组织PI3K和PKB的磷酸化水平均高于正常对照组(P=0.000,P=0.000); 藏红花素水凝胶组大鼠胫骨组织PI3K和PKB的磷酸化水平均高于空白水凝胶组(P=0.000,P=0.000)。结论:藏红花素水凝胶干预大鼠胫骨骨折,能够改善胫骨骨折端骨微结构、促进骨折愈合,其作用机制可能与激活PI3K-PKB信号通路、促进成骨细胞分化相关基因表达有关。
Abstract:
Objective:To investigate the therapeutic efficacy of crocin hydrogel against tibial fractures in rats,and to explore its underlying mechanism.Methods:Thirty male SD rats were selected and randomized into normal control group,blank hydrogel group,and crocin hydrogel group,with 10 cases in each group.The rats in blank hydrogel group and crocin hydrogel group were subjected to surgeries on the left lower limbs to build tibial fracture models.Before suturing the incision,the blank hydrogel(0.2 mL)and crocin hydrogel(0.2 mL)were injected into the fractured sites of rats in the corresponding group,respectively,while the ones in normal control group were not given any surgical procedure.Four weeks after successful modeling and intervention,the blood was drawn from the abdominal aorta of rats in each group to measure the serum levels of alkaline phosphatase(ALP),N-terminal propeptide of typeⅠprecollagen(PⅠNP),and osteocalcin(OC).In addition,the left tibial tissues(approximately 0.5 cm above and below the callus in blank hydrogel group and crocin hydrogel group,and about 1 cm segment below the tibial tuberosity in normal control group)were harvested from the rats in each group to evaluate the bone volume/total volume(BV/TV),trabecular separation(Tb.Sp),trabecular number(Tb.N),and trabecular thickness(Tb.Th)in the tibial tissues by micro-CT,and examine the histopathological changes in the tibial callus by HE staining.Furthermore,the mRNA expression levels of bone morphogenetic protein 2(BMP2)and Runt-related transcription factor 2(RUNX2)in tibial tissues were detected by real-time quantitative PCR(RT-qPCR),and the protein expression levels of BMP2,RUNX2,phosphoinositide 3-kinase(PI3K),protein kinase B(PKB),phosphorylated PI3K(p-PI3K),and phosphorylated PKB(p-PKB)were measured by Western Blot,and the phosphorylation levels of PI3K and PKB were then calculated.Results:①Serum bone formation markers.The serum levels of ALP,PⅠNP,and OC were higher in blank hydrogel group compared to normal control group(P=0.000,P=0.000,P=0.000),and these markers were further elevated in crocin hydrogel group compared to blank hydrogel group(P=0.000,P=0.000,P=0.000).②Bone microstructure of tibial tissues.The BV/TV and Tb.Th in the tibial tissues were higher,the Tb.Sp was lower,and the Tb.N was more in blank hydrogel group compared to normal control group(P=0.000,P=0.000,P=0.000,P=0.000),and these parameters were further improved in crocin hydrogel group compared to blank hydrogel group(P=0.000,P=0.000,P=0.000,P=0.000).③Histopathological findings in tibial callus tissues.The changes,manifesting as a small amount of irregularly arranged fibrous tissues in the tibial callus tissues,were observed in rats from blank hydrogel group,whereas the abundant regularly arranged fibrous tissues was present in the tibial callus tissues of rats from crocin hydrogel group,with partial areas already replaced by the normal bone tissues.④The mRNA and protein expression levels of osteoblast differentiation-related genes in the tibial tissues.The mRNA and protein expression levels of BMP2 and RUNX2 in the tibial tissues were higher in blank hydrogel group compared to normal control group(P=0.000,P=0.000; P=0.000,P=0.000),and these markers were further upregulated in crocin hydrogel group compared to blank hydrogel group(P=0.000,P=0.000; P=0.000,P=0.000).⑤Phosphorylation levels of key proteins in the PI3K-PKB signaling pathway in the tibial tissues.The phosphorylation levels of PI3K and PKB in the tibial tissues were higher in blank hydrogel group compared to normal control group(P=0.000,P=0.000),and they were further increased in crocin hydrogel group compared to blank hydrogel group(P=0.000,P=0.000).Conclusion:Crocin hydrogel can improve the bone microstructure of the fractured site and promote the fracture healing in intervention of tibial fracture in rats.It may exert the effects by activating PI3K-PKB signaling pathway and upregulating the expression of osteoblast differentiation-related genes.

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通讯作者:宋飞 E-mail:13721039468@163.com
更新日期/Last Update: 1900-01-01