Abstract:

Objective　To explore the effect and possible mechanism of Yougui Pills assisting bone marrow mesenchymal stem cells (BMSCs) on healing of osteoporotic fractures in rabbits. Methods　This study was from January to June 2023. Fifty rabbits were divided into a normal control group, a model control group, a Yougui pill group, a BMSCs group, and a combined treatment group, with 10 in each group. Osteoporosis fracture models were established in all groups except the normal control group, and corresponding drugs were given. After the treatment, the bone mineral density (BMD), trabecular number, trabecular thickness, trabecular dispersion, elastic modulus, stiffness, maximum stress, and maximum bearing capacity of femur were measured. The levels of Janus kinase 2/signal transducer and activator of transcription 3 (JAK2/STAT3) gene and protein in femur were determined by real time polymerase chain reaction and Western blotting. One-way analysis of variance and LSD-t test were used to analyze the data. Results　The BMD, number of bone trabeculae, thickness of bone trabeculae, elastic modulus, stiffness, maximum stress, maximum bearing capacity, and levels of JAK2 and STAT3 mRNA and protein of femurs in the model control group were lower than those in the normal control group (P<0.05), and the trabecular dispersion in the model control group was higher than that in the normal control group (all P<0.05). The BMD, number of bone trabeculae, thickness of bone trabeculae, elastic modulus, stiffness, maximum stress, maximum bearing capacity, and levels of JAK2 and STAT3 mRNA and protein of femurs in the Yougui pill group, the BMSCs group, and the combined treatment group were higher than those in the model control group (all P<0.05). The trabecular bone dispersions in the Yougui pill group, the BMSCs group, and the combined treatment group were lower than that in the model control group (all P<0.05). The femoral BMD [(1.76±0.13) g/cm³], trabecular number [(5.87±0.24) pieces/mm], trabecular thickness [(0.14±0.03) mm], femoral elastic modulus [(343.27±21.04) MPa], stiffness [(904.25±73.25) N/mm], maximum stress [(32.63±3.17) N/mm²], maximum bearing capacity [(358.85±13.02) N], and levels of JAK2 and STAT3 mRNA [(3.45±0.26) and (3.60±0.28)] and protein [(0.82±0.05) and (0.84±0.05)] in the combined treatment group were higher than those in the Yougui pills group [(1.41±0.15) g/cm³, (3.88±0.28) pieces/mm, (0.10±0.02) mm, (304.62±20.41) MPa, (698.52±69.55) N/mm, (25.34±3.22) N/mm², (297.34±13.24) N, (2.58±0.26), (2.29±0.23), (0.60±0.05), and (0.53±0.07)] and the BMSCs group [(1.47±0.14) g/cm³, (3.89±0.28) pieces/mm, (0.09±0.03) mm, (299.54±16.94) MPa, (720.33±69.48) N/mm, (23.24±3.13) N/mm², (289.38±13.13) N, (2.63±0.25), (2.30±0.24), (0.59±0.06), and (0.53±0.08)] (all P<0.05); the bone trabecular dispersion [(0.34±0.02) mm] in the combined treatment group was lower than those in the Yougui pills group [(0.42±0.03) mm] and the BMSCs group [(0.43±0.03) mm] (both P<0.05). Conclusions　Yougui pills assisting BMSCs can significantly promote the healing of osteoporotic fractures in rabbits; the mechanism may be related to the activation of the JAK2/STAT3 signaling pathway of femur in rabbits with osteoporotic fractures.

Key words:

Osteoporosis fractures, Yougui Pills, Bone marrow mesenchymal stem cells, Healing, JAK2/STAT3 signaling pathway, Animal experiment

摘要：

目的　探讨右归丸辅助骨髓间充质干细胞对家兔骨质疏松性骨折愈合的影响及可能机制。方法　研究时间为2023年1月至6月。将50只家兔分为正常对照组、模型对照组、右归丸组、骨髓间充质干细胞组、联合治疗组，各10只。除正常对照组外其余各组建立骨质疏松性骨折模型，并予以相应药物治疗。治疗结束后，测定家兔股骨骨密度（bone mineral density，BMD）、骨小梁数量、骨小梁厚度、骨小梁分散度以及股骨弹性模量、刚度、最大应力、最大承载力；实时聚合酶链式反应及蛋白印迹法测定股骨Janus蛋白酪氨酸激酶2/信号转导和转录激活子3（Janus kinase 2/signal transducer and activator of transcription 3，JAK2/STAT3）基因与蛋白水平。采用单因素方差分析及LSD-t检验进行数据分析。结果　模型对照组家兔股骨BMD、骨小梁数量、骨小梁厚度，股骨弹性模量、刚度、最大应力、最大承载力，股骨JAK2、STAT3 mRNA和蛋白水平均低于正常对照组（均P<0.05），骨小梁分散度高于正常对照组（P<0.05）；右归丸组、骨髓间充质干细胞组、联合治疗组家兔股骨BMD、骨小梁数量、骨小梁厚度、股骨弹性模量、刚度、最大应力、最大承载力、股骨JAK2、STAT3 mRNA和蛋白水平均高于模型对照组（均P<0.05），骨小梁分散度低于模型对照组（均P<0.05）；联合治疗组股骨BMD［（1.76±0.13）g/cm³］、骨小梁数量［（5.87±0.24）个/mm］、骨小梁厚度［（0.14±0.03）mm］，股骨弹性模量［（343.27±21.04）MPa］、刚度［（904.25±73.25）N/mm］、最大应力［（32.63±3.17）N/mm²］、最大承载力［（358.85±13.02）N］，股骨JAK2、STAT3 mRNA［（3.45±0.26）、（3.60±0.28）］和蛋白［（0.82±0.05）、（0.84±0.05）］水平高于右归丸组［（1.41±0.15）g/cm³、（3.88±0.28）个/mm、（0.10±0.02）mm、（304.62±20.41）MPa、（698.52±69.55）N/mm、（25.34±3.22）N/mm²、（297.34±13.24）N、（2.58±0.26）、（2.29±0.23）、（0.60±0.05）、（0.53±0.07）］、骨髓间充质干细胞组［（1.47±0.14）g/cm³、（3.89±0.28）个/mm、（0.09±0.03）mm、（299.54±16.94）MPa、（720.33±69.48）N/mm、（23.24±3.13）N/mm²、（289.38±13.13）N、（2.63±0.25）、（2.30±0.24）、（0.59±0.06）、（0.53±0.08）］（均P<0.05），骨小梁分散度［（0.34±0.02）mm］低于右归丸组［（0.42±0.03）mm］、骨髓间充质干细胞组［（0.43±0.03）mm］（均P<0.05）。结论　右归丸辅助骨髓间充质干细胞能明显促进家兔骨质疏松性骨折愈合，其机制可能与右归丸辅助骨髓间充质干细胞治疗能激活骨质疏松性骨折家兔股骨JAK2/STAT3信号通路有关。

关键词:

骨质疏松性骨折, 右归丸, 骨髓间充质干细胞, 愈合, JAK2/STAT3信号通路, 动物实验