ABSTRACT
Objective:
To observe the repair effect and possible mechanism of
Dipsacus saponins Ⅵ on
tibial fracture model
rats.
Methods:
Thirty Sprague Dawley (SD)
rats were randomly divided into model group, intervention group, and combination group, with 10
rats in each group, to establish a
tibial fracture rat model using the sawing
method. On the second day after
surgery, the intervention group was intraperitoneally injected with 10 mg/kg of Chuanduduan
saponin Ⅵ; The combination group received
intraperitoneal injection of
Dipsacus saponins Ⅵ 10 mg/kg and XAV939 1 mg/
animal; The model group was intraperitoneally injected with 0.2 ml of physiological
saline solution and 0.2 ml of
dimethylsulfoxide (
DMSO)
solution; Once a day, continuous intervention for 14 days. After 2 to 4 weeks of intervention, Micro CT scan and
X-ray scan were used to observe the
fracture healing status; After 4 weeks of intervention, the wet weight of the
tibia was detected;
Hematoxylin eosin (HE)
staining was used to observe the pathological changes of
callus tissue; The
Western blot method was used to detect the expression level of
callus tissue β-
catenin (β-
catenin), p-β-
catenin,
glycogen synthase kinase 3β (
GSK-3 β) and Runt related
transcription factor 2 (Runx2)
protein.
Results:
After 2 and 4 weeks of intervention, the
bone volume fraction (BV/TV), number of trabeculae (Tb.N), Lane Sandhu score, and
callus volume in the intervention group were higher than those in the model group (all P<0.05); After 2 and 4 weeks of intervention, the BV/TV, Tb.N, Lane Sandhu score, and
callus volume in the combined group were lower than those in the intervention group (all P<0.05). The wet weight of the
tibia in the intervention group was higher than that in the model group at 4 weeks after intervention ( P<0.05); The wet weight of the
tibia in the combined group was lower than that in the intervention group ( P<0.05). The HE
staining results showed that the model group had fibrous
tissue hyperplasia and more
bone trabeculae, but the maturity was not high and the thickening was not significant; The intervention group formed more
bony callus, with orderly arrangement of
bone trabeculae, partially mature, and obvious mineralization, consistent with the direction of stress; The combined group formed more cartilaginous and fibrous
callus, with more mineralization at the edge of the cartilaginous
callus and the formation of
bone trabeculae. Abundant
capillaries can be observed in the gaps. The expression level of Runx2 and p-β-
catenin/β-
catenin protein in
callus tissue of the intervention group was higher than that of the model group, the
protein expression
GSK-3 β level was lower than that of the model group (all P<0.05); The expression level of Runx2 and p-β-
catenin/β-
catenin protein in the
callus tissue of the combined group was lower than that of the intervention group; the
protein expression level of
GSK-3β was higher than that of the intervention group (all P<0.05).
Conclusions:
Dipsacus saponins Ⅵ can effectively promote fracture repair in
tibial fracture model
rats; It is possible to
plays a
role by activating the Wnt/β-
catenin signaling pathway.