Molecular mechanism of filiform needling and fire needling therapies in treatment of rat models of myelosuppression
Objective To explore the molecular mechanism of filiform needling and fire needling in the treatment of myelosuppressive rat models.Methods The study was from March to September 2022.Ninety-six SD rats who were 4-6 weeks old and whose body weight was 250-300 g were bought from Charles River.The rats were divided into a control group,a model group,a fire needle group,and a filiform needle group,with 24 in each group.Except the control group,the rats in the other groups were induced by cyclophosphamide(CTX)to establish the rat bone marrow suppression models.After the models were established successfully,the rats in the control group and the model group were grasped and fixed every day,but took no treatment.The rats in the fire needle group were fixed on the mouse boards in prone position according to the numbers,and took vertical acupuncture on their Zusanli points at 3 mm depth on both sides on the first,third,fifth,and seventh days after the modeling with the fire needles once every other day,for 7 consecutive days,and the needles were not left in the points.The rats in the filiform needle group were fixed on the mouse boards in prone position according to the numbers;the needles and acupoints were the same as those in the fire needle group;the needles were vertically punctured 3 mm into the points,and were left there for 6 min;the treatment started on the first day after modeling,once a day for 7 consecutive days.Ten μl blood was collected from the tail vein of the rats on the first to seventh days of intervention to measure the numbers of white and red blood cells and platelets,hemoglobin,and other blood routine indicators.The bone marrow tissue sections were made for for Wright's Giemsa staining.The flow cytometry was used to detect the bone marrow mononuclear fibroblast cycles.The expressions of cycle-related proteins in the bone marrow mononuclear fibroblasts were detected by Western blotting.LSD-t test,one-way analysis of variance,and Kruskal-Wallis test were applied.Results The numbers of white blood cells and platelets in the model group were lower than those in the control group(all P＜0.05),while the numbers of red blood cells and the contents of hemoglobin in the model group were higher than those in the control group(all P＜0.05).One,five,and seven days after the treatment,the numbers of white blood cells[(3.53±0.19)×109/L and(3.42± 0.22)×109/L vs.(3.12±0.10)×109/L;(3.78±0.23)×109/Land(3.65±0.29)×109/Lvs.(2.88±0.24)×109/L;(4.08±0.43)×109/L and(3.70±0.17)×109/L vs.(2.27±0.12)×109/L]and platelets[(79.72±3.89)×109/L and(79.15±5.22)×109/L vs.(70.17±2.20)×109/L;(85.18±3.40)×109/L and(81.88±4.97)×109/L vs.(62.73±3.80)×109/L;(92.18±7.64)×109/L and(83.45±4.29)×109/L vs.(51.92±1.64)×109/L]in the fire needle group and the filiform needle group were higher than those in the model group(all P＜0.05);the numbers of red blood cells[(1.36±0.03)× 1012/L and(1.37±0.03)×1012/L vs.(1.52±0.03)× 1012/L;(1.32±0.03)×1012/L and(1.34±0.02)×1012/L vs.(1.59±0.02)×1012/L;(1.24±0.07)×1012/L and(1.32±0.05)×1012/L vs.(1.68±0.03)×1012/L]and the contents of hemoglobin[(84.00±1.67)g/L and(84.33±1.86)g/L vs.(94.00±1.26)g/L;(82.00±2.68)g/L and(83.50±0.84)g/L vs.(97.83±1.17)g/L;(76.67±5.32)g/L and(81.83±3.43)g/L vs.(104.17±2.04)g/L]in the fire needle group and the filiform needle group were lower than those in the model group(all P＜0.05).The numbers of bone marrow blast cells in the filiform needle group and the fire needle group were higher than that in the model group,and the rod-shaped nuclei cells were found.The percentages of bone marrow mononuclear fibroblasts in the G0/G1 phase were lower than those of the model group(all P＜0.05);the change in the fire needle group was more significant than that in the filiform needle group,and the percentages of bone marrow mononuclear fibroblasts in the S phase and G2/M phase were always higher than those in the model group(all P＜0.05).The relative expressions of CD4,CD6,Cylcin D1,and E2F1 proteins in the rat bone marrow mononuclear fibroblasts were significantly up-regulated in the filiform needle group and fire needle group 3 and 7 d after the treatment(all P＜0.05).The expressions of cell cycle-related proteins,CD4,CD6,Cylcin D1,and E2F1,were more significantly up-regulated in the fire needle group.Conclusions Filiform needling and fire needling both can improve cyclophosphamide-induced myelosuppression in rats.The molecular mechanism may be related to up-regulating the expressions of cell cycle-related proteins,CD4,CD6,Cylcin D1,and E2F1,in rat bone marrow mononuclear fibroblasts,thereby restoring the metabolic balance of proliferation and dormancy in bone marrow stromal cells.
万方数据
维普
