Effect of Qing'e Formula on PI3K/Akt Signaling Pathway in Bone Tissue of Ovariectomized Osteoporosis Rats
Objective:To investigate the effect of Qing'e formula on the PI3K/Akt signaling pathway in bone tissue of ovariecto-mized osteoporosis rats.Methods:72 3-month-old SPF grade female SD rats were randomly divided into sham group,model group and Qing'e formula group using a random number table method,with 24 rats in each group.Rats were anesthetized by intraperitoneal injection of 2%pentobarbital sodium solution at a dose of 40 mg/kg,and an incision was made in the middle of the lower abdomen.Both the model and Qing'e formula groups had bilateral ovaries removed,while the sham group had a small amount of adipose tissue around the ovaries removed and immediately closed the abdomen and sutured the wound.After one week of modeling,the sham and model groups were given 2 mL of 0.9%sodium chloride solution by gavage,while the Qing'e formula group was given Qing'e for-mula solution by gavage at a dose of 0.42 g/(kg·d),once a day for 12 consecutive weeks.After intervention for 4,8,and 12 weeks,8 mice were euthanized and samples were taken from each of the 3 groups.The Masson staining method was used to observe the mor-phology of the femoral neck;femoral bone mineral density and maximum load of the femur were detected;ELISA method was used to detect serum PI3K and Akt level;qPCR and western blot were used to detect the mRNA relative expression level and protein expression of PI3K,Akt,and glycogen synthase kinase 3β(GSK3β)in the lumbar spine.Results:Compared with the sham group,the mRNA relative expression level of Akt in the model group significantly increased in the lumbar spine at week 4 of intervention(P<0.05),while the maximum load of the femur significantly decreased in the 12th week(P<0.05);at weeks 8 and 12,the femoral bone mineral density significantly decreased(P<0.05),and the mRNA relative expression level of PI3K in the lumbar spine signifi-cantly increased(P<0.05),while the protein expression of PI3K significantly increased(P<0.05);at weeks 4,8,and 12,the level of serum PI3K and Akt significantly increased(P<0.05),and the protein expression of Akt and GSK3β in the lumbar spine significantly increased(P<0.05),while the mRNA relative expression level of GSK3β significantly increased(P<0.05).Compared with the model group,the mRNA relative expression level and protein expression of Akt in the lumbar vertebrae of the Qing'e formula group significantly decreased at week 4(P<0.05);at the 12th week,the maximum load of the femur significantly increased(P<0.05),and the level of serum Akt significantly decreased(P<0.05),while the protein expression of Akt in the lumbar spine significantly decreased(P<0.05);at weeks 8 and 12,the femoral bone mineral density significantly increased(P<0.05),and the mRNA relative expression level of PI3K and GSK3β in the lumbar spine significantly decreased(P<0.05),while the protein expression level of PI3K signifi-cantly decreased(P<0.05);at weeks 4,8,and 12,the level of serum PI3K significantly decreased(P<0.05).Conclusion:Qing'e formula can improve bone mineral density and biomechanical properties in osteoporosis model rats,and its mechanism may be related to the inhibition of the PI3K/Akt signaling pathway.
osteoporosisovariectomized ratsQing'e formulabone mineral densitybiomechanicsPI3KAkt
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