| BackgroundOsteoporosis(osteoporosis,OP)is a systemic bone disease characterized by decreased bone mass and deterioration of bone tissue microstructure,resulting in increased bone brittleness and fracture susceptibility.The formation mechanism of OP is that the activity of osteoclasts is higher than that of osteoblasts,resulting in greater bone resorption than bone formation,resulting in the decrease of bone mass,the destruction of bone microstructure and the decrease of biomechanical properties of bone tissue,which is easy to cause fracture.Osteoporosis is divided into primary and secondary osteoporosis.Primary osteoporosis includes postmenopausal osteoporosis,senile osteoporosis and idiopathic osteoporosis.Osteoporosis can occur at any age,but it is more common in elderly men and postmenopausal women.With the rapid increase of the average age of the world population,especially in developing countries,the prevalence of osteoporosis has also increased significantly,which has become a major public health problem.ObjectiveThe purpose of this study was to investigate the effect of zoledronic acid injection combined with Tenghuang Jiangu capsule on osteoporosis in ovariectomized SD rats by observing the bone turnover indexes(N-terminal propeptide of type I collagen,osteocalcin,alkaline phosphatase,C-terminal peptide of type I collagen,tartrate resistant acid phosphatase,etc.).To provide a theoretical basis for the clinical treatment of osteoporosis patients with the combination of traditional Chinese and western medicine in the future.Methods1.60 healthy female SD rats weighing 220 g were randomly divided into sham operation group(n=15)and ovariectomized group(n=45),postmenopausal osteoporosis model was established in ovariectomized group,and only bilateral periovarian fat was removed in sham operation group.12 weeks later,5 rats were killed in sham operation group and ovariectomized group,and the bone morphology of femurs of rats in each group was observed by hematoxylin-eosin staining.2.After verifying the successful establishment of osteoporosis model,the remaining 40 rats in the ovariectomized group were randomly divided into four groups: ovariectomized control group(n=10),zoledronic acid group(n=10),Tenghuang Jiangu group(n=10)and combined drug group(n=10).Zoledronic acid group was given intraperitoneal injection of zoledronic acid 0.5mg/kg,only once.The rats in the Tenghuang Jiangu group were given Teng Huang Jiangu by intragastric administration,0.18g/kg,once a day.The rats in the combination group were given intraperitoneal injection of zoledronic acid 0.5mg/kg,only once,and Teng Huang Jiangu was given intragastric administration,0.18g/kg,once a day.The sham operation group and the ovariectomized control group were given the same amount of normal saline once a day.During the period of intragastric administration,water was eaten freely and weighed once a week to adjust the dose for12 weeks.After 12 weeks,the indexes of bone turnover(N-terminal propeptide of type I collagen,osteocalcin,alkaline phosphatase,C-terminal peptide of type I collagen,tartrate-resistant acid phosphatase)in the serum of rats in each group were examined.The bone morphology of femurs of rats in each group was observed by hematoxylin-eosin staining.3.The data were analyzed by SPSS26.0 software.The measurement data were tested by normality and homogeneity of variance,which accorded with normal distribution by mean ±standard deviation(Mean±SD).The comparison between groups was made by single factor analysis of variance(p < 0.05),indicating that the difference was statistically significant.Results1.The osteoporosis model was successfully established: the bone morphology of femur specimens of rats in each group was observed by hematoxylin-eosin staining.The results showed that the femoral trabeculae of rats in the ovariectomized group were degenerative,sparse and broken,and the chondrocytes decreased or even disappeared.necrotic bone fragments were formed and adipocytes increased.2.The serum N-terminal propeptide of type I collagen in the sham operation group was significantly lower than that in the ovariectomized control group(p < 0.05),and that in the ovariectomized control group was significantly higher than that in the zoledronic acid group and the combination group(p < 0.05).The level in zoledronic acid group was significantly lower than that in Teng Huang Jiangu group and combination group(p < 0.05),and that in Teng Huang Jiangu group was significantly higher than that in combination group(p < 0.05).The serum osteocalcin in the sham operation group was significantly lower than that in the ovariectomized control group(p <0.05),and that in the ovariectomized control group was significantly higher than that in the zoledronic acid group,Teng Huang Jiangu group and the combination group(p < 0.05).The serum osteocalcin in the zoledronic acid group was significantly lower than that in the Teng Huang Jiangu group(p < 0.05).There was no significant difference compared with the combination group(p > 0.05),but it was significantly higher in the Teng Huang Jiangu group than in the combination group(p < 0.05).The level of serum alkaline phosphatase in sham operation group was significantly lower than that in ovariectomized control group(p <0.05),and that in ovariectomized control group was significantly higher than that in zoledronic acid group,Tenghuang Jiangu group and combination group(p < 0.05).The level of serum alkaline phosphatase in zoledronic acid group was significantly lower than that in Tenghuang Jiangu group and combination group(p < 0.05),but there was no significant difference between Teng Huang Jiangu group and combination group(p > 0.05).The serum C-terminal peptide of type I collagen in the sham operation group was significantly lower than that in the ovariectomized control group(p < 0.05),and that in the ovariectomized control group was significantly higher than that in the zoledronic acid group,Teng Huang Jiangu group and the combination group(p < 0.05).The level in the zoledronic acid group was significantly lower than that in the Teng Huang Jiangu group(p < 0.05),but there was no statistical difference compared with the combined group,but it was significantly higher in the Teng Huang Jiangu group than in the combined group(p < 0.05).The level of serum tartrate-resistant acid phosphatase in sham operation group was significantly lower than that in ovariectomized control group(p < 0.05),and that in ovariectomized control group was significantly higher than that in zoledronic acid group,Teng Huang Jiangu group and combination group(p < 0.05).The level in zoledronic acid group was significantly lower than that in Tenghuang Jiangu group and combination group(p < 0.05),and that in Tenghuang Jiangu group was significantly higher than that in combination group(p < 0.05).3.HE staining method was used to observe the bone morphology of rat femur samples in each group:In the sham operation group,the thickness of cartilage of the femoral head was consistent,no calcified chondrocytes were found,the bone trabeculae were dense and thick,the bone trabeculae were broken less,the osteocytes were normal and evenly distributed,and no necrotic bone was found.Ovariectomized control group: degeneration occurred in the process of osteogenesis,calcium salt deposition,bone trabecular degeneration,bone trabeculae significantly decreased,broken,chondrocytes decreased or even disappeared;necrotic bone fragments were formed;dense bone gradually degenerated and could not regenerate bone.Zoledronic acid group: there was calcification in the metaphysis,but the chondrocytes had been formed,the bone trabeculae had not been fully formed,and the old bone trabeculae had been restored,but the bone trabeculae could not be rebuilt.the new chondrocytes gradually formed bone,and the calcium deposition in compact bone gradually decreased and improved.Tenghuang Jiangu group: normal cartilage osteogenic matrix recovered,but not perfect,new chondrocytes appeared,calcium deposition basically disappeared,chondrocytes formed osteoblast process appeared,osteogenic process resumed,bone trabeculae were established imperfectly,necrotic bone disappeared,bone trabeculae began to proliferate from the edge,osteoblasts increased,density increased,new bone trabeculae formed,dense bone gradually recovered and calcification disappeared,but the dense bone was still loose.Not yet fully recovered,we can see the process of chondrocytes forming osteoblasts and new bone.In the combined treatment group,the degeneration and calcification of chondrocytes basically disappeared,there was almost no calcium deposition,new bone trabeculae formed,chondrocytes formed osteoblasts,further formed osteocytes,and the process of forming bone trabeculae appeared.the new bone trabeculae gradually extended to the bone marrow and the dense bone calcification disappeared.Conclusion1.The removal of bilateral ovaries in rats can increase the bone turnover index and the bone trabecular degeneration,decrease and necrotic bone formation,and then lead to osteoporosis.2.Zoledronic acid injection,Tenghuang Jiangu capsule and their combination can effectively improve the condition of osteoporosis in ovariectomized rats,but Tenghuang Jiangu may or only weakly inhibit the function of osteoclasts,its main effect may be to stimulate osteoblast activity and promote bone formation,while zoledronic acid mainly inhibits bone resorption. |
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