| Objective:The pathway and key targets of tortoise shell gum intervention in postmenopausal osteoporosis were analyzed by network pharmacology and molecular docking technology,and experimentally verified,which opened up new ideas for the treatment of postmenopausal osteoporosis and the research and development of new drugs.Methods:The main components of tortoiseshell gum were excavated through literature data,and the corresponding targets of the components were backscreened from Swiss target Prediction,and then the differential expression analysis of disease targets was carried out through GEO database to obtain differential expression targets of postmenopausal osteoporosis,and the target of tortoiseshell gum for intervention in postmenopausal osteoporosis was obtained after intersection,and the protein interaction network of intersecting genes was obtained through String 10.0.AutoDock software was used to carry out molecular docking of the intersection target and its corresponding components to observe whether its binding was stable.The targets obtained were verified by animal experiments,and 40 adult female SD rat animals were weighed and stratified after 1 week of rearing,and divided into 6 groups by random number table method,in order:model group,sham surgery group,estrogen group,tortoise shell gum low-dose group,tortoiseshell gum medium-dose group,and tortoiseshell gum high-dose group.The daily gavage of tortoiseshell gum was 0.63g/kg/d,1.26g/kg/d,and 2.52g/kg/d per day after two months of molding.The estrogen group was gavaged 0.009mg/mL daily after two months of modeling,and the model group and the sham group were given an equal volume of normal saline daily gavage after two months of modeling.Gastric gavage lasted a total of three months in each group.Groups of rats were sacrificed three months later.After sacrifice,bone tissue morphology was observed by bone tissue section analysis of rats.The expression of relevant target veneers obtained by network pharmacological analysis was determined by Western Blot method by gene protein expression detection,and finally statistical analysis was carried out after the data.Results:1.Through network pharmacology and molecular docking technology,it is concluded that tortoiseshell gum can intervene in postmenopausal osteoporosis through multiple targets and pathways,which has certain therapeutic significance,and is likely to play a key role in core targets such as SRC,ITGB1,and PIK3CA.Key targets can be enriched in the immune system,receptor tyrosine kinase signaling,interleukin signaling,PI3K/AKT signaling and other signaling pathways.Target ITGB1 is most likely to interact with N-Carbobenzoxy-Dl-Leucine.2.Bone tissue section:HE staining of the proximal femur tissue of the right femur showed that the femoral tissue structure of the rats in the sham surgery group was basically normal.Modular rats can see thin and loose trabeculae.The bone trabecular of the low-dose tortoiseshell gum group is thin and loose.The cartilage surface of the intoiseshell gum and the high dose group were smooth and complete,and no fragmentation and shedding were seen.The cartilage surface of the estrogen group was smooth and complete,and no fragmentation and shedding were seen.3.Detection of key gene protein expression:Compared with the sham group,the expression of SRC and ITGB1 protein was significantly increased in the model group,and the expression of PIK3CA protein was significantly reduced(P<0.01).There were no significant differences in the expression of SRC,ITGB1 and PIK3CA proteins in the middle and high dose groups of tortoiseshell gum and the estrogen group(P>0.05).Compared with the model group,the expression of SRC and ITGB1 in the dose of tortoiseshell gum was significantly reduced,and the expression of PIK3 C A protein was significantly increased(P<0.01).Conclusion:1.Through the results of network pharmacology and molecular docking,the targets of tortoiseshell gum in postmenopausal osteoporosis may be SRC,ITGB1,PIK3CA,etc.,which can play a multi-component and multi-pathway role in intervening postmenopausal osteoporosis rats.2.The ways of tortoi seshell gum to intervene in postmenopausal osteoporosis rats include the immune system,receptor tyrosine kinase signaling,interleukinin signaling,PI3K/AKT signaling and other signaling pathways.The target ITGB1 is most likely to interact with N-Carbobenzoxy-D1-Leucine in tortoiseshell gum.It opens up a new direction for the drug research of postmenopausal osteoporosis,and also provides a certain reference for the research and development of new drugs.3.An appropriate concentration of tortoiseshell gum can change the bone tissue morphology of postmenopausal rats and achieve the effect of alleviating postmenopausal osteoporosis in rats.4.An appropriate concentration of tortoiseshell gum can intervene in postmenopausal osteoporosis rats by reducing the expression of SRC and ITGB1 and increasing the expression of PIK3CA. |
PDF Full Text Request