| Purpose:Use guiluerxianjiao to treat GIOP zebrafish model.Observe its result and analyze how it can be worked.Methods:We used zebrafish and glucocorticoid dexamethasone to research the osteoporosis.We used HE and Masson staining to observe the microscopic structure of the zebrafish scales.Using Wb method to research zebrafish TGF-β,Smad3,P-Smad3 protein expression.Observations made mold results and analysis the relationship between OP and TGF-(3/Smad signaling pathways.Using the Guiluerxianjiao to the model,we can observe to the microscopic structure of the zebrafish scales By HE and Masson staining.Detection of zebrafish body inflammation factor IL-6,TNF-β,IL-1.Rt-pcr detection of zebrafish’s gene expression about TGF-β、Smad3、AP-1、NF-κBP65.Western blot method to detect zebrafish’s predominate protein expression about TGF-β、Smad3、P-Smad3、AP-1、P-NF-κBP65、NF-κBP65、P-IκBa、IκBa.Researching the mechanism of the treatment GIOP zebrafish model by the signal of TGF-β and NF-κB.Results:1,Successful establishment of GIOP zebrafish model,zebra fish scales by Masson staining and HE staining observation,blank group of zebrafish compact structure of type I collagen tissue,no apparent porosity.Compared with the blank group,dexamethasone low doseand medium dose groupwere no significant difference.Compared with the blank group,dexamethasone high dose set of disordered arrangement of type I collagen,collagen structure and fine,poor continuity,whichhas obvious difference.We use method of Western blot to research it’s related protein expression:compared with the blank group,dexamethasonelowdose and medium dose group were no significant difference,compared with the blank group,dexamethasone high dose set of TGF-β and Smad3 protein decreased obviously.2,By using Masson staining and HE staining,blank group of zebrafish compact structure of type I collagen tissue,no apparent porosity.Dexamethasone model group type I collagen arrangement happens disorder,slender collagen structure,poor continuity;Calcitriol and guiluerxianjiao’s low,medium and high dose group can significantly improve the above changes.3,By detecting zebrafish’s body inflammation factor indicators:compared with blank group,dexamethasone modelgroup’sIL-6、TNF-α、IL-1β content increased significantly;compared withdexamethasone modelgroup.Calcitriol and guiluerxianjiao’s low,medium and high dose group can significantly reduce the IL-6、TNF-αIL-1β content.4,By using method of rt-pcr to research it’s mRNA expression:compared with blank group,dexamethasonemodel group TGF-β and P-Smad3 content decreased significantly;however it’s NF-κBP65,AP-1 content increased;compared withdexamethasonemodel group,Calcitriol and guiluerxianjiao’s low,medium and high dose group of TGF-β and P-Smad3 content increased significantly;however it’s NF-κBP65,AP-1 content decreased.5,By using method of Western blot to research it’s protein expression:compared with blank group,dexamethasone model group TGF-β and P-Smad3 content decreased significantly,however it’s NF-κBP65、IκBα、AP-1 content increased significantly;compared with dexamethasone model group,Calcitriol and guiluerxianjiao’s low,medium and high dose group of TGF-β and P-Smad3 content increased significantly;however it’sNF-κBP65、IκBa、AP-1 content decreased.Conclusions:1.We have built zebrafish’s GIOP model successfully.2.Guiluerxianjiao can treated the OP through the TGF-β/Smad3 signaling pathway which can promotion of collagen.3.Guiluerxianjiao can treat OP through NF-κB and AP-1 which can relife the inflammation. |
