Study On Anti-senile Osteoporosis Mechanism Of Plastrum Testudinis Extract Based On NF-?B Signaling Pathway

Background:Senile Osteoporosis(SOP)is a systemic bone disease caused by a combination of age and other factors.As the population ages,the incidence of senile osteoporosis has increased year by year in the worldwide.Its high mortality and disability rate have become important health problems for human beings.So how to prevent and control SOP has also become a serious challenge for human being.Objective:The expression of nuclear factor-kappa B(NF-? B)in serum of patients with SOP was detected to investigate the changes of NF-? B in the pathogenesis of SOP.A SOP mice model was established and intervened by plastrum testudinis extract(PTE)or JSH-23(an inhibitor of NF-? B)to investigate the role of PTE in the regulation of NF-? B pathway in SOP prevention and treatment.Methods:1.Clinical ResearchForty serum samples from SOP patients or elderly patients without osteoporosis were collected to detect the content of NF-? B in serum.2.Animal research(1)Establishment and authentication of mice SOP model.Eighty 8-week-old female C57BL/J mice were divided into 4 groups,including sham operation group(SHAM),ovariectomized group(OP),senile group(S),and senile osteoporosis group(SOP),20 mice in each group.OP group and SOP group mice were ovariectomized bilaterally;SHAM group and S group were treated with the OP group surgical approach to remove small abdominal fat.After 1 week,S group and SOP group were subcutaneously injected with D-galactose solution every day.The general state and body of the mice were recorded in batches at week 4,week 8,and week 12.Then,in order to evaluated the aging and osteoporosis situation of mice,micro-CT was used to detect the microstructure of the lumbar vertebral body,morphology of the lumbar vertebrae and the structure of the hippocampal CA1 were observed by HE staining.(2)Treatment of SOP mice with PTE and the investigation of the possible mechanimsForty-four 8-week-old female C57BL/J mice were divided into four groups,including sham operation group(SHAM),senile osteoporosis group(SOP),NF-? B inhibition treatment group(INH),and PTE treatment group(PTE).SHAM group and S group were treated with a operation to remove small abdominal fat.The SOP group,the INH group,and the PTE group were treated by using the aforementioned method to establish SOP model.The INH group was administered with JSH-23 by gavage at the same time as D-galactose solution was injected,and the PTE gropu was given PTE by gavage while D-galactose solution was injected.The general state and body weight of the mice were recorded at 4th week and 8th week respectively.Micro-CT was used to detect the microstructure of the lumbar vertebral body,and morphology of the lumbar vertebrae and the structure of the hippocampal CA1 were observed by HE staining.qPCR was used to detect the expression levels of NF-K B,TNFR2 and ERa mRNA in vertebral bodies.Resuts:1.Clinical ResearchThe NF-? B expression level in the serum of patients with SOP was significantly higher than that in the control group(P<0.05).2.Animal research(1)Establishment and authentication of mice SOP model.?General situationThe mice in the SOP group were bald on the back,the surrounding hair was sparse and dull,the reaction was slow,the movement was slow,the lying was less,the diet was increased,and the right ear was dull.?Body weightAt the 4th week(W4),8 weeks(W8),and 12 weeks(W12)after modeling,the body weight of SOP group was higher than that of SHAM group,and there was statistical difference(P<0.05).?Microstructure of lumbar vertebraeAt W4,compared with SHAM mice,BV/TV was significantly decreased in SOP group(P<0.05),and SMI and Tb.Sp were significantly increased(P<0.05).At W8,compared with SHAM group,SMI and Tb.Sp in SOP group increased significantly(P<0.05),and BV/TV and Tb.Th decreased significantly(P<0.05).At W12,compared with SHAM group,SMI and Tb.Sp in SOP group increased significantly(P<0.05),and BV/TV,Tb.Th,Tb.N decreased significantly(P<0.05).?Morphological and structural changes in hippocampal CA1 regionAt W8 and W12,the neurons in the hippocampal CA1 region of the SOP group were disordered,the number of neuronal cells was significantly reduced,the neuronal cells were atrophied,and the intercellular space was significantly increased.(2)Treatment of SOP mice with PTE and the investigation of the possible mechanisms? The improvement of lumbar osteoporosis and hippocampal structure in each groupAt W8,compared with the SOP group,the SMI was decreased significantly,BV/TV was increased significantly(P<0.05),Tb.Th and Tb.N were increased slightly,and Tb.Sp was decreased slightly in PTE group.Compared with the SHAM group,the trabecular bone continuity of the INH group and the PTE group were worse.Compared with the SOP group,the trabecular bone was thickened and increased,and the trabecular gap was significantly narrowed in the visual field of the PTE group.Compared with the SOP group,the number of hippocampal neurons in the hippocampal CA1 region was increased,the cell arranged neatly,and the intercellular space was reduced.?Expression of NF-? B,TNFR2 and ERa mRNA in lumbar vertebrae of mice in each groupAt W8,compared with SOP group,the expression of NF-?B,TNFR2 and ERa in INH group and PTE group showed a downward trend,and the expression of ERa mRNA in PTE group was significantly down-regulated(P<0.05).Conclusion:1.Clinical ResearchNF-? B may be an important factor in the pathogenesis of SOP.2.Animal ResearchOvariectomy or D-galactose injection alone,and ovariectomy combined with D-galactose injection can effectively construct osteoporosis model,while ovariectomy combined with D-galactose injection can construct SOP model faster.NF-? B transcriptional activity may be a potential mechanism in the pathogenesis of SOP;PTE can effectively improve the micros tructure and bone morphology of mouse SOP model,and its mechanism may be related to the inhibition of NF-? B transcription.