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AIM:To investigate the effect of Bushen formulae(BHF)on bone metabolism and its possible mechanism in ovariectomized rats with high salt intake.METHODS:According to the random number table method,80 SPF-grade Sprague-Dawley rats were divided into sham group,ovariectomy(OVX)group,medium-high-salt diet(MSD)group,high-salt diet(HSD)group,BHF group,BHF with normal saline(BHF+NS)group,BHF+MSD group,and BHF+ HSD group,with 10 rats in each group.After modeling,different diets and BHF formula interventions were administered,and the concentrations of sodium chloride added to MSD group and HSD group were 2%(w/w)and 8%(w/w),respective-ly.The dose of BHF was 7.8 g·kg-1·d-1,once a day,and the treatment lasted for 12 weeks.Bone density,bone microar-chitecture,bone parameters,bone metabolism biomarkers,bone histopathological changes,the expression of epithelial sodium channel α(ENaCα),Na-Cl cotransporter(NCC),and voltage-gated chloride channel 3(ClC-3)proteins in bone tissue were detected in each group.RESULTS:Compared with sham group,the rats in OVX group had reduced bone density and destroyed bone microstructure.Compared with OVX group,the bone microstructure in MSD and HSD groups was more significantly damaged,while the levels of bone formation markers,bone glycoprotein(BGP)and type Ⅰ procolla-gen N-terminal peptide(PINP),were significantly increased in HSD group(P<0.05).Moreover,the levels of bone re-sorption markers,such as amino-terminal cross-linked telopeptides of type Ⅰ collagen(NTX),carboxy-terminal cross-linked telopeptides of type Ⅰ collagen(CTX)and tartrate-resistant acid phosphatase(TRACP),were significantly in-creased(P<0.05),indicating that bone metabolism was in high-conversion state.High-salt diet accelerated the structural destruction of bone trabeculae,and Western blot results showed that high-salt diet caused decreases in the protein expres-sion levels of ENaCα and ClC-3 and an increase in the protein expression level of NCC in femoral tissues(P<0.05).After BHF intervention,the expression of relevant ion channels caused by high salt could be regulated to different degrees.CONCLUSION:Bushen formulae could differentially regulate the expression of relevant ion channels ENaCα,ClC-3,and NCC induced by high salt to different degrees,which has certain ameliorative and therapeutic effects on the imbalance of bone metabolism.
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Objective To investigate the effect of chaihu-shugan-san ( CSS) on the behaviors and Notch1 signal pathway in depression model rats. Methods Thirty-two SD rats with similar behavioral scores were divided into control group (CON),model group (CUMS),positive control group (FLU) and interven-tion group (CSS).The depression model was established by stimulating with chronic unpredictable mild stress (CUMS),and the behaviors evaluation was assessed by sugar water consumption and forced depression.Im-munofluorescence was used to detect the proliferation of hippocampus neurons in rats,at the same time,real- time PCR and Western blot were used to detect the mRNA and protein expression levels of each factor (Notch1,Hes1,Hes5 and Jagged1) of Notch1 signal pathway respectively. Results Compared with CON group,the percentage of sugar water preference and swimming length of rats decreased significantly in CUMS group (P＜0.05 and P＜0.01).Compared with CUMS group,the percentage of sugar water preference and swimming length of rats increased significantly in CSS group(P＜0.05).Compared with CON group,there was a significant increase in the inactivity length of rats between CUMS group,FLU group and CSS group,and the difference was statistically significant (P＜0.01).Compared with CUMS group,the swimming length of rats in CSS group was significantly reduced,and the difference was statistically significant (P＜0.01).Compared with CON group((750.00±27.51)/mm2),the number of BrdU positive cells in the substratum or granulocyte lay-er of the hippocampus dentate gyrus of rats in CUMS group ((338.75±29.61)/mm2),FLU group ((545.00 ±17.73)/mm2) and CSS group ((529.38±13.74)/mm2) was significantly reduced(P＜0.01).Compared with CUMS group ((338.75±29.61)/mm2),there was a significant increase in the number of BrdU positive cells in the substratum or granulocyte layer of the hippocampus dentate gyrus of rats in CSS group ((529.38 ±13.74)/mm2),and the difference was statistically significant (P＜0.01).Compared with CON group,the mRNA and protein expression levels of Notch1,Hes1,Hes5,and Jagged1 in the hippocampus of rats in CUMS group were significantly reduced(P＜0.05 or P＜0.01).Compared with CUMS group,the mRNA and protein ex-pression levels of Notch1,Hes1,Hes5 and Jagged1 in the hippocampus of rats in FLU group and CSS group were significantly increased,and the differences were statistically significant (P＜0.05 or P＜0.01). Conclu-sion Notch1 signal pathway may be related to the obstacle during the hippocampus nerve regenerating in the model rat under chronic unpredictable mild stress.CSS may play an anti-depressant role by regulating Notchl to improve hippocampus nerve regeneration.