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1.
Article in Chinese | WPRIM | ID: wpr-906511

ABSTRACT

Objective:To observe the effect of Zuoguiwan on bone metabolism and Wnt/<italic>β</italic>-catenin signaling pathway in ovariectomized osteoporotic rats model, and to explore the molecular biological mechanism of Zuoguiwan in the prevention and treatment of osteoporosis. Method:The rat model of postmenopausal osteoporosis was established by bilateral ovariectomy, 60 female SD rats were randomly divided into sham operation group, model group, positive group (estradiol valerate tablet 0.05 mg·kg<sup>-1</sup>·d<sup>-1</sup>) and low, middle and high dose groups of Zuoguiwan (5.5,11,22 g·kg<sup>-1</sup>·d<sup>-1</sup>).After successful establishment of the model in the 13<sup>th</sup> week, intragastric administration (<italic>ig</italic>) was given once a day for a total of 12 weeks. After administration, the histomorphological changes of femur in rats were observed by hematoxylin-eosin (HE) staining, the bone mineral density (BMD) and bone mineral content(BMC) of femur were measured by dual energy X-ray apparatus, and the biomechanical properties of bone were measured by MTS Acumen3 biomechanical testing system. The contents of bone alkaline phosphatase (BALP), bone glaprotein(BGP),estradiol (E<sub>2</sub>) ,and tartrate-resistant acid phosphatase (TRAP), type Ⅰ procollagen N-terminal propeptide (PINP) in serum were detected by enzyme-linked immunosorbent assay (ELISA). Western blot was used to detect the protein level of Wnt2,<italic>β</italic>-catenin,low density lipoprotein related receptor protein 5 (LRP5) and the phosphorylation level of glycogen synthase kinase-3<italic>β</italic>(GSK-3<italic>β</italic>) in rat tibia. Result:Compared with sham operation group, the maximum load and stiffness of BMD,BMC, in the model group decreased significantly(<italic>P</italic><0.01), the contents of E<sub>2</sub> and PINP in serum decreased significantly(<italic>P</italic><0.01), the content of BALP,BGP,TRAP increased significantly(<italic>P</italic><0.01), the expression levels of Wnt2,p-GSK-3<italic>β </italic>Ser9,LRP5 and <italic>β</italic>-catenin protein in bone tissue decreased significantly(<italic>P</italic><0.01), the trabecula of femur became thinner and thinner, the number of bone trabeculae decreased. Compared with model group, the maximum load and stiffness of BMD,BMC, in estradiol group and Zuoguiwan group were significantly increased (<italic>P</italic><0.05,<italic>P</italic><0.01), the contents of serum E<sub>2</sub> and PINP were significantly increased (<italic>P</italic><0.05,<italic>P</italic><0.01), the content of BALP,BGP,TRAP was significantly decreased (<italic>P</italic><0.01), and the expression level of Wnt2,p-GSK-3<italic>β</italic> Ser9,LRP5, <italic>β</italic>-catenin protein in bone tissue was significantly increased (<italic>P</italic><0.05,<italic>P</italic><0.01) , the trabeculae of femur became thicker, the number increased, the structure was basically clear. Conclusion:Zuoguiwan has a certain preventive and therapeutic effect on osteoporosis in ovariectomized rats, and its mechanism may be related to increasing the level of estrogen, activating Wnt/<italic>β</italic>-catenin signaling pathway, up-regulating the expression of Wnt2 and LRP5 protein, inhibiting the activity of GSK-3<italic>β</italic>, reducing the degradation of <italic>β</italic>-catenin, coordinating the dynamic coupling balance between bone formation and bone resorption, correcting the disorder of bone metabolism and improving bone morphology.

2.
Chinese Journal of Endemiology ; (6): 627-631, 2008.
Article in Chinese | WPRIM | ID: wpr-643005

ABSTRACT

Objective To investigate the protective mechanism of protein kinase C(PKC)and calcium sensing receptor(CaR)in ischemia preconditioned rat hearts.Methods Using cell culture method,in vitro cultured inhibitor(IPC+CaRI).Apoptosis was detected using TUNEL and Hoechst33342 cell viability was detected by MTT,the protein expression of easpase-12,calpain and CaR in endochylema were detected using Wedtetm blot.ResultsIn I/R group nucleus was shrank,big blue,chromatin concentrated,apoptotle body appeared.Other groups haddifferent fluorescence intensity varying degree,IPC+PKCI+CaRS group had more big blue nucleus.Myocardialcell viability and apoptotic rate,I/R group[(62.99±0.65)%,(19.13±0.87)%],IPC group[(78.67±0.37)%,(14.21±0.74)%],IPC+PKCI group[(71.09±0.52)%,(20.46±0.81)%],IPC+PKCI+CaRS group(66.10±0.75)%,(24.89±1.43)%],IPC+CaRS group[(69.56±0.44)%,(21.64±0.77)%],IPC+CaRI group(85.81±0.60)%,(13.12±0.69)%],all had a difference(P<0.05 or<0.01)compared with C group[(100.00)%,(6.02±0. 31)%].Western blot identified that CaR expression in IPC+PKCI and IPC+CaRS,IPC+PKCI+CaRS groupswas more than that in IPC and IPC+CaRI groups;easpase-12 had more active fragment(60×103)in I/R,IPC+CaRS,IPC+PKCI+CaRS groups;ealpain expressions in I/R,IPC,IPC+PKCI,IPC+PKCI+CaRS,IPC+CaRSgroups were higher than those in C and IPC+CaRI,I/R group was the highest one,C group the second,IPC+CaRI the third.Conclusion The interaction of PKC and CaR can reduce the intracellular Ca2+ from sarcoplasmicreticulum thus provide a protection.

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