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OBJECTIVE:To study the time-effect and dose-effect of paeonol on the apoptosis of knee osteoarthritis(OA)artic-ular chondrocyte in rabbits and the mRNA expression of its related protein Bcl-2 and Bax. METHODS:60 big-ear rabbits were ran-domly divided into normal (normal saline) group,model (normal saline) group,paeonol high-dose,medium-dose and low-dose groups and triamcinolone acetonide(positive drug)group,with 10 rabbits in each group. Except for normal group,OA model was induced by right knee anterior cruciate ligament (ACLT) and the medial meniscus 1/3 resection in those groups. After modeling, different doses of paeonol(0.8,0.4,0.2 mg/kg),triamcinolone acetonide 0.2 mg/kg were injected into right articular cavity twice a week. 4 weeks and 8 weeks after administration,articular cartilage specimens were collected. Ultrapathological structure changes of articular chondrocytes were observed by electron microscope. Apoptosis of cartilage cell was observed by TUNEL and apoptotic index was calculated. mRNA expression of apoptosis related genes of Bcl-2 and Bax in articular cartilage tissue of rabbits were de-tected by in situ hybridization technique. RESULTS:Compared with normal group,articular chondrocyte of model group showed early and middle stage apoptosis morphology change after 4 and 8 weeks,and apoptosis index increased significantly and the mRNA expression of Bcl-2 and Bax was up-regulated (P<0.01);4 and 8 weeks later after administration,compared with model group,apoptosis index decreased and mRNA expression of Bax was down-regulated in paeonol groups,while mRNA expression of Bcl-2 was up-regulated(P<0.05 or P<0.01). Electron microscopy ultrastructural observation showed articular chondrocyte of pae-onol high-dose and middle-dose groups were in early stage of apoptosis.CONCLUSIONS:Paeonol can slow down articular chondro-cyte degeneration and destroy in OA model rabbits in time and dose dependently. Its mechanism may be associated with expression up-regulation of Bcl-2 and expression down-regulation of Bax.
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OBJECTIVE:To study the effects of Cornus officinalis polysaccharide on learning and memory ability of vascular dementia (VD) model rats and its mechanism. METHODS:30 rats were randomly divided into sham operation (normal saline) group,model(normal saline)group and C. officinalis polysaccharide(0.28 g/kg)group. Except sham operation group,VD model was induced in other groups and given relevant medicine intragastrically once a day. After 4 weeks,the frequency of leaping over original platform position,water maze mean escape latencies,platform retention time and average swimming distance were detect-ed,and the mRNA and protein expression of brain-derived neurotrophic factor(BDNF)and apoptosis related gene Bcl-2 in hippo-campal tissue of rats were determined. RESULTS:Compared with sham operation group,learning and memory ability of rats in model group weakened,manifesting as the frequency of leaping over original platform position decreased,water maze mean escape latencies prolonged,platform retention time shortened,average swimming distance extended(P<0.05);mRNA and protein expres-sion of BDNF and Bcl-2 in hippocampus decreased significantly (P<0.05). Compared with model group,learning and memory ability of rats in C. officinalis polysaccharide group increased,manifesting as the frequency of leaping over original platform posi-tion increased,water maze mean escape latencies shortened,platform retention time prolonged,average swimming distance short-ened(P<0.05);mRNA and protein expression of BDNF and Bcl-2 in hippocampus was enhanced significantly(P<0.05). CON-CLUSIONS:C. officinalis polysaccharide can improve the learning and memory ability of VD rats,and its mechanism may be asso-ciated with the expression up-regulation of BDNF and Bcl-2 in hippocampus.
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OBJECTIVE:To study the effects of podophyllotoxin derivative QW-83 on human cervical cancer HeLa cell apopto-sis and its mechanism. METHODS:After treated with 0(negative control),0.01,0.1,1 and 10 μmol/L QW-83 and positive drug etoposide(VP-16)for 48 h,proliferation inhibition rate and IC50 of HeLa cell were determined by MTT assay. The morphological changes of HeLa cell were observed by Hochest 33342 staining after treated with QW-83 [0(negative control),2.5,5,10μmol/L] for 48 h;flow cytometry was used to detect apoptosis rate;semi quantitative RT-PCR was adopted to detect the expression of apop-tosis related gene P53,Bax,Casepase-3,Casepase-8,Casepase-9 and Bcl-2 mRNA. RESULTS:Compared with negative control, 1,10 μmol/L VP-16 and QW-83 had obvious proliferation inhibition effect on HeLa cells (P<0.05 or P<0.01),and IC50 were (5.11±0.43)μmol/L and(4.96±0.54)μmol/L. Hochest 33342 staining results showed QW-83 could obviously induce cells apopto-sis and nuclear pyknosis. Flow cytometry showed QW-83 could increase apoptosis rate in concentration-dependent manner,being 16.89%-62.56%. RT-PCR showed mRNA expression of P53,Bax,Caspase-3,Casepase-8 and Casepase-9,Bcl-2/Bax increased, while mRNA expression of Bcl-2 decreased after treated with QW-83(P<0.05). CONCLUSIONS:Podophyllotoxin derivative QW-83 can induce HeLa cell apoptosis,and its mechanism may be associated with regulate mRNA expression of apoptosis related gene.
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Background Pterygium is a relatively common eye disease,but its aetiology and pathogenesis remain uncertain.At present,the study on pterygia focuses on understanding its underlying mechanism.Objective This study was to detect the expression of 8-hydroxy-2'-deoxyguaine (8-OHdG),a marker of oxidative damage of DNA,and bcl-2,a gene related with apoptosis,on the pterygium tissue.Methods Thirty pterygium tissue specimens were obtained during the surgery with the primary pterygium 24 cases and recurrent pterygium 6 cases.In addition,20 normal conjunctival specimens from retinal detachment surgery and strabismus surgery were collected.The expressions of 8-OHdG and bcl-2 in pterygium tissue were detected using immunochemistry and compared with the normal conjunctival tissue.The difference in the expressions of 8-OHdG and bcl-2 among different specimens was compared by x2test,and the relationship between 8-OHdG expression and bcl-2 expression was evaluated by Kappa test.Results The positive expressing rate of 8-OHdG in the pterygium tissue was 62.5％ and 83.3％ in the primary and recrudescence pterygium tissue,respectively,but the expression of 8-OHdG was absent in the normal conjunctiva tissue.No significant difference was found in the positive expressing rate of 8-OHdG between primary and recrudescence pterygium tissue(x2 =0.938,P＞0.05).The bcl-2 expressing rate was 90.0％ and 87.5％ in the primary and recrudescence pterygium tissue,respectively.However,that in the normal conjunctival tissue was absent.No significant difference was seen in the bcl-2 expression rate between primary and recrudescence pterygium tissue (x2=0.833,P ＞ 0.05).Of the 27 pterygium tissue with bcl-2 positive expression,8-OHdG showed the positive expression in 20 specimens,and 3 specimens with the bcl-2 negative response were absent reactive to 8-OHdG,showing insignificant difference between them (P＞0.05).The relationship between 8-OHdG expression and bcl-2 expression was concord in a certein extent (Kappa =0.464).Conclusions The upregulation of 8-OHdG in the pterygium tissue indicates that oxidative damage of DNA plays a role in the development of pterygium.Oxidative damage of DNA caused by ultraviolet may be an upriver factor,which induces raising up of expression of bcl-2 and inhibits the apoptosis of normal cells and further proliferation of the conjunctiva tissue,resulting in the genesis and development of pterysium.
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AIM:To observe the effects of recombi- nant human endostatin(rh-end)on the expression of p53,fas and bcl-2 genes in synovial tissue in rats with adjuvant arthritis(AA).METHODS:The effects of rh- end on the expression of p53,fas and bcl-2 mRNA in synovial tissue in AA rats were examined quantitatively by SYBR GreenⅠreal-time fluorescent quantitative PCR (FQ-PCR).RESULTS:The expression offas and bcl-2 mRNA in synovial tissue in AA rats treated with rh-end was significantly increased,as compared with model con- trois(P
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AIM: To clarify the role of new apoptosis-related gene, TF-1 cell apoptosis-related gene 19(TFAR19), in the pathogenesis of systemic lupus erythematosis (SLE) and the relationship between TFAR19 and SLE. METHODS: DNA Ladder detection, Western blotting, immunological fluorescence method, ELISA and so on were used to test if ultraviolet B(UVB) could induce HaCaT cell apoptosis and TFAR19 expression. RESULTS: HaCaT cell apoptosis could be detected after 24 hours of 30 mj/cm 2 UVB irradiation. Also, we found that in active SLE patients, the TFAR19 antibody was increased, but not significant compared to the normal control. CONCLUSION: TFAR 19 is involved in the process of UVB induced ketatinocyte line HaCaT apoptosis and SLE pathogenesis.