目的研究SIRT3在不同热量饮食下大鼠肾脏的表达变化规律。方法取12月龄自由饮食、热量限制、高热量饮食、健康雄性Fisher344大鼠肾组织进行衰老相关β-半乳糖苷酶(SA-β-gal)活性染色,Western印迹法(Western blot assay)和免疫组织化学...目的研究SIRT3在不同热量饮食下大鼠肾脏的表达变化规律。方法取12月龄自由饮食、热量限制、高热量饮食、健康雄性Fisher344大鼠肾组织进行衰老相关β-半乳糖苷酶(SA-β-gal)活性染色,Western印迹法(Western blot assay)和免疫组织化学法检测肾脏组织中SIRT3的表达变化与定位,并用Western blot检测p16INK4A在肾脏组织中的表达变化。结果大鼠肾脏组织SA-β-gal活性随饮食中热量的增长逐渐增强(P<0.05);p16INK4A表达随饮食中热量增加逐渐增强(P<0.05)。Western blot亦显示SIRT3蛋白表达随饮食中热量的增加逐渐减弱(P<0.05)。免疫组化结果显示,SIRT3蛋白主要表达于大鼠肾小管中,其在肾小球也有微量表达,且跟Western blot结果一致,随饮食中热量增长表达增加(P<0.05)。结论热量限制能延缓大鼠肾脏衰老,SIRT3可能与高热量引起的肾脏衰老有密切关系。展开更多
This study was designed to investigate the effect of 3 D TECA hydrogel on the inflammatory-induced senescence marker, and to assess the influence of the gel on the periodontal lig-ament fibroblasts(PDLFs) migration in...This study was designed to investigate the effect of 3 D TECA hydrogel on the inflammatory-induced senescence marker, and to assess the influence of the gel on the periodontal lig-ament fibroblasts(PDLFs) migration in wound healing in vitro. PDLFs were cultured with20 ng/ml TNF-α to induce inflammation in the presence and absence of 50 μM 3 D TECA gelfor 14 d. The gel effect on the senescence maker secretory associated-β-galactosidase(SA-β-gal) activity was measured by a histochemical staining. Chromatin condensation and DNAsynthesis of the cells were assessed by 4,6-diamidino-2-phenylindole and 5-ethynyl-2-deoxyuridine fluorescent staining respectively. For evaluating fibroblasts migration, scratchwound healing assay and Pro-Plus Imaging software were used. The activity of senescencemarker, SA-β-gal, was positive in the samples with TNF-α-induced inflammation. SA-β-galpercentage is suppressed( > 65%, P < 0.05) in the treated cells with TECA gel as compared tothe non-treated cells. Chromatin foci were obvious in the non-treated samples. DNA syn-thesis was markedly recognized by the fluorescent staining in the treated compared to non-treated cultures. Scratch wound test indicated that the cells migration rate was significantlyhigher(14.9 μm^2/h, P < 0.05) in the treated versus(11 μm^2/h) for control PDLFs. The new for-mula of 3 D TECA suppresses the inflammatory-mediated cellular senescence and enhancedfibroblasts proliferation and migration. Therefore, 3 D TECA may be used as an adjunct to ac-celerate repair and healing of periodontal tissues.展开更多
基金support from the Ministry of Science, Technology and Innovation for provid-ing the grant number (MOSTI SF01.01.09)the Ministryof Higher Education for providing the Research AcculturationGrant Scheme (RAGS 119/2013)UiTM DANA (RIF 751/2012)
文摘This study was designed to investigate the effect of 3 D TECA hydrogel on the inflammatory-induced senescence marker, and to assess the influence of the gel on the periodontal lig-ament fibroblasts(PDLFs) migration in wound healing in vitro. PDLFs were cultured with20 ng/ml TNF-α to induce inflammation in the presence and absence of 50 μM 3 D TECA gelfor 14 d. The gel effect on the senescence maker secretory associated-β-galactosidase(SA-β-gal) activity was measured by a histochemical staining. Chromatin condensation and DNAsynthesis of the cells were assessed by 4,6-diamidino-2-phenylindole and 5-ethynyl-2-deoxyuridine fluorescent staining respectively. For evaluating fibroblasts migration, scratchwound healing assay and Pro-Plus Imaging software were used. The activity of senescencemarker, SA-β-gal, was positive in the samples with TNF-α-induced inflammation. SA-β-galpercentage is suppressed( > 65%, P < 0.05) in the treated cells with TECA gel as compared tothe non-treated cells. Chromatin foci were obvious in the non-treated samples. DNA syn-thesis was markedly recognized by the fluorescent staining in the treated compared to non-treated cultures. Scratch wound test indicated that the cells migration rate was significantlyhigher(14.9 μm^2/h, P < 0.05) in the treated versus(11 μm^2/h) for control PDLFs. The new for-mula of 3 D TECA suppresses the inflammatory-mediated cellular senescence and enhancedfibroblasts proliferation and migration. Therefore, 3 D TECA may be used as an adjunct to ac-celerate repair and healing of periodontal tissues.
