ABSTRACT
Aim: To investigate the effect of salvianolic acid B (Sal B) on liver fibrotic cells in vivo and in vitro from die perspective of apoptosis, as well as the effect on cleaved caspase-9. Methods: Diethylnitrosamine (DEN) was used to induce liver fibrosis in mice for 12 weeks. The padiological changes were detected by HE staining, and the fibrotic lesion area was determined. The cell apoptosis in the fibrotic area was observed by Hoechst 33258 fluorescence staining. The expression of cleaved caspase-9 in fibrotic tissues was detected by Western blot. The apoptotic rate of each group was detected by double standard method AnnexinV-FITC/PI, and the expression of apoptotic protein cleaved caspase-9 in HSC-T6 was detected by Western blot. Results: HE staining suggested that 12 weeks were the period of liver fibrosis in mice. No pseudoplobular structure was formed in group with low and high dose of Sal B, and the degree of fibrosis was lower than that in model group. In the fibrotic lesion area, the fluorescence staining of Hoechst 33258 showed that apoptotic cells significantly increased in group with low Sal B and high dose compared with model group. The results of the AnnexinV-FITC/PI method showed that TGF-β1 inhibited the apoptosis of HSC-T6, and Sal B promoted the apoptosis of HSC-T6 after TGF-β1 intervention and showed a concentration dependence (P <0. 01). Western blot results showed that in fibrotic liver tissues, Sal B increased the expression of cleaved caspase-9 in HSC-T6 cells compared with model group. Compared with TGF-β1 group, Sal B increased cleaved caspase-9 protein expression (P < 0. 01). Conclusions: Sal B can significantly promote apoptosis of liver fibrotic cells in vitro and in vivo, and its pro-apoptosis mechanism may be related to the up-regulation of cleaved caspase-9.
ABSTRACT
Objective To explore the effect of acupuncture-rehabilitation therapy on apoptosis and the expression of X-linked inhibitor of apoptosis protein (XIAP) and cleaved-caspase-9 in ischemic penumbra of rats with cerebral ischemia. Methods A total of 180 male Sprague-Dawley rats were randomly divided into sham operation group,model group,acupuncture group,rehabilitation group and acupunc-ture-rehabilitation therapy group.Each group was divided into three days,seven days and 14 days subgroups(n=12).The model was estab-lished with the modified Longa suture method. The model group and the sham operation group received no treatment. The acupuncture group received cluster needling of scalp acupuncture,the rehabilitation group received treadmill training,and the acupuncture-rehabilitation therapy group received both acupuncture and treadmill training.On the third,seventh and 14th days after modeling,the neurological func-tion was assessed with modified Neurological Severity Score(mNSS).The apoptosis was observed by TUNEL staining and the expression of XIAP and cleaved-caspase-9 protein was determined by Western blotting in cerebral ischemic penumbra,respectively.Results Compared with the model group,the mNSS scores decreased,the rate of apoptosis decreased,the expression of XIAP protein increased,and the expres-sion of cleaved-caspase-9 decreased in each treatment group at each time point(P<0.05).Compared with the other two treatment groups,the rate of apoptosis further decreased at each time points(P<0.05),the expression of XIAP protein further increased at each time points(P<0.05),the mNSS score further decreased(P<0.05)and the expression of cleaved-caspase-9 protein further decreased(P<0.05)seven days and 14 days after modeling in the acupuncture-rehabilitation therapy group.Conclusion Acupuncture and rehabilitation therapy could re-duce the neurological deficit in rats with cerebral ischemia,which was superior to the simple acupuncture treatment and rehabilitation train-ing.The mechanism may be related to th reduction of apoptosis by promoting XIAP protein expression and inhibiting caspase-9 protein acti-vation.