Sun Shou-Li, Guo Li, Ren Ya-Chao, Wang Bing, Li Rong-Hui, Qi Yu-Shan, Yu Hui, Chang Nai-Dan, Li Ming-Hui, Peng Hai-Sheng
Daqing Campus, Harbin Medical University, Qaqing, China.
Mol Biol Rep. 2014 Sep;41(9):6117-24. doi: 10.1007/s11033-014-3490-1. Epub 2014 Jun 28.
To investigate the mechanism of apoptosis in myocardial cells of aging rats induced by D-galactose and to study the effect of the Polysaccharide isolated from the seeds of Cuscuta chinensis Lam (PCCL) on apoptosis of cardiomyocytes and its corresponding machinasim in aging rat model. Fifty male SD rats were randomly divided into 5 groups. Normal control group (NC). D-galactose (100 mg · kg(-1)d(-1) for 56 day) indued aging group (MC), D-galactose plus 100 mg kg(-1) d(-1) PCCL group (ML), D-galactose plus 200 mg kg(-1) d(-1) PCCL group (MM), and D-galactose plus 400 mg kg(-1) d(-1) PCCL group (MH). Same volume of solution (water, or PCCL aqueous solution) was given by gavage for 56 days. Then the hearts were collected and apoptosis parameters were evaluated. Caspase-3 and Cyt c were determined by fluorescence spectrometer, the apoptosis rate was assessed by AnnexinV-FITC method by Flow-Cytometry, [Ca(2+)]i and [Ca(2+)]i overloaded by KCL were observed by laser scanning confocal microscopy (LSCM); Bcl-2 and Bax were examined by immunohistochemistry. The content of Cyt C, [Ca(2+)]i of cardiomyocytes, the activity of Caspase-3, Bax expression level in D-galactose induced aging group were higher than NC (p < 0.05). The ratio of Bcl-2/Bax was decreased in D-galactose induced aging group compared to NC. On the other hand, the content of Cyt C, [Ca(2+)]i of cardiomyocytes, the activity of Caspase-3 and apoptosis rate, as well as Bax expression level in all three PCCL groups were decreased compared to galactose induced group (p < 0.05). Bcl-2/Bax ratio was increased in all PCCL groups compared to galactose induced aging group. PCCL could decrease the apoptosis of cardiomyocytes by the mitochondria apoptosis pathway.
探讨D-半乳糖诱导衰老大鼠心肌细胞凋亡的机制,研究菟丝子种子多糖(PCCL)对衰老大鼠模型心肌细胞凋亡的影响及其相应机制。将50只雄性SD大鼠随机分为5组。正常对照组(NC)、D-半乳糖(100 mg·kg⁻¹d⁻¹,共56天)诱导衰老组(MC)、D-半乳糖加100 mg kg⁻¹ d⁻¹ PCCL组(ML)、D-半乳糖加200 mg kg⁻¹ d⁻¹ PCCL组(MM)和D-半乳糖加400 mg kg⁻¹ d⁻¹ PCCL组(MH)。通过灌胃给予相同体积的溶液(水或PCCL水溶液),持续56天。然后收集心脏并评估凋亡参数。用荧光光谱仪测定Caspase-3和细胞色素C(Cyt c),采用流式细胞术通过AnnexinV-FITC法评估凋亡率,用激光扫描共聚焦显微镜(LSCM)观察[Ca²⁺]i和KCl诱导的[Ca²⁺]i超载;用免疫组织化学法检测Bcl-2和Bax。D-半乳糖诱导衰老组心肌细胞的Cyt C含量、[Ca²⁺]i、Caspase-³活性、Bax表达水平均高于NC组(p<0.05)。与NC组相比,D-半乳糖诱导衰老组Bcl-2/Bax比值降低。另一方面,与半乳糖诱导组相比,所有三个PCCL组心肌细胞的Cyt C含量、[Ca²⁺]i、Caspase-³活性和凋亡率以及Bax表达水平均降低(p<0.05)。与半乳糖诱导衰老组相比,所有PCCL组的Bcl-2/Bax比值均升高。PCCL可通过线粒体凋亡途径减少心肌细胞凋亡。