RESUMO
OBJECTIVE: To investigate cell apoptosis, mitochondrial membrane potential, cytochrome C and mechanisms of mitochondria in lymphocyte apoptosis of sepsis. METHODS: In the research, female C57BL/6 mice whose body weight ranged from 17 to 25 grams were utilized and assigned randomly to two groups: sham operated group (Control), cecal ligation and puncture group (CLP). The present study was undertaken by using the mice splenic lymphocyte to investigate cell apoptosis, mitochondrial membrane potential, cytochrome C. The apoptosis alteration was evaluated by Annexin V-FITC/PI double staining with flow cytometry. The alteration of mitochondrial membrane potential was investigated by Rhodamine-123 staining of cells. Cytochrome C of mitochondria and cytosol was investigated by Western blot methods. Statistical analysis was performed using SPSS 11.5 for Windows software. Experiment data was indicated with mean ± standard. RESULTS: The splenic lymphocyte apoptosis was significantly accelerated in the CLP group when compared with that in control group (17.3% ± 2.2% vs. 3.5% ± 0.5%, P < 0.05). The Rhodamine-123 fluorescent intensity in splenic lymphocyte apoptosis was reduced in CLP group (76.2% ± 1.6%). Comparison between sham group (99.6% ± 0.4%) and CLP group had statistical significance (P < 0.05). Apoptosis could induce mitochondrial cytochrome C release into cytoplasm. In the CLP group, elevation of cytochrome C in cytosol was concurrently in accordance with decline in mitochondrial cytochrome C content. CONCLUSION: These data suggest that mitochondria and mitochondria signal pathway play an important role in lymphocyte apoptosis of sepsis.
Assuntos
Citocromos c/metabolismo , Linfócitos/patologia , Mitocôndrias/fisiologia , Sepse/patologia , Animais , Apoptose , Células Cultivadas , Modelos Animais de Doenças , Feminino , Linfócitos/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Mitocôndrias/metabolismo , Sepse/metabolismoRESUMO
OBJECTIVE: To investigate the role of endoplasmic reticulum (ER) stress-mediated apoptosis signal pathway in extensive apoptosis of splenic lymphocytes in sepsis. METHODS: Twenty-four C57BL/6 mice were randomized into the cecal ligation and puncture (CLP) group and sham operation group, with 12 mice in each group. Twenty-four hours after CLP, the animals were sacrificed, and spleens were harvested for evaluation. Apoptosis was detected by the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) analysis. The expressions of glucose-regulated protein 78 (GRP78) and C/EBP homologous protein (CHOP) were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting. RESULTS: High degree of lymphocyte apoptosis was observed in the CLP group. Meanwhile, marked induction of GRP78 expression was observed in the splenocytes in septic mice, indicating the motivation of the unfolded protein responses (UPR). Furthermore, both mRNA and protein levels of CHOP were highly upregulated in the CLP group, suggesting that ER stress response was switched to a pro-apoptotic response. CONCLUSION: These results demonstrates activation of the ER stress response in lymphocytes and that ER stress may contribute to abnormal lymphocyte apoptosis during sepsis.
Assuntos
Retículo Endoplasmático/metabolismo , Linfócitos/patologia , Sepse/metabolismo , Transdução de Sinais , Baço/patologia , Animais , Apoptose , Modelos Animais de Doenças , Chaperona BiP do Retículo Endoplasmático , Proteínas de Choque Térmico HSP70/genética , Proteínas de Choque Térmico HSP70/metabolismo , Linfócitos/metabolismo , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , RNA Mensageiro/genética , Distribuição Aleatória , Sepse/patologia , Baço/metabolismo , Fator de Transcrição CHOP/genética , Fator de Transcrição CHOP/metabolismoRESUMO
In recent years, China has constantly strengthened environmental regulation (ER) to force the manufacturing industry to upgrade. This study theoretically analyzes interaction mechanism of ER on the upgrading of manufacturing industry through foreign direct investment (FDI) and technological innovation (TI) and carries out empirical verification by using provincial panel data from 2000 to 2016 in China. The results demonstrate that the current ER intensity in China is unable to directly promote the upgrading of manufacturing industry, while through the interaction effects of FDI and TI do boost the upgrading of the industry. The above mechanisms are also robust even if we take the regional heterogeneity into consideration. Basic education and urbanization are favorable for the upgrading of the manufacturing industry. However, the increase in dependence on foreign trade is not conducive to upgrading manufacturing industry. Chinese government should further strengthen ER and give full play of the interaction mechanism of ER to guide the flow of foreign investment and force enterprises to carry out TI. In the meanwhile, Chinese government also needs to ensure balanced regional development, thus better promoting the upgrading of manufacturing industry.