الملخص
Objective: T lymphocyte exhaustion is an important component of immune dysfunction. Therefore, exploring peripheral blood-exhausted T lymphocyte features in patients with hepatitis B virus-related acute-on-chronic liver failure may provide potential therapeutic target molecules for ACLF immune dysfunction. Methods: Six cases with HBV-ACLF and three healthy controls were selected for T-cell heterogeneity detection using the single-cell RNA sequencing method. In addition, exhausted T lymphocyte subpopulations were screened to analyze their gene expression features, and their developmental trajectories quasi-timing. An independent sample t-test was used to compare the samples between the two groups. Results: Peripheral blood T lymphocytes in HBV-ACLF patients had different differentiation trajectories with different features distinct into eight subpopulations. Among them, the CD4(+)TIGIT(+) subsets (P = 0.007) and CD8(+)LAG3(+) (P = 0.010) subsets with highly exhausted genes were significantly higher than those in healthy controls. Quasi-time analysis showed that CD4(+)TIGIT(+) and CD8(+)LAG3(+) subsets appeared in the late stage of T lymphocyte differentiation, suggesting the transition of T lymphocyte from naïve-effector-exhausted during ACLF pathogenesis. Conclusion: There is heterogeneity in peripheral blood T lymphocyte differentiation in patients with HBV-ACLF, and the number of exhausted T cells featured by CD4(+)TIGIT(+)T cell and CD8(+)LAG3(+) T cell subsets increases significantly, suggesting that T lymphocyte immune exhaustion is involved in the immune dysfunction of HBV-ACLF, thereby identifying potential effective target molecules for improving ACLF patients' immune function.
الموضوعات
Humans , Hepatitis B virus , Acute-On-Chronic Liver Failure/pathology , Hepatitis B, Chronic , T-Lymphocyte Subsets/pathology , Receptors, Immunologicالملخص
Objective To investigate the effects of over-expression ofhypoxia inducible factor 1α (HIF-1α) on apoptosis of PC12 cells under hypoxic injury condition mimicked by CoCl2. Methods To obtain the neural cell line over-expressing HIF-1α, the recombinant eukaryotic expression vector of pEGFPC1-HIF-1α△ODD was transfected into the differentiated PC12 cells, and pEGFPC1 was transfected into the PC12 cells as control group. PC12 cells under simulative hypoxic injury condition was constructed by CoCl2 treatment. The expression of HIF-1α was analyzed under normal and hypoxia conditions, respectively, using the method of Western blotting. Then Hoechst33342 staining, flow cytometer and caspase-3 activity detection were employed to measure and compare the cell apoptosis rates between the over-expressing HIF-1α cell line and the controls. Results We successfully constructed the model with simulative hypoxic injury to the PC12 cell line via the treatment of 50 μ mol/L or 100 μ mol/L CoCl2 for 12-24 h. The results from Western blotting illustrated that cells transfected by pEGFPC1-HIF-1α△ODD highly expressed HIF-1α under both normal and hypoxic conditions as compared with the control cells. Furthermore, Hoechst33342 staining indicated that the apoptosis rate in those transfected by pEGFPC1-HIF-1α△ODD was significantly decreased as compared with that in the control cells; the apoptosis rate in those transfected by pEGFPC1-HIF-1α△ODD (5.41±3.29)% and the control cells (8.35±2.59)% was obviously different 24 h after treatment with 50 μ mol/L CoCl2 (P＜0.05).The caspase-3 activities were significantly different between those transfected by pEGFPC1-HIF-1α△ODD (2.152±0.384) and control cells (0.921 ±0.287) 12 h after treatment with 50μmol/L CoCl2 (P＜0.05). Conclusion In the model of neural cells undergoing simulative hypoxic injury by the treatment of CoCl2, over-expression of HIF-1α can inhibit the cell apoptosis.
الملخص
Objective To investigate the activity change pattern and corresponding significance of gly- colytic enzymes including alsolase A(ALDA)and lactate dehydrogenase M(LDH-M)regulated by hy- poxia-inducible factor 1?(HIF-1?)in spinal cord injury(SCI)of rats.Methods SD rats were ran- domly divided into control group and groups at 12 hours,1,2 and 3 days,1 and 2 weeks after compres- sive SCI,in which the activity changes of ALDA and LDH-M in the injured spinal cord were observed at different time points by means of enzyme histochemistry.Results Opitical density(A)value of AL- DA continued significant increase from two days to one week after SCI(P＜0.05)and decreased gradual- ly at 2 weeks after SCI.A value of LDH-M began significant increase at day 1 after SCI and recovered to normal level at 2 weeks after SCI(P＜0.05).Conclusion Activities of ALDA and LDH-M regulated by HIF-1?in spinal cord injury is significantly increased.