Effects of scutellarin on neuroinflammation in rats with traumatic brain injury by regulating cGAS/STING signaling pathway / 中国药房

OBJECTIVE To investigate the improvement effects and mechanism of scutellarin （Scu） on neuroinflammation in rats with traumatic brain injury （TBI）. METHODS The modified Feeney method was applied to construct TBI rat model. The rats were randomly grouped into TBI group，Scu low-dose group （40 mg/kg），Scu high-dose group （80 mg/kg），cyclic guanylate- adenylate synthase （cGAS） inhibitor group （cGAS inhibitor RU.521，450 μg/kg），with 24 rats in each group. Other 24 rats were included in the sham operation group. The modified neurological deficit score （mNSS） method was applied to assess the neurological function of rats； the brain water content of rats was measured by dry/wet specific gravity method； hematoxylin-eosin and TdT-mediated dUTP nick-end labeling staining were applied to observe the pathological changes and apoptosis of brain tissue in rats； the levels of interferon-β （IFN-β），CXC chemokine ligand-10 （CXCL10），tumor necrosis factor-α （TNF-α） and interleukin-6 （IL-6） in rat brain tissue were detected by enzyme-linked immunosorbent assay； Western blot method was applied to detect the expression of cGAS/interferon gene stimulating protein （STING） signal pathway-related proteins in brain tissue of rats. RESULTS Compared with the sham operation group，the mNSS，brain water content，apoptosis rate，the contents of IFN-β，CXCL10，TNF-α and IL-6，and the relative expressions of cGAS and STING proteins in TBI group increased significantly （P＜0.05）； there were edema，bleeding and pathological damage to neurons in the brain tissue. Compared with TBI group，the above indicators and pathological changes of rats in administration groups were improved significantly （P＜0.05），and the effect of Scu was in a dose- dependent manner （P＜0.05）； however，there was no statistically obvious difference in the above indicators between the Scu high- dose group and the cGAS inhibitor group （P＞0.05）. CONCLUSIONS Scu may alleviate neuroinflammation，reduce brain tissue damage and apoptosis，and promote the recovery of neural function in TBI rats by inhibiting the activation of cGAS/STING signaling pathway.

The radiosensitization effect of STING agonist on cutaneous melanoma cells / 中华放射医学与防护杂志

Objective:To investigate the radiosensitizing effect and underlying mechanism of STING agonist (c-di-AMP) on cutaneous melanoma cells.Methods:Human cutaneous melanoma cells (A375) were divided into four groups: the control group, 10 μmol/L c-di-AMP group, X-ray irradiation group and X-ray irradiation combined with c-di-AMP group. The radiosensitizing effect of c-di-AMP on A375 cells was detected by CCK-8-based viability assay, lactate dehydrogenase (LDH) release assay, flow cytometry-based apoptosis assay, and colony formation assay. Western blot analysis was used to determine the expressions of cell death-related proteins.Results:In combination with 10 Gy X-ray irradiation, 10 μmol/L c-di-AMP showed significant radiosensitization effect in A375 cells, which was evidenced by decreased cell activity ( t=5.11, P<0.05), increased cytotoxicity ( t=10.15, P<0.05) and cell apoptosis ( t=4.41, P<0.05) and reduced clone viability( t=6.30, 3.55, 5.45, 3.55, P<0.05). The calculated radiosensitization ratio of c-di-AMP to A375 cells was 1.88. Moreover, 10 μmol/L c-di-AMP further increased the expressions of cell death-related proteins induced by radiation in A375 cells. Conclusions:The STING agonist c-di-AMP can be used as a radiosensitizer for cutaneous melanoma, which may provide a novel strategy for radiotherapy.

Relationship between STING gene expression and HBV DNA load in peripheral blood mononuclear cells of patients with chronic hepatitis B / 中国感染控制杂志

Objective To detect mRNA expression of stimulator of interferon genes(STING)and type Ⅰ interferons(IFN-α and IFN-β)in peripheral blood mononuclear cells(PBMCs)of patients with chronic hepatitis B(CHB), and evaluate its correlation with hepatitis B virus load.Methods 88 untreated CHB patients(CHB group)and 74 healthy persons(control group)who performed physical examination were chosen from Renmin Hospital of Wuhan University during the same period between February 2016 and February 2017.Expressions of mRNA of STING, IFN-α, and IFN-βwere detected by quantitative real-time polymerase chain reaction(PCR), their relative expression values were obtained by 2-ΔΔCT method, results were statistically analyzed.Results The expression of STING, IFN-α, and IFN-βmRNA in peripheral blood of CHB patients were 2.95, 3.14, and2.01folds of healthy controls respectively, differences were statistically significant(t=-4.72, -3.41, -2.31, respectively, all P＜0.05).STING relative expression in patients with HBV DNA load≤104 IU/mL was 2.98, 3.76, and 3.97 folds of patients with HBV DNA load 104-105 IU/mL, 105-106 IU/mL, and＞106 IU/mL, respectively(P＜0.05).mRNA expressions of STING in CHB patients were positively correlated with that of IFN-αand IFN-βmRNA (r=0.475, 0.503, respectively, both P＜0.05).Conclusion The expression of STING increased in patients with CHB, high expression of STING impacted the replication of HBV.

STUDY ON INTERFERON-? GENE-MODIFIED HUMAN TUMOR CELLS / 中国免疫学杂志

The whole length cDNA of human interferon-?(IFN-?) gene including signal peptide was constructed to retroviral vector pLXSN and then packaged with PA317. A Human Hepetoma Cell Line(HHCL) and a human gastric cancer cell line (MKN-45) were infected by using supernatant containing retroviral vector with IFN-?. the gene-modified tumor cell. were isolated by G418 selection. Some changes were found in cell cycle, expression of HLA class Ⅰ and classⅡ, and tumorigenecity. Elevated immunogenecity and abrogated rumerigenecity suggest a means for generating gene-modified tumor cell vaccine. for the treatment of cancer.