清感冬饮预防性干预对系统性红斑狼疮合并感冒小鼠T淋巴细胞的影响OA

Objective To investigate the effect of prophylactic application of Qinggan Dongyin Decoction(QGDY)on T lymphocytes in systemic lupus erythematosus(SLE)mice following a simulated common cold,using an SLE-complicated-with-cold mouse model.Methods Eighteen female MRL/lpr mice were randomly divided into three experimental groups(n=6 per group):the model group,the simulated common cold group,and the QGDY prevention group.Additionally,six age-matched female C57BL/6 mice served as the control group.The QGDY prevention group received daily intragastric administration of QGDY(17.94 g/kg),while the other three groups received an equal volume of saline.This administration continued for 2 consecutive weeks.Subsequently,the simulated common cold group and the QGDY prevention group were subjected to a mouse cold model,while the control and model groups were housed at room temperature.One week later,samples were collected.Lung and kidney tissues were processed for hematoxylin-eosin(HE)staining,and pathological changes were observed under light microscopy.Protein expression levels of interferon regulatory factor 7(IRF7)and phosphorylated IRF7(p-IRF7)in lung tissue were detected by western blot.IRF7 mRNA expression in lung tissue was measured by Real-time fluorescent quantitative polymerase chain reaction(RT-qPCR).Flow cytometry was used to assess the percentages of CD4+T cells,CD8+T cells,and the proportions of Th1,Th2,and Treg cell subsets in splenic tissue.Serum levels of tumor necrosis factor-alpha(TNF-α),interleukin-6(IL-6),and perforin(PFP)were measured by enzyme-linked immunosorbent assay(ELISA)using blood samples collected via orbital bleeding.Results(1)Body temperature of mice in each group was measured at different time points.Results showed a significant interaction between group and time[F(9,60)=11.505,P<0.01],indicating different trends of temperature change among groups.At the 3rd week,the body temperature in the simulated common cold group was significantly higher than that in the model group(P<0.01).The body temperature in the simulated common cold group at the 3rd week was significantly higher than that at week 0,week 1,and week 2 within the same group(all P<0.01).In the QGDY prevention group,body temperature at the 3rd week was significantly higher than that at week 0,week 1,and week 2(all P<0.05).No statistically significant differences in body temperature were observed at any time point in the control group or the model group(P>0.05).(2)HE staining:In the control group,alveolar distribution was uniform and cells were arranged neatly.Compared with the control group,the model group showed disordered cell arrangement,enhanced pulmonary parenchymatous changes,and aggravated inflammation.Compared with the model group,the simulated common cold group exhibited more disordered cell arrangement,further enhanced pulmonary parenchymatous changes,aggravated inflammation,and increased hemorrhagic spots.Compared with the simulated common cold group,the QGDY prevention group showed relatively neat cell arrangement,reduced pulmonary parenchymatous changes,and alleviated inflammation.In the control group,renal glomeruli were intact in morphology and structure with minimal inflammation.Compared with the control group,the model group displayed glomerular shrinkage,vacuolation,hyperplasia,and obvious inflammatory cell infiltration.Compared with the model group,the simulated common cold group showed aggravated glomerular shrinkage,vacuolation,and inflammation.Compared with the simulated common cold group,the QGDY prevention group exhibited relatively well-organized glomerular structure and reduced inflammation.(3)WB:Compared with the control group,IRF7 expression was increased in the model group(P<0.05),while no statistically significant difference were observed in p-IRF7 and p-IRF7/IRF7 ratio(P>0.05).Compared with the model group,p-IRF7 expression was increased in the simulated common cold group(P<0.05),while no statistically significant difference were observed in IRF7 and p-IRF7/IRF7 ratio(P>0.05).Compared with the simulated common cold group,no statistically significant difference were observed in IRF7,p-IRF7,or p-IRF7/IRF7 ratio in the QGDY prevention group(P>0.05).(4)RT-qPCR:Compared with the control group,no statistically significant difference was observed in IRF7 mRNA expression in the model group(P>0.05).Compared with the model group,IRF7 mRNA expression was increased in the simulated common cold group(P<0.01).Compared with the simulated common cold group,no statistically significant difference was observed in IRF7 mRNA expression in the QGDY prevention group(P>0.05).(5)Flow cytometry:Compared with the control group,the model group showed decreased expression of CD4+T and CD8+T cells(both P<0.01),decreased Treg cells(P<0.01),increased Th1 and Th2 cells(both P<0.01),and no statistically significant difference in Th1/Th2 ratio(P>0.05).Compared with the model group,the simulated common cold group showed no statistically significant difference in CD4+T,CD8+T,Treg,or Th2 cells(P>0.05),but decreased Th1 cells(P<0.05)and a decreased Th1/Th2 ratio(P<0.01).Compared with the simulated common cold group,the QGDY prevention group showed increased expression of CD4+T and CD8+T cells(both P<0.01),no statistically significant difference in Treg and Th1 cells(P>0.05),decreased Th2 cells(P<0.01),and an increased Th1/Th2 ratio(P<0.01).(6)ELISA:Compared with the control group,the model group showed no statistically significant difference in IL-6(P>0.05),increased expresion of TNF-α(P<0.05)and decreased expression of PFP(P<0.01).Compared with the model group,the simulated common cold group showed no statistically significant difference in IL-6,TNF-α,or PFP(P>0.05).Compared with the simulated common cold group,the QGDY prevention group showed no statistically significant difference in IL-6,TNF-α,or PFP(P>0.05).Conclusion The prophylactic administration of QGDY may alleviate inflammatory cell infiltration in the lung and kidney tissues of MRL/lpr mice after cold induction,potentially through the regulation of T-cell subsets.