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ObjectiveTo explore the optimal formula of Maxing Shigantang in regulating epidermal growth factor receptor（EGFR）expression and alleviating airway injury in asthmatic rats and to reveal the underlying mechanism. MethodSD male rats were randomly divided into normal group, model group, dexamethasone group (5×10-4 g·kg-1) and Maxing Shigantang 1∶0.5, 1∶1, 1∶2 groups (group A, B, C, 10 g·kg-1), with 8 rats in each group. The other groups except the normal group received nebulization of 2% acetylcholine chloride and 0.4% histamine phosphate for the modeling of asthma. One hour before modeling， the normal group and the model group were given the same amount of normal saline， and the other groups were given the same amount of corresponding drugs， once a day for 7 days. On the 7th day， the model was established and the incubation period of asthma was recorded. The rats were then immediately anesthetized， and arterial blood and tracheal tissue were collected. Enzyme-linked immunosorbent assay （ELISA） was employed to detect the levels of interleukin-2 （IL-2）， interleukin-4 （IL-4）， and tumor necrosis factor-α （TNF-α） in serum. Pathological sections were prepared for the observation of the pathological changes of tracheal tissues and the ultrastructure of epithelial cells in each group. Terminal-deoxynucleotidyl transferase-mediated nick-end labeling （TUNEL） was adopted to detect epithelial cell apoptosis， and in situ hybridization and Western blot were employed to determine the mRNA and protein levels of epidermal growth factor receptor （EGFR）， respectively. ResultCompared with the model group， groups A， B and C prolonged the incubation period of asthma （P<0.05，P<0.01）. Compared with the control group， the model group showed declined IL-2 level （P<0.01）， risen IL-4 and TNF-α levels （P<0.05，P<0.01）， increased airway pathology score， collagen volume fraction， and airway epithelial cell apoptosis index （P<0.01）， and up-regulated mRNA and protein levels of EGFR in trachea tissue （P<0.01）. Compared with the model group， group A showed increased IL-2 level （P<0.05） and declined IL-4 （P<0.05，P<0.01） level， and group B showed declined IL-4 level （P<0.05）. The level of TNF-α in groups A， B， and C declined compared with that in the model group （P<0.01）. Maxing Shigantang repaired the tracheal tissue to different degrees （P<0.05）. Among the three groups， group A inhibited tracheal fibrosis （P<0.05） and had the most significant effect of repairing the ultrastructural changes of airway epithelial cells. Groups A， B and C all inhibited the apoptosis of airway epithelial cells （P<0.05）. All the three groups inhibited the up-regulation of EGFR mRNA level （P<0.05，P<0.01）， and groups B and C inhibited the up-regulation of EGFR protein level （P<0.05，P<0.01）. ConclusionMaxing Shigantang can inhibit the abnormal changes of airway epithelial structure， alleviate airway injury， and can down-regulate the expression of EGFR in the tracheal tissue of asthma model rats. In this study， the optimal compatibility of Maxing Shigantang to repair airway epithelial injury in asthmatic rats was group A， with the Ephedrae Herba-Armeniacae Semen Amarum-Glycyrrhizae Radix et Rhizoma-Gypsum Fibrosum ratio of 1∶0.5∶4∶1.
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AIM: To explore the expression and significance of forkhead box class O3(FOXO3)and interleukin-2(IL-2)in conjunctival epithelial cells and tears of patients with dry eye(DE).METHODS:A perspective study. A total of 106 DE patients who accepted from March 2019 to March 2021 were prospectively gathered, and 85 healthy subjects in the same period were selected as the control group. The level of FOXO3 in the conjunctival epithelial cells and tear fluid was measured by real-time fluorescent quantitative PCR(qRT-PCR)method; The level of IL-2 in the sample was measured by enzyme-linked immunosorbent(ELISA)method; The changes in clinical indicators of the ocular surface such as break-up time(BUT), Schirmer Ⅰtest(SⅠt), cornea fluorescein staining(CFS)in DE patients before and after treatment were analyzed; The correlation between the levels of FOXO3 and IL-2 in the conjunctival epithelial cells and tears of DE patients and the relationship between the two and clinical indicators were analyzed by Pearson correlation analysis.RESULTS:Compared with the control group, the level of FOXO3 in conjunctival epithelial cells and tear fluid in the DE group was obviously reduced, and the level of IL-2 was obviously increased(all P<0.01). Compared with before treatment, the level of FOXO3 in conjunctival epithelial cells and tear fluid of DE patients was obviously up-regulated, and the level of IL-2 was obviously down-regulated(all P<0.05). Pearson correlation analysis showed that the levels of FOXO3 and IL-2 in conjunctival epithelial cells and tear fluid were obviously inversely correlated(r=-0.531, -0.469, all P<0.01). After treatment, BUT and SⅠt indexes of DE patients increased compared with before treatment, while CFS decreased(all P<0.01). The level of FOXO3 in conjunctival epithelial cells of DE patients was obviously directly correlated with BUT and SⅠt(r=0.431, 0.457, all P<0.01), and it was obviously inversely correlated with CFS(r=-0.469, P<0.01), and the level of IL-2 was obviously inversely correlated with BUT and SⅠt(r=-0.416, -0.447, all P<0.01), and it was obviously directly correlated with CFS(r=0.424, P<0.01); tear FOXO3 was positively correlated with BUT and SⅠt(r=0.421, 0.443, all P<0.01), and it was negatively correlated with CFS(r=-0.474, P<0.01), and IL-2 was negatively correlated with BUT and SⅠt(r=-0.408, -0.429, all P<0.01), and it was positively correlated with CFS(r=0.419, P<0.01).CONCLUSION: the level of FOXO3 in conjunctival epithelial cells and tears of DE patients is decreased, and the level of IL-2 is increased. The two of which are closely related to the ocular surface indicators of patients. They are expected to become laboratory auxiliary indicators for clinical monitoring and prognostic evaluation of DE.
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Objective:To explore effects of different extracts and monomers of <italic>Lepidium meyenii </italic>（Maca） on the proliferation of mouse splenic lymphocytes and induction of interleukin-2 （IL-2） and tumor necrosis factor-<italic>α</italic> （TNF-<italic>α</italic>） by observing their immunomodulatory effects. Method:An octadecylsilyl （ODS） column was used to enrich the methanol extract of <italic>L. meyenii</italic> in stages to obtain six fractions and three monomers. Different groups of extracts and monomers of <italic>L. meyenii </italic>at different doses were set up. Cell counting Kit-8 （CCK-8） was used to detect the effect on the proliferation of mitogen-free， concanavalin A （Con A）-induced， and lipopolysaccharides （LPS）-induced mouse splenic lymphocytes. Enzyme-linked immunosorbent assay （ELISA） was used to determine the levels of IL-2 and TNF-<italic>α</italic>. Result:<italic>L. meyenii </italic>extracts Fr<sub>3</sub> and Fr<sub>6</sub>， and monomers <italic>N</italic>-benzyl hexadecanamide and 1，2-dihydro-4-carboxaldehyde-3-benzyl-<italic>N</italic>-hydroxypyridine slightly promoted the proliferation of Con A-induced T lymphocytes and LPS-induced B lymphocytes （<italic>P</italic><0.01） as compared with the conditions in the model group. <italic>L. meyenii</italic> extracts and monomers significantly induced the secretion of IL-2 and TNF-<italic>α</italic> by splenic lymphocytes （<italic>P</italic><0.01）. Conclusion:<italic>L. meyenii</italic> extracts and monomers can achieve immunological enhancement by promoting the secretion of IL-2 and TNF-<italic>α</italic>， and facilitate the proliferation of splenic lymphocytes. The active components are presumedly macamides and pyridine alkaloids， and the specific mechanism still needs to be further explored.
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Objective To study the effect of Cistanche deserticola Y C Ma (CDPS) on lymphocyte proliferation in mice. Methods The lymphocyte proliferation with or without mitogen was assessed by MTT assay in vitro. The immunosuppressed mice were induced by cyclophosphamide,and the spleen and thymus were weighted to determine the immune organ indexes in the normal or immunosuppressed mice. Thymocyte proliferation was employed to assess the activity of IL-2. Results CDPS significantly promoted both non-activated splenic lymphocytes and lymphocytes activated by ConA or LPS,and CDPS increased the secretion of IL-2 by splenic lymphocytes. CDPS (ip) remarkably increased indexes of spleen in normal or immunosuppressed mice,and also improved the indexes of immunosuppressed mice induced by cyclophosphamide. Conclusion CDPS can significantly promote the proliferation of splenic lymphocytes,and it may be related with promotion of secretion of IL-2 by splenocytes.
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Inflammatory bowel disease is thought to be regulated by the balance between Th1 and Th2 cytokines secreted by T cells, and NF-κB p65 also plays a predominant role in the intestinal inflammation. We evaluated the potency of oxymatrine, one of active components of Sophora Root, in inhibiting the immune responses and inflammation in 2,4,6-trinitrobenzene sulfonic acid(TNBS)-induced colitis. The inflammation was markedly ameliorated in the oxymatrine-treated rats.The level of IL-2 was increased and that of IL-10 was decreased in colon tissue in the rat model,which was reversed by the treatment of oxymatrine. Moreover, the elevated expression of NF-κB p65in colon tissue in the model was also improved by oxymatrine treatment. Our results suggest that oxymatrine might be beneficial for the abnormal immune responses and inflammation by regulating the unbalance of Th1 and Th2 cytokines secretion and inhibiting the expression of NF-κB p65 in colon tissue.
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[Objective]The study focus on the analgesic effect of polysaccharopeptide(PSP)and discuss the mechanism of this analgesia. [Methods]A total of 80 rats were divided randomly into high dose of psp,medium dose of psp,low doses of psp,positive control group,negative control group.After intragastric(ig) administration of different(high,medium and low)doses of PSP for 6 days,The tail stimulation-vocalization test was used to observe the analgesic effect of PSP.The IL-2 and IL-2R expression in mediobasal hypothalamus(MBH) was studied immunohistochemically.[Results]After intragastric(ig) administration of PSP for 6 days,the pain threshold was increased significantly and a dose-effect relationship was observed.After PSP administration the IL-2 expression in MBH was increased,while the IL-2R expression in MBH was decreased.[Conclusion]PSP has a definite analgesic effect and its analgesic site is in the MBH.The analgesia might be produced by the binding with the IL-2R in MBH by IL-2 secreted by T cells after PSP was injected into the brain.
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Objective: To prepare and identify recombinant human IL 2/GM CSF(rhIL 2/GM CSF) fusion protein antibodies and to study its specificity and its effect on fusion protein biological activity. Methods: rhIL 2 /GM CSF fusion protein was purified by DEAE Sepharose FF ion exchange chromatography. The purified protein was used to immunize rabbits for the preparation of antisera. The titer and specificity of the antisera were detected by ELISA and Dot ELISA and the biological activity by cell proliferation. Results: The antisera not only reacted with the rhIL 2/GM CSF, IL 2 and GM CSF, but also inhibited the biological activity of the rhIL 2/GM CSF, IL 2 and GM CSF. Conclusion: The obtained antisera can be used to study the structure and function of the rhIL 2/GM CSF.
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The in vitro modulation effect of insulin on DNA and IL-2 production of spleen lymphocytes in diabetic mice were studied. The results suggest that the DNA and IL-2 production of the lymphoc-ytes are significantly inhibited in the mice. When suspend the lymphocytes to the culture medium containing insulin, the DNA and IL -2 production of the lymphocytes are remarkably increased. Therefore, insulin is an important immunoregulation hormone.
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The level of serum soluble interleukin 2 receptor(sIL-2R)was studied in 29chronic persistent hepatitis (CPH) patients and in 18 normal individuals.It was found that sIL-2R level was significantly higher in the CPH patients than that in normal individuals(P
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Es inducing human spleen cells in 30 cases were studied. We found that Es could not only induce IFN-? from human spleen cells, but also IL-2 and LT. The mixed lymphokines containing IFN-?(3269.6?1634.4 U/ml), IL-2 (64.8?40.8 U/ml) and LT (54.4?44.6 U/ml) were induced by ES. Our study shows that the mixed lymphokines appeared to be cytotoxic to human SMMC-7721, SPC-3, HcLa, Jutkat arid Molt-4 cells lines at all levels, but it had no cytotoxic effect to normal cells (WISH cells).
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Objective:To study the effect of pig interleukin 2(IL 2) eukaryon expression plasmid on cellular immune responses of BALB/c mice immuned with pcDNA PRRSV ORF5 DNA vaccine.Methods:BALB/c mice were immunized with pcDNA PRRSV ORF5 DNA vaccine and pig interleukin 2(IL 2) eukaryon expression plasmid by the routes of co injection and DNA vaccine injection alone respectively, with PBS and pcDNA3 1(+) as controls. Fluoresecence Activated Cell Sorter(FACS),T lymphocyte proliferation test(MTT) were used to detect the number of CD4 +、CD8 + and the T lymphocyte proliferation in peripheral blood of mice vaccinated.Results:ConA response of T lymphocytes in blood was higher in experiment group than the control group ( P
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Objective To evaluate the effect of ultra-filtration extract from Danggui Buxue Decoction (DBD) on immune function of H22 bearing mouse and explore the mechanism through observing the effect of ultra-filtration extract from DBD on the secretion of the cytokines (IL-2,IFN-?).Methods We used membrane separation technique to extract DBD.The content of IL-2 and IFN-? in splenocyte culture supernatant was detected by enzyme-linked immunosorbent assay (ELISA),and the mRNA expression of cytokines (IL-2,IFN-?) in splenocytes was assayed by reverse transcription-polymerase chain reaction ( RT-PCR).Results The content of IL-2 and IFN-? in splenocyte culture supernatant was increased in ultra-filtration extract from DBD groups,significantly in high-dose group (P
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Objective To investigate the relationship between in vivo anti-tumor efficacy of Shenqi injection and its effect on immunologic function in H22 tumor-bearing model mice,and explore the cellular immunologic mechanism of Shenqi injection in inhibiting tumor growth.Methods Three different doses of Shenqi injection were given to H22 tumor-bearing model mice in treatment groups.The tumor growth inhibitory rate(IR) and the index of phagocytosis of peritoneal macrophage were calculated.MTT assay was used to observe the effect of Shenqi injection on spleen lymphocytes stimulated by ConA in vitro separated from H22 tumor-bearing mice.Murine serum IL-2 and IFN-? were detected by means of ELISA assay.Results IR was significantly reduced in two treatment groups compared with that of the model mice(60.72%,48.65%,P
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In order to determine the cellular immunestate in patients with multiple myeloma, T-cellsubsets in peripheral blood from 25 cases withmultiple myeloma were measured with SPA-Ig di-rect rosette assay, the activity of interleukin-2 wasinvestigated by using mice thymocyte proliferationassay. The results showed that in multiple myelo-ma, OKT_3~+, OKT_4~+ cell decreased significantly.OKT_6~+ cell increased markedly and the OKT_4/OKT_8 ratio was reduced. The activity of IL-2 in pe-ripheral blood lymphocytes of the patients withmultiple myeloma increased sharply and there wasa significant difference between the active diseaseand the remission disease. The activity of IL-2 inthe remission disease was lower than that in theactive disease but higer than that of the controlgroup. In conclusion, this study indicated thatthere were imbalance of T-cell subsets and abnor-mality of IL-2 production in patients with multiplemyeloma. The data might suggest that there wasdisorder of the cellular immunity in the course ofthe disease.
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In this paper, murine natural killer (NK) cells were studied with cytochemistry and electron-microscopy after treated with interleukin-2 (IL-2) and polysaccharide of astragalus (PA). The results showed that acid phosphatase (AcP), acid naphthylacetate esterase (ANAE) and naphthol-AS-D chloroacetate esterase (NCAE) were positive in NK cells, and the activity of AcP and ANAE was enhanced by IL-2 and PA. Significant changes of NK cells in ultrastructure took place after preculturing with IL-2.