银杏叶提取物对哮喘大鼠气道炎性细胞蛋白激酶Cα表达及白细胞介素-4分泌的影响

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目的探讨银杏叶提取物(EGb)对哮喘的治疗效果及可能机制。方法48只SD大鼠随机分成:正常对照组(6只)、哮喘组(此组再分为:未处理组,激素处理1、2、4周组,EGb处理1、2、4周组,每组各6只)。采用Wright染色法计数肺泡灌洗液中各类炎性细胞的相对数,免疫组化(SP)法测定蛋白激酶Cα(PKCα)的表达情况,ELISA法测定肺泡灌洗液上清中IL-4的含量。结果哮喘未处理组大鼠肺泡灌洗液中嗜酸性粒细胞(EOS)、淋巴细胞相对计数百分比、PKCα在淋巴细胞和总的细胞中的阳性表达率以及上清液中IL-4含量均显著高于正常对照组(均P<0.05)。与未处理组比较,激素处理不同时间组前述各指标均明显下降(均P<0.05)。EGb处理1、2周组各指标明显高于激素处理组(P<0.05),4周组与激素处理组比较无明显差异。EGb处理1周组除淋巴细胞PKCα阳性表达率外,其余检测指标与哮喘未处理组比较均无明显差异(均P>0.05),而EGb处理2、4周组相关检测指标均明显下降(均P<0.05),且随时间延长,下降更明显(P<0.05)。EGb处理组炎性细胞PKCα阳性表达率与上清液中IL-4、嗜酸性粒细胞及淋巴细胞的相对计数百分比呈显著正相关(r值分别为0.641、0.699、0.625,均P<0.01,n=18)。结论EGb可减轻气道EOS和淋巴细胞等炎性细胞的浸润以及气道炎性细胞PKCα的阳性表达率和IL-4的分泌量,其作用弱于糖皮质激素,但随着药物治疗时间的延长其药理作用逐渐明显。EGb治疗哮喘的作用可能与其减少炎性细胞PKCα的表达进而降低EOS和淋巴细胞在气道的浸润以及IL-4的浓度有关。 Objective To explore the therapeutic effect and possible mechanism of Ginkgo biloba extract (EGb) on asthma. Methods Forty-eight Sprague-Dawley rats were randomly divided into normal control group (6 rats) and asthma group (this group was subdivided into untreated group, hormone treatment group 1, 2, 4 weeks group, EGb treatment group 1, 2, 4 weeks group. 6 in each group). The relative numbers of inflammatory cells in bronchoalveolar lavage fluid were counted by Wright staining, and the expression of protein kinase Cα (PKCα) was measured by immunohistochemistry (SP) method. The IL-4 in the supernatant of bronchoalveolar lavage fluid was measured by ELISA. The content. Results The percentage of eosinophils (EOS), the relative percentage of lymphocytes in bronchoalveolar lavage fluid, the positive rate of PKCα in lymphocytes and total cells, and the content of IL-4 in supernatants were significant in the asthma treated untreated group. Higher than the normal control group (all P < 0.05). Compared with the untreated group, the above-mentioned indexes in the hormone treatment groups at different time points were significantly decreased (all P<0.05). The indexes of EGb treatment in the 1st and 2nd week groups were significantly higher than those in the hormone treatment group (P<0.05). There was no significant difference between the 4 week group and the hormone treatment group. In addition to the PKCα positive expression rate of lymphocytes in the EGb treatment group for 1 week, there was no significant difference between the other indicators and the untreated asthma group (all P>0.05), while the relevant detection indexes of the EGb treatment group were significantly decreased in the 2nd and 4th week groups (both P<0.05), and the decline was more pronounced with time (P<0.05). The positive expression rate of PKCα in inflammatory cells of EGb treatment group was positively correlated with the relative counts of IL-4, eosinophils and lymphocytes in the supernatant (r values ​​were 0.641, 0.699, and 0.625, respectively, all P<0.01). n=18). Conclusion EGb can reduce the infiltration of inflammatory cells such as airway EOS and lymphocytes, as well as the positive expression rate of PKCα and IL-4 secretion in airway inflammatory cells. Its effect is weaker than that of glucocorticoid, but with the time of drug treatment. Extend its pharmacological effects gradually. The effect of EGb in the treatment of asthma may be related to the reduction of the expression of PKCα in inflammatory cells and thus the reduction of EOS and lymphocyte infiltration in the airways and the concentration of IL-4.
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