| Allergic disease,a chronic inflammatory disease,is the abnormal immune response of the body happened when antigens stimulate the body contributing to the tissue injury or dysfunction.It includes allergic asthma,allergic rhinitis,and allergic dermatitis etc.Allergic diseases have three characteristics:rapid onset,intense reaction and fast regression.When body exposed to the antigen again,the reaction will be more rapid and strong,then the second disease occurs.Its pathogenesis is complicated and has not yet been fully clarified.Nowadays,there is a new view that allergic diseases may be transmitted from generation to generation.Therefore,how to control the process of allergic diseases,improve the allergic physique of the offspring,and alleviate the recurrence of allergic diseases still need to be explored.Based on the precious study,this paper mainly focuses on the following aspects:1.To explore whether maternal allergic diseases may take effect on offspring.Murine maternal allergic contact dermatitis(ACD)-offspring ACD model and murine ACD 1)before or 2)after maternal-offspring ACD model and 3)murine maternal FITC-sensitive model were established to investigate the effect on offspring’s ear swelling,ear histopathology,and Enzyme-Linked ImmunoSorbnent Assay(ELISA)to detect the expression levels of type II cytokines such as:IL-4,IL-5,and IL-13 in the supernatant of right ear homogenate,flow cytometry to detect ILC2s in mouse peripheral blood mononuclear cells(PBMC),cervical lymph nodes,and axillary lymph node cells.2.Explore the possible mechanism of transmission of allergic diseases.1)Effects of cytokines and their receptors’ expression in the embryos and placenta of ACD maternal:Murine maternal FITC-induced allergic dermatitis model was established.Maternal ear thickness was measured 24 hours after challenge,and the degree of ear swelling was calculated.At the 19th day of pregnancy,embryos and placenta were taken out,and mononuclear cells were extracted for flow cytometry.The expression levels of cytokines IL-4,IL-5,IL-13,IL-33 in embryos and placenta were detected by ELISA.The expression of mRNA:il13rα1,il13rα2,il17ra,il17rc,and tslpr2 in embryonic tissues were detected by qRT-PCR.2)Changes in the number of ILC2 and Th2 cells in different tissues of the offspringof ACD maternal:Murine maternal FITC-induced allergic dermatitis model were established,flow cytometry was used to detect mononuclear cells in the aervical lymph nodes,axillary lymph nodes,mediastinal lymph nodes,mesenteric lymph nodes,small intestine and PBMCs.Murine maternal FITC-sensitive model was established flow cytometry was used to detect mononuclear cells in the aervical lymph nodes,amediastinal lymph nodes and mesenteric lymph node.3)The effect of IL-4 on ILC2 in PBMC in vitro:extract mouse PBMC,add 100ng/ml IL-4 to stimulate in vitro for 24h,use flow cytometry to detect the amount of Lin-KLRG1+IL-25R+ILC2.4)Verification of the role of IL-4 and IL-13 in the intergenerational transmission of allergic dermatitis:Murine maternal FITC-induced allergic dermatitis model was established,the maternal were injected with IL-4 or IL-13 neutralizing antibodies after challenge.Offspring FITC-induced allergic dermatitis model were established at the age of four weeks after birth,and the ear thickness was measured for 24 hours with an interval of every 2 hours after the challenge.The degree of ear swelling was calculated,the pathological changes of the ear tissue were observed,and the expression levels of type II cytokines IL-4,IL-5,and IL-13 in the supernatant of right ear homogenate were detected by ELISA.ILC2s in offspring peripheral blood mononuclear cells(PBMC),cervical lymph nodes,and axillary lymph nodes were detected by flow cytometry.3.To explore whether the administration of Yu-ping-feng-san in murine ACD recurrence model may alleviate the allergic symptoms of the offspring.Murine maternal FITC-induced allergic dermatitis model,and offspring FITC-induced allergic dermatitis model were established when offspring develop allergic diseases in childhood how to relieve symptoms.After the first time after challenge,when the ear swelling is completely relieved,offspring were administrated with YPFS 10 days on remission.After the administration,then we elicited the inflammation again 24 hours after the last administration.Ofspring’s ear swelling,ear histopathology was detected in 12 huors with an interval of every 2 hours,the expression levels of type Ⅱ cytokines IL-4,IL-5,and IL-13 in the supernatant of right ear homogenate were detected by ELISA and ILC2s in offspring PBMC were detected by flow cytometry.The results showed that:1.Maternal’s allergic disease will increase the susceptibility of offspring.Allergic diseases occurred in the mothers,and the allergic symptoms were more severe when the offspring developed allergic diseases,the time of allergic diseases were lasted longer,and had an earlier onset;the ear tissues of the ACD offspring were significantly thickened,and the pathological results showed that the maternal ACD-offspring ACD’s ear tissue thickened were more obviously and accompanied with edema.At the same time,the inflammatory cell infiltration was more obvious,and the ear tissue dermis tissue was obviously edema.The results of type II cytokines’ expression level also showed that the level of IL-13 in the ear tissue homogenate of the ACD offspring from ACD mother were significantly higher than that of the ACD offspring of control mother.The flow cytometry results did not show significant changes.The results indicate that if mothers develop allergic diseases,their ACD offspring will develop the allergic disease earlier when the onset time is longer,the duration is longer,and the severity of the disease is more serious.This result suggests that the allergic disease of mother will increase offspring’s susceptibility.However,in murine maternal FITC-sensitive model,the offspring’s ear swelling degree,ear weight,type II cytokines’ expression,ear tissue pathology compared with the control mother’s FITC offspring showed no significant difference.The above results suggest that maternal experience a complete allergic dermatitis process,the onset time of allergic dermatitis in the offspring of ACD was earlier,the duration was longer,and the severity of the onset was more serious,indicating that there was intergenerational transmission of allergic dermatitis.However,sensitization by contact with allergens alone cannot cause changes in the susceptibility of the offspring.2.Allergic diseases transmission from maternal to offspring is related to IL-4 and IL-13.1)Effects of cytokines and t.heir receptors’expression in the embryos and placenta of ACD maternal:The mononuclear cells in the embryo and placenta were detected by flow cytometry,and no significant changes were found.The detection of the expression level of type II cytokines in embryos and placenta showed that IL-4 and IL-13 in embryos and placenta of the model group were significantly higher than those in the control group.Detection of mRNA:il4rα,ill3rα1,il13rα2,ill 7ra,ill7rc,tslpr2 in embryos showed that compared with the control group,the expression of il4rα in the embryos of the model group had a certain upward trend.It is suggested that the allergic disease of the mother may affect the offspring through IL-4 and IL-13.2)Change.s in the number of ILC2 and Th2 cells in different tissues of the_offspring of ACD maternal:The mononuclear cells in different parts were detected by flow cytometry,the results showed that:in murine maternal FITC-induced allergic dermatitis model,Lin’KLRG1+IL-25R+ILC2 in the amediastinal lymph nodes and mesenteric lymph nodes were increased significantly,but there was no significant difference in all parts of the offspring of murine maternal FITC-sensitive model.This result suggests that the intergenerational transmission of allergic dermatitis may be related to Lin-KLRG1+IL-25R+ILC2.However,in murine maternal FITC-induced allergic dermatitis model ILC2 in the amediastinal lymph nodes and mesenteric lymph node showed no significant difference.3)The effect of IL-4 stimulation on ILC2 in vitro:After IL-4 stimulated PBMC in vitro for 24 hours,the number of Lin-KLRG1+IL-25R+ILC2 in the IL-4 group increased significantly compared with the control group.This result suggests that the IL-4 produced by the mother’s allergic dermatitis can directly affect the number of ILC2 in the offspring.4)Verification of the role of IL-4 and IL-13 in the intergenerational transmission of allergic dermatitis:Establish murine maternal FITC-induced allergic dermatitis model,at the same time of maternal were challenged,injected intraperitoneally with 100 μg IL-4 and IL-13 neutralizing antibodies.The offspring established a FITC-induced allergic dermatitis model.The results showed that after when mothers were-injected with IL-4 and IL13 neutralizing antibodies,ear inflammation including ear swelling and inflammatory cell infiltration of the offspring,which develops allergic diseases again,were significantly relieved.At the same time,the expression levels of type Ⅱ cytokines IL-4、IL-5 and IL-13 in the supernatant of ear tissue homogenate were decreased.This result suggests that the susceptibility of the offspring is increased through IL-4 and IL-13 due to the allergic disease in the mother,while the susceptibility of the offspring will be reduced after the neutralization’s administration of IL-4 and IL-13.3.YPFS administrated at the remission stage of the offspring of maternal ACD-offspring ACD model could inhibit allergic inflammation.YPFS(6.5 mg/kg)administration in remission for ten days not only had a significant inhibitory effect on the ear inflammation with ear swelling and inflammatory cell infiltration of ACD maternal’s ACD offspring,but also inhibit the expression levels of type II cytokines IL-4、IL-5 and IL-13 in the supernatant of ear tissue homogenate.This suggests that the administration of YPFS in the offspring’s remission period can exert an effect by inhibit allergic reactions.Conclusions:1.Maternal’s allergic dermatitis will increase the susceptibility of offspring.2.Allergic dermatitis transmission from maternal to offspring is related to IL-4、IL-13 and Lin-KLRG1+IL-25R+ILC2.3.The administration of YPFS during remission period of offspring ACD recurrence model can inhibit its allergic effect. |
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