| Objective:in this study,a rat model of chronic eczema was established to observe the effects of Shi Zhen Qing granules on skin damage degree,inflammation,immunity and HPA axis in rats with chronic eczema,and to preliminarily explore the mechanism of Shi Zhen Qing granules.It provides relevant theoretical and experimental basis for the promotion and use of Shi Zhen Qing granules in clinical practice,and provides data reference for the further research on the mechanism of action of Shi Zhen Qing granules.Methods:60 rats were randomly divided into 6 groups: blank group,model group,positive control group,and Shi Zhen Qing granule high,medium and low dose groups.Except the blank group,7% dinitrochlorobenzene(DNCB)acetone solution was applied to the rat model of chronic eczema.After successful modeling,the drug was given for 7 days according to body weight,and weighed 12 hours after the last administration.Blood was taken from the abdominal aorta after ether anesthesia,and the spleen,thymus and affected skin were removed.Abdominal aorta blood was collected,centrifuged at 2500r/min for20 min,serum was separated,serum was stored in the cryo-tube and stored in the refrigerator at-80 ?.After weighing the spleen and thymus directly,the viscera index was calculated.The affected skin tissue was fixed in 10% formaldehyde solution,and the pathological section was made.HE staining was used to observe the histopathological changes of local chronic eczema skin lesions in rats.Serum levels of inflammatory cytokines IL-4,IL-25,IL-31,IFN-?,COTR,ACTH and CRH were determined by enzyme-linked immunosorbent assay.The data of each index were statistically analyzed, and the software SPSS22.0 was used for one-way analysis of variance.Results:1.The chronic eczema model of rats was induced by DNCB,so that the skin on the back of the rats showed changes of erythema,papules,scales,lichen,and other chronic eczema like skin lesions.After the model was built,the animals kept scratching and rolling,which was consistent with the successful performance of the model of chronic eczema in experimental zoology.The score of skin lesions in the blank group was 0(total 3),and that in the model group was 2.74,2.62,2.63,2.62 and 2.63,respectively,showing clinical statistical significance(P < 0.05).The rat model of chronic eczema was successfully replicated.2.HE staining results indicate model visible skin of pathological changes of rats,and extend the prickle cell layer thickening,epidermis in see a large number of inflammatory cells infiltration,visible strips neutrophil infiltration,subcutaneous blood capillary rise,vasodilation,histological,conforms to the skin manifestations and pathological changes of chronic eczema,further supporting the rat model with chronic eczema copy success of the project group.By prednisone,Shi Zhen Qing particles high,medium and low dose prednisone treatment for 7 days,Shi Zhen Qing particles high,medium and low dose group can effectively reduce the rat skin erythema,infiltration,scales,such as clinical symptoms,reduce lesion scores(P < 0.01),and can effectively suppress chronic eczema skin epidermis tu extend,hyperkeratosis and parakeratosis,thickening of stratum spinosum,reduce or eliminate the inflammatory cells infiltration,inhibit inflammation and other symptoms,and with chronic eczema model group with significant difference.3.The results of immune organ index showed that,compared with the blank group,the spleen index and thymus index in the chronic eczema model group were significantly lower(P < 0.01 or P < 0.05).Compared with the model group,the spleen index and thymus index decreased in the model of chronic eczema were increased in the positive control group and in the high,medium and low dose groups of Shi Zhen Qing granules(P < 0.01 or P < 0.05).Compared with the positive group,the spleen index increased in the high and medium dose groups of Shi Zhen Qing granules(P<0.01),while the thymus index showed no difference(P>0.05).The results showed that Shi Zhen Qing granule group could significantly improve the spleen index and thymus index of the model of chronic eczema,and improve the immune organ function of the model rats,and the effect of Shi Zhen Qing granule in the high-dose and medium-dose group was better than that in the positive control group.4.ELISA results showed that compared with the blank group,CRH,ACTH and CORT in serum of rats with chronic eczema were abnormal and their contents were significantly reduced(P<0.01).Compared with the model group of chronic eczema,the contents of CRH,ACTH and CORT in serum of the model of chronic eczema were significantly increased in the high,medium and low dose Shi Zhen Qing granules group and the positive control group(P<0.01 or P<0.05).Compared with each treatment group,the contents of CRH,ACTH and CORT in serum of rats in the high-dose group of Shi Zhen Qing granules were significantly higher than those in the positive control group(P < 0.01),while the contents of CRH and CORT in serum of rats in the medium-dose group of Shi Zhen Qing granules were significantly higher than those in the positive control group,while the contents of ACTH were lower than those in the positive control group(P < 0.01).The content of CRH,ACTH and CORT in the Shi Zhen Qing low-dose group was significantly lower than that in the positive control group,but higher than that in the model group(P<0.01 or P<0.05).The contents of CRH,ACTH and CORT in the serum of rats were higher in the high-dose group than in the medium-dose group,and higher in the medium-dose group than in the low-dose group,with statistically significant differences(P<0.01).The results showed that the contents of CRH,ACTH and CORT in serum of the rats with chronic eczema caused by DNCB were lower than those in the normal group,indicating that the HPA axis function of the rats with chronic eczema model was low,suggesting that the pathogenesis of chronic eczema may have low HPA axis function.Shi Zhen Qing granules can effectively increase the content of CRH,ACTH and CORT in the serum of chronic eczema model,stimulate the function of HPA axis,and promote the expression of glucocorticoid,thus suggesting that Shi Zhen Qing granules can inhibit immune inflammation.The effect of Shi Zhen Qing granule in high and medium dose group was better than that in positive control group.Compared with the blank group,the expression of IFN-? in serum of rats with chronic eczema decreased,while the expression of IL-4,IL-25 and IL-31 increased(P<0.01).Compared with the model group,the serum levels of IFN-? in the positive control group and the high-dose,medium-dose and low-dose groups of Shi Zhen Qing particles were significantly increased,while the expressions of IL-4,IL-25 and IL-31 were decreased(P<0.01 or P<0.05).It was found that the overall effect of Shi Zhen Qing granules in the high and medium dose groups was significantly better than that in the positive control group(P<0.05).Results show that the Shi Zhen Qing particles high,medium and low dose group can effectively reduce the chronic eczema caused by model has increased in rats serum IL-4,IL-25,IL-31 content,increase of IFN-? content in serum of rats,so as to adjust the Th1 / Th2 immune imbalance expression to relieve chronic eczema,allergic reaction,reduce inflammatory symptoms,itching,so as to relieve the clinical symptoms of chronic eczema.Conclusions:1.Shi Zhen Qing granules can effectively alleviate the degree of skin damage in rats with chronic eczema,reduce the score of skin damage,reduce the degree of inflammation and itching,delay the development of the disease in rats with chronic eczema,and effectively control the development of the disease.2.Shi Zhen Qing granules can effectively increase the decreased levels of CRH,ACTH and CORT in the serum of rats with chronic eczema,and its mechanism of action may promote the expression of glucocorticoid by stimulating the function of HPA axis,thus inhibiting the inflammatory expression of eczema.3.Shi Zhen Qing granules can significantly improve the decreased spleen index and thymus index in the model of chronic eczema,and improve the immune organ function in the model of chronic eczema rats,indicating that the mechanism of action may be related to the regulation of immune function.4.Shi Zhen Qing granules can effectively reduce the contents of inflammatory factors IL-4,IL-25 and IL-31 in the serum of rats with chronic eczema model due to overexpression,and increase the content of anti-inflammatory factor IFN-? in the serum of rats,so as to relieve inflammatory symptoms and itch expression,and thus reduce the clinical symptoms of chronic eczema.The mechanism is related to the regulation of the expression of Th1 and Th2 cytokines. |
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