| Objective:(1) To investigate the therapeutic effect of Chinese medicine Eczema prescription No.1 on acute dermatitis and eczema in mice and explore the mechanisma;(2) To explore its regulatory mechanism on T-lymphocyte subsets in mice by studying the effect of Chinese Medicine Eczema Prescription No.1 on splenic T-lymphocyte subsets in mice;(3) To research that if Chinese medicine Eczema prescription No.1 can down-regulate the NF-KB signaling pathway to inhibit eczema by studying the effect of Eczema Prescription No.1 on the nucleus NF-KB p65, NF-KB1 p50 protein expression.Methods:(1) Make acute eczema models 2,4-dinitrofluorobenzene(DNFB) wa s used for sensitizing the skin on 50 BALB/c mice's abdomens and ears to m ake acute eczema models. The 50 modelled mice were randomly and evenly di vided into 5 groups-- control group treated with normal saline, positive control group with hydrocortisone, and the other three experimental groups with low,medium or high dosage of Eczema prescription 1;(2) Observe the parameters We used a puncher to take tissues with same size from middle part of mic e's bilateral ears, and weighed the tissues. The degree of swelling was defined as the difference between the weight of bilateral ears. Another part of the tiss ues were HE stained, and observed the histopathologic change under an electro nic microscope;(3) The detection of cytokines The mice's blood was collecte d from the inner canthus before the treatment, and through removing eyeballs a fter the treatment; After serum separation, we detected the expression of IL-2?IL-1??IFN-??IL-4?IL-5 and TNF-? in peripheral blood with the enzyme link ed immunosorbent assay(ELISA);After the last administration, the spleens wer e took out and the lymphocytes were extracted in sterile conditions. CD3, CD4,CD45 were measured by flow cytometry;.(4) Determination of NF-KB The samples were taken from the left ears of mice, and the expression of NF-KB p65 and NF-KB1 p50 in the nucleus was determined by Western-blot method.Results:(1) In the groups of various dosages of Eczema prescription No.1 as well as in the positive control group, the decreases of ear swelling degree were great, which was significantly different from the control group(P < 0.05). The high-dosage group and the positive control group group were much better than the medium- dosage group and the low-dosage group(P < 0.05); From pathology, compared with the control group, the improvements in intercellular edema, intracellular edema and lymphocyte infiltration of the high-dose group and the medium-dose group were no less than those of positive control group;(2)After modelling, compared with the normal group,the serum IL-2?IFN-??IL-1??TNF-? increasedand IL-4 decreased significantly(P < 0.05). There was no significant difference in the above factors between each group(P > 0.05); But, the level of IL-5 didn't change obviouslyafter modelling. After treatment, compared with the NS group, the groups of various dose of Eczema prescription No.1 and the hydrocortisone groupcould significantly lower the levels of IL-2(P < 0.05)and increase the level of IL-4(P < 0.05); In the high-dose group, the medium-dose group and the hydrocortisone group, IFN-? and TNF-? are much lower than those in the NS group(P < 0.05). And the effects on the expression of IFN-? in the high-dose group and the hydrocortisone group were statisticallylower than that in the low-dose group(P < 0.05); In addition, the hydrocortisone group was also superior tothe low-dose groupon the improvement of cytokine TNF-?(P < 0.05);But, besides the NS group,other groupshad no significant effect on the level of serum IL-1??IL-5( P > 0.05).(3) The results showed that the ratios of the CD3 and CD4T-lymphocytes in the high dose and in the hydrocortisone group raised obviously than those of the control group(P < 0.05),hoever,compared to the hydrocortisone group group, the hydrocortisone group group was superior to the the high dose(P < 0.05).There were high expressions of CD45 antigen in all groups, but no significant difference were found in the ratios of CD45 positive cells between the groups.(4) The expressions of NF-KB1 p50 were much less in the high-dose group than those in the hydrocortisone group,the low-dose group, the medium-dose group and the NS group(P < 0.05). And the higher is the Materia Medica dosage, the lower is the expression?Conclusions:(1) In the mice with acute eczema, Eczema prescription No.1 ca n actively suppress ear swelling and decrease cellular infiltration in the skin le sions.It may suppress mice's eczema through inhibiting the release of IL-2, IF N-?and TNF-?, and raising the expression of IL-4.It's probably irrelevant to the level of IL-1? and IL-5.(2) High dose of Eczema prescription No.1 can signif icantly increase the ratios of CD3 and CD4 T-lymphocytes, which indicates tha t it may enhance the immune function by increasing the signal transduction of T cells.(3)Eczema prescription No.1 in various dose can down-regulate the e xpression of NF-KB1 p50. And the higher is the dose, the lower is the expres sion. This may be one of the main mechanisms in the anti-inflammation. |
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