The Experimental Study On Xiaofeng Powder In The Inhibiting Function And The Immunology Mechanism To Allergy Contact Dermatitis

ObjectiveThe mouse model of allergy contact dermatitis was used in this study.From the pathological changes, associated immune cells, cytokine andcytokine mRNA expression, this in-depth research is carried through in thepharmacology effect and the immunology mechanism about that Xiaofengpowder control allergy contact dermatitis (anti- hypersensitivity). This study ofXiaofeng powder could further provide the laboratory basis on the wide spreadapplication in clinical. To study this prescription on the immune mechanism,we could also discuss the law of TCM understanding diseases and theorganism function. It can promote to the modernization of Chinese medicineskin theory.MethodsAt first the mouse model of allergic contact dermatitis was formed byusing DNCB. Through observed in mouse ears thickness, swelling, swellinginhibition percentage, indexes of spleen and thymus, there is a preliminaryevaluation for the roie of the Xiaofeng powder on anti-hypersensitivity. HEpathological examination and Transmission Electron Microscope (TEM) examination were performed to determine the effects of Xiaofeng Powder onultrastructure of skin and its curative effect to the ACD. Then using the bloodcorpuscle analyzer, leukocyte, neutrophilic granulocyte, lymphocyte andmonocytes were measured. Meanwhile, enzyme-linked immunosorbent assay(ELISA) was performed to examine the levels of interleukin-4 (IL-4),interferon-gamma (IFN-γ) and soluble interleukin-2 receptor (sIL-2R) in theears of these mice. To study the molecular mechanism of Xiaofeng powder onregulation of the Th1/Th2 cytokine network balance in the blood,immuno-histochemistry and RT-PCR technique were used to detect theexpression of IL-10,IL-13,IFN-γand IL-6 in the tissues.Results1. Xiaofeng powder reduced the auricle thickness and weight in the ACDmiceCompared with the NS group, the auricle thickness of ACD group mice isincreased. The difference was statistically significant (P＜0.05). At the sametime weight and the swelling of the auricle in the ACD group was found tosignificantly increased (P＜0.05). Compared with ACD group, the auriclethickness, weight and the swelling of the auricle in the Xiaofeng powder groupsignificantly reduced (P＜0.05).2. Xiaofeng powder reduced the indexes of spleen and thymus in the ACDmiceThe indexes of spleen and thymus of the ACD group were significantlyhigher compared with the NS group (P＜0.05). The indexes of spleen andthymus of the Xiaofeng powder and the prednisone group decreased. Thedifference was statistically significant (P＜0.05). There was no differencebetween the Xiaofeng powder and the prednisone group (P＞0.05).3. Xiaofeng powder eased the damage degree on pathological features andultrastructural features of the mouse derma in the ACD mice Pathological sections of skin tissues from the ACD group mousedisplayed that those large quantities of inflammation cells infiltrated in theepidermal basal layer and spine and interstitial edema. After administration ofXiaofeng powder, the skin of epidermal damage was reduced. The intercellularedema was obviously alleviated and inflammation cells in the dermis havedeclined markedly.Under TEM, the electron density of epidermal keratinocyte cellsincreased slightly. The spinous cells and basal cells appeared to wither and cellgaps widened. In cytoplasm cell organs showed vacuolar changes. Thechromatin in nuclear condensed into massive conglobation. The cristae ofmitochondrial fractured and were in vacuolar changes. After administration ofXiaofeng powder, the organizational structure of various layers was regulated.Changes in the spinous cells and cell gaps were significantly improved.Changes in basal cell gaps and the cell nucleus were obviously reduced. Incytoplasm there were structural integrity of the mitochondria and ribosome.4. Xiaofeng powder decrease leukocyte, neutrophilic granulocyte,lymphocyte counts in the blood of the ACD miceIn the ACD group leukocytes, neutrophilic granulocyte and lymphocytewere significantly higher than the NS group mice. The difference wassignificant (P＜0.05). Leukocytes, neutrophilic granulocyte and lymphocytewere declined in the Xiaofeng powder group and the prednisone group.Compared with the ACD group, the difference was significant (P＜0.05). Butthe Xiaofeng powder had little impacts on monocytes. Compared with the NSgroup there was no significant difference (P＞0.05).5. Xiaofeng powder upregulated the levels of IL-4, declined the levels ofIFN-γand sIL-2R in serum of the ACD miceThe levels of IL,4 in the Xiaofeng powder group and the prednisonegroup were higher than those in the ACD group (P＜0.05). The levels of IFN-γ and slL-2R in the Xiaofeng powder group and the prednisone group werelower than those in the ACD group (P＜0.05). But the levels of IL-4, IFN-γand sIL-2R were not significantly different between the Xiaofeng powdergroup and the prednisone group (P＞0.05).6. Xiaofeng powder upregulated the levels of IL-10 and declined thelevels of IL-13 in the mouse derma of the ACD miceIn the mouse derma of the NC group IL-10 was lacked. The expression ofIL-10 was low-level in the ACD group. Compared with the NS group therewas not significantly different (P＞0.05). The levels of IL-10 in the Xiaofengpowder group and the prednisone group were increased higher than those inthe ACD group (P＜0.05). In the mouse derma of the NC group IL-13 waslacked. The expression of IL-13 was higher in the ACD group. Compared withthe NS group there was significantly different (P＜0.05). The levels of IL-13 inthe Xiaofeng powder group and the prednisone group were decreased lowerthan those in the ACD group (P＜0.05).7. Xiaofeng powder decreased the expression of IFN-γmRNA, but had nosignificant effect on the expression of IL-6 mRNA in the mouse auricle of theACD miceIn mouse auricle cells of the ACD group the IFN-γmRNA expressionwas significantly increased higher than those in the NS group (P＜0.05). Thelevels of IFN-γmRNA in the Xiaofeng powder group and the prednisonegroup were decreased lower than those in the ACD group (P＜0.05). In mouseauricle cells of the ACD group the IL-6 mRNA expression was significantlyincreased higher than those in the NS group (P＜0.05). But the levels of IL-6mRNA in the Xiaofeng powder group and the prednisone group were notremarkable different than those in the ACD group (P＞0.05).8. Correlations analysis of IFN-γof the mice serum and IFN-γmRNA ofthe mouse auricle in the ACD mice In the ACD group the IFN-γmRNA expression of mouse auricle cells wassignificantly increased. The IFN-γcontent in mice serum was increased too.Xiaofeng powder could reduce not only the levels of IFN-γin the serum butalso decreased the expression of the IFN-γmRNA in the mouse auricle cells.The IFN-γcontent in the serum and the IFN-γmRNA expression in the mouseauricle cells had the consistency. Correlation analysis showed that the IFN-γcontent in the serum was positively correlated with the IFN-γmRNAexpression in the mouse auricle cells. Correlation coefficient was 0.754.Conclusion1. Xiaofeng powder could effectively inhibit allergic contact dermatitis inmice. Itnot only could suppress the partial inflammation response, but alsoinhibit the spleen and thymus increasing reaction with ACD. Xiaofeng powdercould reduce the damage of the mouse skin organization and the cellultrastructure which caused by hypersensitivity.2. Xiaofeng powder could effectively reduce Leukocytes, neutrophilicgranuloeyte and lymphocyte in blood.3. Xiaofeng powder could upregulate the levels of IL-4 in the ACD miceserum and declines its IFN-gamma, the sIL-2R content. Xiaofeng powdercould increase the levels of IL-10 and decrease the levels of IL-13 in thepathological skin organization. Xiaofeng powder also could reduce in thepartial organization the IFN-γmRNA expression; suppress the partial IFN-γprotein translation and the IFN-γsecretion. But the expression of IL-6 mRNAhad no significant effect.4. The levels of IFN-γin the ACD mice serum and IFN-γmRNAexpression in the local tissues of the ACD mice were positively relationship.5. Adjustments to Th1 and Th2 immune balance may be one of themechanisms of inhibiting the ACD on Xiaofeng powder.