| Acute disseminated encephalomyelitis (ADEM) and multiple sclerosis (MS) are common demyelinating diseases in pediatric clinic, the causes of which are unknown, and thought to belong to an autoimmune disease. The process of these diseases are similar to experiment autoimmune encephalomyelitis (EAE), lucubrating to EAE provided us with experiment data ofpathogenesy of them.In the past, the major treating method of ADEM and MS is to utilize nonspecific immunosuppressive agents. Most of these immunosuppressive agents can also inhibit normal function of lymphocyte at the same time of inhibiting autoreactive T lymphocyte. So, it has been research hot spot to search a treating method with low side effect and good treating effect.Oral antigen tolerance is a kind of peripheral tolerance of mature peripheral lymphocyte to oral antigen after organism intake soly protein angigen. Since 1980s, it has become the research hot spot to utilize this character to treat EAE and to research its mechanisms and influencing facts. This study is to investigate the effects of oral antigen tolerance preventing and treating EAE and the changes of IFN-γand IL-10 in rats.Section one: The protective effect of pre-treatment and post-treatment with oral antigen tolerance in rat with EAEObjective: To study the protective effect of oral tolerance on EAE in rats. Methods Sixty-five Wistar rats aged six-eight-weeks were used as research subject and randomly divided into 6 groups:①Group control (CON, n=10);②Group EAE (EAE, n=12);③Group prevention control (PREC, n=8);④Group prevention by oral tolerance (PRE, n=12)⑤Group treatment control (TREC, n=12)⑥Group treatment by oral tolerance(TRE, n=11).Group PRE rats were lavaged with 5mg spinal cord homogenous of guinea-pig on 7,5 and 2 days preimmunizing, the day immunizing and 7, 5 and 2 days after immunizing while the Group PREC rats were lavaged with phosphate balanced solution. Group TRE rats were lavaged with 5mg spinal cord homogenous of guinea-pig on 7 to 13 days after immunizing while Group TREC rats were lavaged with phosphate balanced solution. Group CON rats were not handled at all, rats in other groups were immunized according to the method of EAE model building. All rats were executed after observed for 16 days. Result: 1. Weight increasing: The weight decreased in Group EAE, Group PREC, Group TREC and Group TRE while the body increased obviously and increased slowly in Group PRE. In Group EAE, the body weight decreased (28.77±8.73) g, the increased rate was (-17.23±5.23) %, Group PREC decreased (27.95±12.14)g, the increasing rate was (16.44±7.14)%, Group TREC decreased (29.23±10.42)g, the increasing rate was (-17.4±6.20)% and Group TRE decreased (30.70±16.33) g, the increasing rate was (-18.95±10.08)%, while body weight in Group CON increased (35.21±14.00)g , the increasing rate was (21.34±8.12)%, compared the former four groups with Group CON, there was statistical difference (p＜0.05). But there was no statistical difference among these four groups (p＞0.05) .In Group PRE, body weight increased (7.26±10.75) g, the increasing rate was (4.22±6.25)%, compared with Group PREC, there was statistical difference between them (p＜0.05). 2.Disease incidence: Disease incidence in Group EAE was 83.3%, Group PREC (75%), Group TREC(75%) and Group TRE (81.8%), there was no statistical difference among these four groups (p＞0.05). Disease incidence in Group PRE(33.3%) was lower than Group PREC, there was statistical significance between them (p＜0.05). 3.Average date of accident: Average date of accident in Group EAE was (8.66±0.97)days, Group PREC (7.83±0.41) days, Group TREC (8.44±1.42) days and Group TRE (8.13±0.64) days, there was no statistical difference among these four groups (p＞0.05). Average date of accident in Group PRE was (9.67±1.53)days, compared with Group PREC, there was statistical difference between them(p＜0.05). 4.Maximum neural function score: Maximum neural function score in Group EAE was (2.90±0.74) point, Group PREC (3.00±0.63) point, and Group TREC (2.89±0.60) point differently, there was no statistical difference among these three groups (p＞0.05) .Maximum neural function score in Group PRE was (2.33±0.58)point, compared with Group PREC, there was statistical difference between them (p＜0.05) . Maximum neural function score in Group TRE was (1.75±0.71)point, compared with Group TREC, there was statistical difference between them (p＜0.05) .5.HE staining of pathological section: In Group EAE, Group PREC and Group TREC, myelin substance of rats was infiltrated by generous mononuclear cells, major of which were lymphocyte, infiltrating cells formed type alterations like sleeve; Myelin substance of rats in Group TRE was also infiltrated by a deal of mononuclear cells, major of which were lymphocyte,but no type alteration like sleeve was found; Myelin substance of rats in Group PRE was also infiltrated by few mononuclear cells, major of which were lymphocyte; Myelin substance of rats in Group CON was not infiltrated. Conclusions: There is a preventing and possible a treating effect against EAE in rats with oral tolerance.Section two: The effect of oral antigen tolerance on the concentration of serum IFN-γand IL-10 in rat EAE.Objective: To study the changes of the concentration of serum IFN-γ and IL-10 in rat with EAE and the influence of oral antigen tolerance on it. To approach the role of IL-10 and IFN-γplayed during the process of EAE and oral antigen tolerance. Methods: Sixty-five Wistar rats aged six-eight-weeks were used as research subject and randomly divided into 6 groups. Get the peripheral blood of all rats on the 7,10 and 14 days after immunizing and detect concentration of IL-10 and IFN-γ. Results: On the 7 days after immunizing, average concentration of IL-10 in serum of Group CON was (125.16±25.50) ng/ml,hower, average conentration of IL-10 in Group EAE was (95.19±24.59) ng/ml,Group PREC (85.38±17.42) ng/ml,Group TREC (87.09±19.92) ng/ml,Group TRE (102.60±29.27) ng/ml diffferently, lower compared to Group CON (p＜0.05) and there was no statistical difference among these four later groups (p＞0.05). The concentration of Group PRE was (115.10±18.49) ng/ml, higher than Group PREC (p＜0.05).At the same time, concentration of IFN-γin serum of Group CON was (78.424±8.74) ng/ml, concentration of IFN-γin Group EAE was (122.65±7.86) ng/ml, Group PREC (121.69±5.80) ng/ml, Group TREC (116.29±8.00) ng/ml, Group TRE (116.68±9.73) ng/ml diffferently, higher compared to Group CON (p＜0.05) and there was no statistical difference among these later four groups (p＞0.05) .The concentration of Group PRE was (93.07±5.42) ng/ml, lower than Group PREC (p＜0.05) .On the 10 days after immunizing, concentration of IL-10 in serum of Group CON was (126.26±20.93) ng/ml, concentration of IL-10 in Group EAE was (73.32±24.50) ng/ml, Group PREC (71.38±9.23) ng/ml,Group TREC (71.08±16.45) ng/ml diffferently, lower compared to Group CON (p＜0.05) and there was no statistical difference among these three later groups (p＞0.05) The concentration of IL-10 of Group PRE was (101.69±17.11) ng/ml, higher than Group PREC (p＜0.05). The concentration of IL-10 in Group TRE was (87.06±20.51) ng/ml, higher than in Group TREC (p＜0.05) .At the same time, concentration of IFN-γin Group CON was (77.76±7.10) ng/ml, conentration of IFN-γin Group EAE was (141.48±7.86) ng/ml, Group PREC (138.73±8.63) ng/ml. Group TREC (143.65±10.98) ng/ml diffferently, higher compared to Group CON(p＜0.05)and there was no statistical difference among these later three groups (p＞0.05) .The concentration of Group PRE was (104.39±10.40) ng/ml, lower than Group PREC (p＜0.05). The concentration of Group TRE was (119.79±13.28) ng/ml, lower than Group TREC (p＜0.05) .Fourteen days after immunizing, concentration of IL-10 in Group CON was (124.44±19.64) ng/ml ,and concentration of IL-10 in Group EAE was (83.33±15.72) ng/ml,Group PREC (84.85±9.60) ng/ml,Group TREC (82.17±14.69)ng/ml diffferently, lower compared to Group CON (p＜0.05) and there was no statistical difference among these later three groups (p＞0.05) The concentration of Group PRE was (113.28±20.88) ng/ml, higher than Group PREC (p＜0.05) .The concentration of Group TRE was (106.60±15.80) ng/ml, higher than Group TREC (p＜0.05). At the same time, conentration of IFN-γin Group EAE was (124.03±10.60) ng/ml,Group PREC (121.68±10.07) ng/ml, Group TREC (118.78±11.83) ng/m diffferently, higher compared to Group CON (79.91±9.04) ng/ml (p＜0.05) and there was no statistical difference among these three later groups (p＞0.05) .The concentration of Group PRE (90.83±10.10ng/ml) was lower than Group, PREC (p＜0.05) and Group TRE (103.39±11.36) ng/ml was lower than Group PREC (p＜0.05) . Conelutions: There is disbalance between IFN-γand IL-10 in the serum of rats EAE .This disbalance can be used an index of the early diagnosis to EAE. This disbalance can be retrieved after pre-treatedment or posttreatment with oral antigen tolerance. This may be one of the protecting mechanisms of oral antigen tolerance.
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