The Study Of Nuclear Proteomics For Lupus Mice Affected By The Recipe Of Nourishing Yin And Reducing Heat

Background and Objectives:Systemic lupus erythematosus is a frequent autoimmune disease and its etiology remains obscure.Many studies discovered that polygene abnormity of SLE is displayed the level of multipotent hemopoietic stem cell,then which turn into abnormal lymphocyte,so it is proposed that SLE maybe belongs to stem cell disease of multiclone dysfunction,now radical cure is impossible,the most patients require to take glucocorticoid for lifetime,but the adverse reactions of long-term medicine greatly will effect life quality and survival rate.in our country clinical practice prove conbined treatment of traditional Chinese medicine and western medicine decrease obviously side reaction of Western medicine.Now scholars have studied the therapeutic effect and mechanism in many aspects such as cytokine,apoptosis,immunocyte and hypothalamus-pituitarium -endocrine axis.so it is a present tendency of investigation to explain mechanism of the traditional Chinese medicine from molecular level by using advanced molecular biology technique combining to theory of traditional Chinese medicine.Proteome technique is not only identify the autoantibody mode of autoallergic disease,but also investigate autoimmune diversity;In addition,It can assess efficiently protein target of autoimmune response.enhanced laser desorpion / lonizaion time-of-fight mass spectrometry(SELDI—TOF-MS)is advanced technology.Our division have studied traditional Chinese medicine to treating SLE in the past many years,wu think the basic pattem of syndrome of SLE is the asthenic kidney-YIN and yin asthenia generating intrinsic heat,So eliminating heat by nourishing kidney-YIN is the rule of treating SLE.The prophase research have demonstrated the recipe of nourishing Yin and reducing Heat can decrease dosage of glucocorticoid and relieve adverse reaction of glucocorticoid.The topic of nationnatural science fund indicated the recipe of nourishing Yin and reducing Heat for patients may adjust the genetic expression and proteinaceous expression to achieve therapeutic action.Our objectives are to measure bone marrow cell and lymphocytic nucleoprotein array of lupus mice of different groups by proteome technique and then compare the distinction of each group mass spectrographic.At last wu try to reveal the mechanism of the recipe of nourishing Yin and reducing Heat and associating prednisone.Methods and Materials:1.experimental animal:female MRL/LPR lupus mice,body weight 25.5—35.5 g,average 30.6 g2.drug composition:FRUCTUS CORNI15 rehmannia dride rhizome20 indian bread15 orientale Juzep 15 CORTEX MOUTAN15 abrotani herba8 Glycyrrhiza uralensis63.Methods:control group:isotonic Na chloride 0.6ml,intragastric administration once per day.prednisone group:prednisone 9mg/kg/d(equivalently 1mg/kg/d for human),to match suspl.0.6ml by isotonic Na chloride intragastric administration once per day.the recipe of nourishing Yin and reducing Heat group:dosage of the traditional Chinese medicine is 0.02mi/g/d,together 0.6ml apozem.Prednisone and the recipe of nourishing Yin and reducing Heat group:prednisone is 7mg/kg/d(equivalently 0.8mg/kg/d for human),to match suspl.0.2ml by isotonic Na chloride,intragastric administration once per day;:dosage of the traditional Chinese medicine is 0.2ml apozem.lupus mice were fed by the water and the feedstuff of athymic mouse up to 12 weeks age and then used different medicine by tragastric administration for 5weeks.died mice was rejected。4.to take samples:blood serum,lymphocytes of the spleen,bone marrow cell5.to extract nucleoprotein6.to take nucleoprotein express spectra of bone marrow cell and lymphocytes of the spleen by SELDI method7.data processing and Statistical analysis8.Proteinaceous contradistinction and analytical methodResults:The result of bone marrow cell nucleoprotein:1)To compare control group with Western medicine group we found there are 15 differential expression protein peak of statistical significance(P＜0.05),of the total control group protein peak having 11 pieces,Western medicine group protein peak having 4 pieces;to compare control group with Chinese Western medicine group we found there are 14 differential expression protein peak of statistical significance(P＜0.05),of the total control group protein peak having 10 pieces,Chinese Western medicine group protein peak having 4 pieces.They were both similar or identical protein peak of 10 pieces. A.Comparing Western medicine group,Protein peaks of 2555,2756,3048,3090,3406,4722,5076,10147,2907,5503,6480 were statistical significance in Control group. Comparing Chinese Western medicine group,Protein peaks of 2556,2757,3050,3093,3409,4722,5078,10152,4479,6510 were statistical significance in Control group.They were both similar or identical protein peaks of 8 pieces.Different protein peaks were2907,5503,6480和4479,6510.It is focal point to search possible proteinaceous differences corresponding protein peaks.B.Comparing Control group,Protein peaks of 3063,10252,10476,13092 were statistical significance in Western medicine group.Comparing Control group,Protein peaks of 11159,12228,10480,13098 were statistical significance in Chinese Western medicine group.They were both similar or identical protein peaks of 2 pieces.Different protein peaks were 3063,10252和11159,12228.It is focal point to search possible proteinaceous differences corresponding protein peaks.2)To compare Chinese Western medicine group with Western medicine group we found there are 1 protein peak of statistical significance(P＜0.05)belonging to Western medicine group,The others were all P＞0.1.3)To compare Traditional Chinese medicine group with Western medicine group we found there are 5 protein peak of statistical significance(P＜0.065),The others were all P＞0.1.Protein peak of 3245 belonged to Traditional Chinese medicine group;Protein peaks of 13073,13235,13276,13390 belonged to Western medicine group.4)To compare Traditional Chinese medicine group with Chinese Western medicine group we found there are 1 protein peak of statistical significance(P＜0.065)belonging to Chinese Western medicine group,The others were all P＞0.1.5)To compare Control group with Traditional Chinese medicine group we found there are 3 protein peak(2557,3247,5078)of statistical significance(P＜0.05).The result of splenic Iymphocyte nucleoprotein:1)To compare control group with Western medicine group we found there were 17 protein peak of statistical significance(P＜0.05),of the total protein peaks of control group having 12 pieces,Western medicine group protein peaks having 5 pieces.To compare control group with Chinese Western medicine group we found there were 12 protein peak of statistical significance(P＜0.05),of the total protein peak of control group having 5 pieces,Chinese Western medicine group protein peak having 7 pieces. A.Comparing Western medicine group,Protein peaks of 2896,2953,3080,3100,2917, 2784,2967,3053,3158,3842,8458,8658 were statistical significance in Control group.Comparing Chinese Western medicine group,Protein peaks of 2899,2956,3083,3101,4237 were statistical significance in Control group.They were both similar protein peaks of 4 pieces.Different protein peaks were 2917,2784,2967,3053,3158,3842,8458,8658and4237.It is focal point to search possible proteinaceous differences corresponding protein peaks.B.Comparing Control group,Protein peaks of 3699,5327,5405,12335,12418 were statistical significance in Western medicine group.Comparing Control group,Protein peaks of 6526,6551,6580,6605,6705,6724,12235 were statistical significance in Chinese Western medicine groupThey were not similar protein peaks.We searched possible proteinaceous differences corresponding protein peaks.2)To compare Chinese Western medicine group with Western medicine group we found there were 2 protein peak of statistical significance(P＜0.066)protein peak 4150,6186 belonging to Western medicine group,The others were all P＞0.1.3)To compare Traditional Chinese medicine group with Western medicine group we found there are 7 protein peak of statistical significance(P＜0.065),Protein peak of 3053, 4285,8610 belonging to Traditional Chinese medicine group and Protein peaks of 5403, 4106,6028,9977 belonging to Western medicine group.The others were all P＞0.1.4)To compare Traditional Chinese medicine group with Chinese Western medicine group we found there are 4 protein peak of statistical significance(P＜0.07),protein peak of 6579,6629 belonging to Chinese Western medicine group and protein peaks of 4232, 4289 belonging to Traditional Chinese medicine group.5)To compare Control group with Traditional Chinese medicine group we found there were 7 protein peak of statistical significance(P＜0.06).Comparing Traditional Chinese medicine group,Protein peaks of 2951,3889,3902,8486 were statistical significance in Control group;Comparing Control group,Protein peaks of 5691,6573,8560 were statistical significance in Traditional Chinese medicine group.We searched possible proteinaceous differences corresponding protein peaks.Conelusions:1)The nucleoprotein of bone marrow cell and splenic lymphocyte shew many significant Protein peaks after applying Prednisone and the traditional Chinese medicine to lupus mice.It indicated Prednisone and/or the traditional Chinese medicine maybe interfere myeloid multipotent hemopoietic stem cell differentiation in bone marrow and effect protein express so as to achieve therapic purpose.It is the same as experimental hypothesis. 2)To compare control group with Western medicine group,we found there were some proteins in Control group which aggravate course and increase cytokine secretion,there were some proteins in Western medicine group which adjust immunological activity and rivalry apoptosis.It indicated the mechanism of prednisone suppress the passage of nuclear factor-NF,strengthen the function of tumor growth factor(TGF-β),diminish chemotatic factor,maintain the homeostasis of the hypothalamus-pituitarium-endocrine axis,suppress Apoptosis and reinforce mitochondrial function.3)To compare Chinese Western medicine group with Western medicine group we found there are 1 protein peak of statistical significance in the nucleoprotein of bone marrow cell;To compare Chinese Western medicine group with Western medicine group we found there are 2 protein peaks of statistical significance in the nucleoprotein of splenic lymphocytes.It explained that Chinese Western medicine and Western medicine effecting nucleoprotein of bone marrow cell and splenic lymphocytes was semblable. The prednisone dosage of Chinese Western medicine group was 80%of the dosage of Western medicine group,so wu predict the recipe of nourishing Yin and reducing Heat and prednisone have synergistic action.4)To compare control group with traditional Chinese medicine group,we found there are 3 protein peak of statistical significance in the nucleoprotein of bone marrow cell.The protein of traditional Chinese medicine group have nothing to do with immunization.It demonstrated traditional Chinese medicine merely have bad therapeutic efficacy.5)Wu can not discover the synergistical pathway of prednisoneand the recipe of nourishing Yin and reducing Heat in the experiment6)Wu discovered many protein peaks unresponding protein in the experiment,wu need study deeply for future.