The Interrelated Sdudy Between Tcm Syndromes And Clinical Pathology In Lupus Nephritis And Proteomic Research Of Lupus Nephritis

Objectives1. Clinical StudyThe study established observation tables and databases of lupus nephritis with TCM syndrome elements,physicochemical index and pathological types etc through the collection of lupus nephritis patients in general,TCM syndromes, physicochemical index and renal pathology, then analysed the relationship of TCM syndromes and physicochemical index,pathological types of lupus nephritis by using statistical analysis software, to provide evidence for microcosmic differentiation of TCM syndromes,which was beneficial to guide us for the diagnosis and treatment of LN with Integrated Traditional Chinese and Western Medicine.2. Experimental StudyThe study analysed serum proteomics of normal human and patients with lupus nephritis and lupus nephritis patients with excessive heat-toxic and yang deficiency of spleen and kidney, screening differential expression proteins, and try to found proteins that associated with lupus nephritis and important application perspective biomarkers, as well as to found proteins that related with TCM syndromes,then to explore the protein expression profiles and relationship between proteins and TCM types. To try to provide objective basis for syndrome differentiation and treatment of lupus nephritis.Methods1. Clinical studies120cases of patients with lupus nephritis were divided into eight different TCM syndromes including (excessive heat-toxic,yin deficiency of liver and kidney,Qi deficiency of spleen and kidney,yang deficiency of spleen and kidney,deficiency of both Qi and Yin,wind-dampnes internal disturbance, blood stasis and Damp-heat), and clinical physicochemical index and renal pathology collection, and then used SPSS statistical analysis software to analyse the relationship between TCM types and physicochemical index,renal pathological types of patients with lupus nephritis.2. Experimental StudyTo separate serum proteins by using two-dimensional gel electrophoresis for protein separation on lupus nephritis patients and normal persons and lupus nephritis with excessive heat-toxic and Yang deficiency of spleen and kidney, the gels were dyed with silver staining and scanned for gel pictures, to find out the differential proteins with software. The differential proteins were cut and digested with trypsin and identificated them with mass spectrographic analysis, and to obtain the finger print map of peptide, then to serch them through Protein Data Bank and identificate them.Results1. Clinical studiesThe study selected120cases, according to the general condition of patients, symptoms, signs, physicochemical index and renal pathology were compared and analysed the relationship of TCM types and physicochemical index, renal pathology, as follows:(1) General data analysis:120cases of patients with lupus nephritis,14cases were male, aged20years to38years, mean age was27.6±5.2years old, average disease course was28.3±6.5months;106cases were female, aged16years to58years, mean age was32.5±8.6years, average disease course was36.7±9.3months.(2) The distribution of TCM types:The frequency analysis showed that Qi deficiency of spleen and kidney was the major type, there was35cases (29.2%); yin deficiency of liver and kidney,excessive heat-toxic,deficiency of both Qi and Yin,yang deficiency of spleen and kidney, respectively there was26cases (21.7%),16cases (13.3%),14cases (11.7%),13cases (10.8%), wind-dampness internal distrubance was16cases (13.3%); blood stasis and damp-heat types, respectively was56cases (46.7%) and41cases (34.2%).(3) Pathology types:The frequency analysis showed that type Ⅳ was the major type, there was87cases (72.5%), followed by the type Ⅴ+Ⅳ, type Ⅴ and type Ⅲ, there were10cases (8.3%),8cases (6.7%),8cases (6.7%); the type Ⅴ+Ⅲ was6cases (5%), type Ⅱ was5cases (4.2%), type Ⅰ and type Ⅵ each had4cases (3.3%) and2cases (1.7%). Using Spearman rank correlation analysis, results suggested that TCM types and renal pathology types without correlation.(4)Clinical types:The frequency analysis showed that nephrotic syndrome was the major type,72cases (60%), chronic nephritis there was36cases (30%), latent nephritis and rapidly progressive glomerulonephritis respectively was6cases (5%) and4cases (3.3%). Using Spearman rank correlation analysis, results suggested that TCM types and clinical types without correlation.(5) Immune index:The frequency analysis results showed that120cases of lupus nephritis patients with ANA positive (100%); Anti ds-DNA positive was101cases (84.2%), anti-Sm positive was41cases(34.2%), anti-ssA positive was55cases (45.8%), the anti-ssB positive was33cases(27.5%), serum C3decreased was104cases (86.7%), the ANCA positive only had4cases(3.3%). Using Spearman rank correlation analysis, results suggested no correlation between TCM types and immune index.(6) Lupus disease activity level:The frequency analysis results showed that the lowest score of SLE-DAI integral was4points, the highest was28points, the average score was (16.28±5.46) points. Severe activity lupus (SLE-DAI>15)there was70cases (58.3%), followed by moderate activity was33cases (27.5%), mild activity was14cases (11.7%), no activity only had3cases (2.5%). Using Spearman rank correlation analysis, results suggested no correlation between TCM types and disease activity types.Using single factor statistical analysis of variance on TCM types and SLE-DAI integrals, the results suggested:the SLE-DAI integral of deficiency of both Qi and Yin type was the lowest (12.69±5.06), significantly less than excessive heat-toxic and Yang deficiency of spleen and kidney (P<0.05). the SLE-DAI integral of wind-dampness internal distrubance less than Yang deficiency of spleen and kidney (P<0.05). The SLE-DAI integral between other TCM types had no statistical significance (P>0.05).(7) Clinical manifestations:The frequency analysis results showed that the most common damage of the extra-renal organs was the blood system, there was92cases (76.7%), fever in36cases (30%),49cases of arthritis (40.8%), facial erythema was44cases (36.7%), oral ulcers was25cases (20.8%), light-sensitive was25cases (20.8%), serositis was18cases (15%), nervous system damage was6cases (5.0%). Using Spearmanrank correlation analysis, results suggested that TCM types and clinical manifestations without correlation.(8)Physicochemical indicators:The results showed that physicochemical indexes of WBC, TG, LDL-C and HDL-C in groups of differential TCM types have no difference;the LYM of Excessive heat-toxic type was significantly lower than Yin deficiency of liver and kidney and Qi deficiency of spleen and kidney, LYM of yang deficiency of spleen and kidney group was significantly lower than Yin deficiency of liver and kidney, other groups showed no differences;the RBC of yang deficiency of spleen and kidney group was significantly lower than Yin deficiency of liver and kidney and deficiency of both Qi and Yin, other groups showed no significant difference; the HGB of yang deficiency of spleen and kidney group was significantly lower than Yin deficiency of liver and kidney, other groups showed no significant difference;the PLT of deficiency of both Qi and Yin group was significantly higher than damp-heat group, other groups had no significant difference; the FIB level of blood stasis type was significantly higher than the other groups;the ALB level of Yang deficiency of spleen and kidney group significantly lower than the types of yin deficiency of liver and kidney, Qi deficiency of spleen and kidney, wind-dampness internal disturbance and damp-heat,and the ALB level of blood stasis group significantly lower than yin deficiency of liver and kidney, the other groups had no obvious difference; the levels of BUN, CREA, UA and Cys C of Yang deficiency of spleen and kidney were significantly higher than those of other groups, the other groups there were not significantly different; the quantitates of24hour urine protein of Yin deficiency of liver and kidney group was significantly lower than that yang deficiency of spleen and kidney and blood stasis groups, other groups had no significant difference. These results may suggest that TCM types have relavent with LYM,RBC,HGB,PLT,FIB,BUN,CREA,UA,Cys C and the quanti tates of24hour urine protein.2.Experimental StudyIn the study we selected4cases of the normal persons,4cases of excessive heat-toxin type of LN patients and4cases of Yang deficiency of spleen and kidney type of LN patients, by using two-dimensional gel electrophoresis for protein separation. The experimental results showed:the excessive heat-toxin type of LN patients had5differential expression spots contrast to Yang deficiency of spleen and kidney of LN patients, the excessive heat-toxin type of LN patients high expressed of the proteins were Ig mu chain C and Inter-alpha-trypsin inhibitor heavy chain H4, yang deficiency of spleen and kidney type of LN patients high expressed of the protein was Keratin type1cytoskeletal10. the LN patients had4differential expression spots contrast to normal persons, LN patients high expressed of the proteins were Alpha-1-antitrypsin and Alpha-1-antichymotrypsin, normal persons high expressed of the protein was Protein disulfide-isomerase A6.Conclusion1. Clinical research(1) The TCM types have no correlation with renal pathology, clinical classification, immune indicators and clinical manifestations in lupus nephritis patients.(2)The SLE-DAI is correlated with the TCM types in Lupus Nephritis.(3)The TCM types have relevant to physichemical indicators such as blood LYM,RBC,HGB,PLT,FIB,BUN,CREA,UA,Cys C and urinary protein; and have no significant correlation with blood WBC,TG,LDL-C and HDL-C.2. Experimental StudyThere are some difference of proteins expressing between normal persons and Lupus Nephritis patients, the different proteins may help to reveal the pathogenesis of lupus nephritis or as new biomarkers for lupus nephritis, it is important in the diagnosis and treatment of lupus nephritis;and also there are some difference of proteins expressing between excessive heat-toxic and yang deficiency of spleen and kidney in Lupus Nephritis patients, we may can use proteomics technology to reveal the mechanism of TCM.