| Subject:With the technology of Proteomics analysis, we extracted and separated total protein of Peripheral blood of the healthy volunteers and chronic hepatitis B (CHB) in TCM with Internal Stasis of Dampness and Heat Syndromes (Ⅱ), Depressed Liver With the Insufficient Spleen Syndromes (Ⅲ), Collateral Obstruction by Blood Stasis Syndromes (Ⅵ), liver and kidney yin deficiency syndrome (Ⅴ) and the spleen and kidney yang deficiency (Ⅳ) five syndromes, initially explored the various TCM syndromes of chronic hepatitis B between inflammatory reaction conditions laws proteomics, clinical recognition of TCM Syndrome Chronic inflammatory conditions provide useful indicators of objective and, ultimately, to further explore the essence of TCM syndrome of chronic hepatitis B to provide a scientific basis. Methods: According to the "Hepatitis TCM standards", in Luzhou Medical College Hospital of Chinese Medicine in October 2009 -2010 12-month period in patients with chronic hepatitis B in the collection of Internal Stasis of Dampness and Heat Syndromes, Depressed Liver With the Insufficient Spleen Syndromes, Collateral Obstruction by Blood Stasis Syndromes, liver and kidney yin deficiency syndrome and the spleen and kidney yang deficiency five syndromes five syndromes as the observation group were 80 cases to healthy controls 5 cases of the control group. Using two-dimensional electrophoresis technology in PH4-7 in the dry IPG gel strips to about 130μg of sample volume,75000v.h electrophoresis conditions (the initial establishment of chronic hepatitis B in the two-dimensional electrophoresis system for plasma. And get a good resolution differences in protein expression patterns.) scan gel imaging analysis software to more than ImageMaster2D plasma protein expression profiles. Standardization of the experimental group and control group, the total gray value (% Vol) a difference of up to 2 times more protein spots differentially expressed protein as a point, cut the corresponding differentially expressed proteins, the matrix-assisted laser desorption time of flight mass spectrometry (MADLDI-TOF -MS) detection, combined with 2-DE map of the relative points of each protein isoelectric point (PI), relative molecular weight (Mw) and the Swiss-prot protein database search match and identified the differentially expressed proteins. Hepatitis B screening in five syndromes were apparent regularity in the differential expression of liver inflammation and can reflect the status of the protein, in expanding the sample size syndrome by ELISA under the above validation of differentially expressed proteins. Measurement data with the (x±S) said, with SNK-q statistical analysis, all data in the computer by SPSS v15.0 statistical software. Results:(1) to obtain good resolution and reproducibility of plasma proteome than two-dimensional electrophoresis silver staining. (2) by comparing healthy control group, Internal Stasis of Dampness and Heat Syndromes, Depressed Liver With the Insufficient Spleen Syndromes, Collateral Obstruction by Blood Stasis Syndromes, liver and kidney yin deficiency syndrome and the spleen and kidney yang deficiency two-dimensional electrophoresis map of the five syndromes were apparent regularity in the differentially expressed protein spots were five, were identified by mass spectrometry Haptoglobin (HP), immunoglobulin j chain (Ig-j), retinol binding protein (RBP), C-reactive protein (CRP), glass with protein (VN). Its main function involves decreased hepatic synthetic function, inflammation, immune pathologic injury. (3) hepatitis B screening in five syndromes were apparent regularity in the differential expression of liver inflammation and can reflect the status of the protein CRP, RBP, and VN, in expanding the sample size syndrome by ELISA of the above cases, differential expression of proteins to verify that retinol binding protein (RBP) has fallen relative to the normal group, the lower order (maximum reduction, the top)Ⅵ>Ⅴ>Ⅳ>Ⅲ>Ⅱ. Each group and the normal group were significantly different (P<0.05), comparison between groups IV and III there were significant differences (P<0.05); glass with protein (VN) relative to the normal group were dropped, reducing the order of (reduce the maximum, the top)Ⅵ>Ⅴ>Ⅳ>Ⅱ>Ⅲ. Each group and the normal group were significantly different (P<0.05), comparison between groups VI and V had significant differences (P<0.05),ⅡandⅢare significantly different (P<0.05); C-reactive protein (CRP) relative to the normal group are higher (higher) order (higher up, the top)Ⅵ>Ⅴ>Ⅳ>Ⅲ>Ⅱ. Each group compared with the normal group were significantly different (P<0.05), VI and V between the two groups was significant difference between (P<0.05). Conclusions:(1) the initial establishment of chronic hepatitis B with Internal Stasis of Dampness and Heat Syndromes, Depressed Liver With the Insufficient Spleen Syndromes, Collateral Obstruction by Blood Stasis Syndromes, liver and kidney yin deficiency syndrome and the spleen and kidney yang deficiency and healthy control group, the plasma proteome expression profiles. (2) TCM five chronic hepatitis B and normal expression of multiple different proteins, indicating that there are multiple abnormal expression of CHB, this is the traditional Chinese medicine "card " the essence of the scientific basis (3) Expand the sample size syndrome in the case of the above by ELISA, differentially expressed proteins CRP, RBP, and VN verify that retinol binding protein (RBP) has fallen relative to the normal group, the lower order (maximum reduction, the top)Ⅵ>Ⅴ>Ⅳ>Ⅲ>Ⅱ. Each group and the normal group were significantly different (P<0.05), comparison between groupsⅣandⅢthere were significant differences (P <0.05); glass with protein (VN) relative to the normal group were dropped, reducing the order of (Reduce the maximum, the top)Ⅵ>Ⅴ>Ⅳ>Ⅱ>Ⅲ. Each group and the normal group were significantly different (P<0.05), comparison between groupsⅥand V had significant differences (P<0.05),ⅡandⅢare significantly different (P<0.05); C-reactive Protein (CRP) relative to the normal group are higher (higher) order (higher up, the top)Ⅵ>Ⅴ>Ⅳ>Ⅲ>Ⅱ. Each group compared with the normal group were significantly different (P <0.05),ⅥandⅤbetween the two groups was significant difference between (P <0.05). This suggests that CRP, RBP, and VN changes of TCM syndrome and chronic hepatitis B have some relevance. VN expression was reduced and reduced and reduced RBP expression consistent with the order to reduce the amount of Collateral Obstruction by Blood Stasis Syndromes highest card, followed by liver and kidney yin deficiency syndromes, the spleen and kidney yang deficiency syndromes, Depressed Liver With the Insufficient Spleen Syndrome, Internal Stasis of Dampness and Heat Syndromes reduce the lowest, suggesting that in the CHB with Collateral Obstruction by Blood Stasis Syndromes in the impaired liver function the most important, followed by liver and kidney yin deficiency syndromes, the spleen and kidney yang deficiency syndromes, Depressed Liver With the Insufficient Spleen Syndrome, Internal Stasis of Dampness and Heat Syndromes Relatively mild impairment(4) CRP increased relative to the normal group order (higher up, the top)Ⅵ>Ⅴ>Ⅳ>Ⅲ>Ⅱand RBP decrease relative to normal sequence (lower row up front)Ⅵ>Ⅴ>Ⅳ>Ⅲ>Ⅱand VN reduced the relative order of the normal group (maximum reduction, the top)Ⅵ>Ⅴ>Ⅳ>Ⅱ>Ⅲconsistency, suggesting that in chronic hepatitis B in five Syndrome, liver damage liver inflammation and its positive correlation.
|
PDF Full Text Request