| ObjectiveTo generalize syndrome patterns and syndrome factors in unstable angina pectoris coro-nary heart disease patients by statistical and data mining methods and study links between it, the adhesion molecule of P-selectin in serum and concentration variations of VCAM-1. Futherly capture more objective and implicit relations by unsupervised machinery data analy-sis.With the help of ischemia-reperfusion model of human cardiac microvascular endothelial cell to imitate the damage of ischemia of angina pectoris onset on vascular endothelial cell and study the intervention effect of Astragalus Polysaccharide on adhesion molecule such as P-selectin and VCAM-1and its signal transduction mechanism of P38and NF-kb.Method1. Kohonen network clustering was adopted to analyze the symptoms of265unstable angina pectoris (UAP) cases and thus received the common syndrome pattern of UAP. Factor analysis was used to summarize and generalize the syndrome factors. At last, Decision Tree Algorithm was applied for detection of inner connections between syndrome factors, P-selectin and VCAM-1.2. Human cardiac microvascular endothelial cell (HCMEC) was managed by ischemia-reperfusion to imitate the damage of ischemia of angina pectoris onset on vascular endothelial cell. Variations of protein expression of P-selectin and VCAM-1as well as transcription level of mRNA, phosphorylation of P38and NF-Kb in HCMEC in ischemia-reperfusion model intervened by Astragalus Polysaccharide with the help of immunohistochemistry, western botting and quantitative real time PCR assay.Result1. UAP syndrome pattern:According to Kohonen network clustering, the syndrome pattern of UAP were divided into9catigories as below:heart-blood stasis (20.99%), qi deficiency with bood stasis plus phlegm (14.89%), yang deficiency with blood stasis (11.83%), heart-qi deficiency (11.45%), qi and yin deficiency (10.69%), phlegm-turbid obstruction (10.69%), depressed qi transforming into heat (9.16%), phlegm-heat stagnation (6.49%), heart yang deficiency (3.82%)。2. UAP syndrome factor:According to factor analysis, the analysis results of symptoms of UAP patients was received seven common factors (symptom factors) that were blood stasis (47.33%), qi dificiency (38.55%), phlegm turbid (27.10%), heat retention (20.61%) qi stagnation (16.03%), yin deficiency (15.27%) and yang deficiency (14.89%).Forthe57kinds of cyndrome pattern matches; more than ten cases had six factors; five to ten cases had nine factors; three to five cases had eleven factors; two cases had ten factors; one case had twenty-one factors. Among these mathes, bood stasis (15.85%), qi deficiency (12.83%) and qi deficiency with blood stasis (7.17%) were the most frequent patterns.3. The relation between UAP syndrome pattern and adhesion molecules:There were significant differences between P-selectin and VCAM-1concentration of different syndrome patterns (P<0.01). The P-selectin level from lowest to highest in turn:qi deficiency with bood stasis plus phlegm, blood stasis, yang deficiency with blood stasis, phlegm-heat stagnation, depressed qi transforming into heat, phlegm-turbid obstruction, qi and yin deficiency and qi deficiency and yang deficiency. Except for blood stasis group, all other groups had statistical differentces in contrast to qi deficiency with bood stasis plus phlegm group (P<0.05or P<0.01). Besides qi and yin deficiency and yang deficiency group, all other groups showed significant differences in comparison to qi deficiency group. The VCAM-1level from lowest to highest in turn:qi deficiency with bood stasis plus phlegm, phlegm-heat stagnation, blood stasis, depressed qi transforming into heat, yang deficiency with blood stasis, phlegm-turbid obstruction, qi and yin deficiency, qi deficiency and yang deficiency (P<0.05, P<0.01) Except blood stasis, depressed qi transforming into heat and phlegm-heat stagnation group, all other groups had statistical differentces in contrast to qi deficiency with bood stasis plus phlegm group (P<0.05or P<0.01). Besides qi and yin deficiency, phlegm-turbid obstruction and ya deficiency group, all other groups showed significant differences in comparison to qi deficiency group (P＜0.05, P＜0.01)The relation between UAP syndrome factor and adhesion molecules (QUEST algo-rithm):Qi deficiency is related to low P-selectin level rather than VCAM-1level; blood stasis is related to high P-selectin and VCAM-1level; phlegm turbid is linked to high VCAM-1; heat retention is linked to high VCAM-1concentration; yang deficiency is related to low P-selectin level; yin deficiency and qi stagnation had nothing to do with P-selectin and VCAM-1level. Thus, P-selectin had a high expression in blood stasis syndrome while had a low expression in yang deficiency and qi deficiency syndrome; VCAM-1expressed highly in blood stasis, heat retention and phlegm turbid syndrome pattern while expressed low in yang deficiency syndrome.4. Protein expression and mRNA transcription level of adhesion molecules and P38, NF-Kb by ischemia-reperfusion:It was observed that protein expression as well as mRNA transcription level of P-selectin and VCAM-1were higher than normal group (P＜0.05, P<0.01). While the mRNA transcription level of P-P38and its target ATF-2as well as NF-Kb and its target ixb were significantly elevated in contrast to normal group (P<0.01)5. Protein expression and mRNA transcription level of adhesion molecules and P38, NF-Kb after intervention by Astragalus Polysaccharide:Protein expression as well as mRNA transcription level of P-selectin and VCAM-1of model group were obviously down-regulated than control group (P＜0.05, P<0.01). While the mRNA transcription level of P-P38, ATF-2and kb mRNA dropped drastically than model group (P<0.01). However, in comparison to PDTC, the inhibitor of NF-Kb and to SB203580which is the inhibitor of P38, there was no statistical difference (P>0.05)Conclusion1. In UAP patients, heart-blood obstruction and qi deficiency with bood stasis plus phlegm sydrome were the most common ones.2. The syndrome factors of UAP were blood stasis, qi dificiency, phlegm turbid, heat retention, qi stagnation, yin deficiency and yang deficiency and they were commonly observed in combinated forms. The rule of combination was complicated and blood stasis, qi deficiency and qi deficiency with blood stasis were the most popular ones.3. P-selectin and VCAM-1had a high expression in intermingled deficiency and exces-sive syndrome and excessive syndromes, especially in qi deficiency with bood stasis plus phlegm syndrome. In additon, they had a low expression in deficiency syndromes and their expression were lowest in yang and qi deficiency synreome pattern. Thus it was speculated that the concentration of P-selectin reflected the degree of blood stasis directly while reflected the degree of phlegm-turbid indirectly. As for VCAM-1concentration, it mirrored phlegm-turbid directly while mirrored blood stasis and heat retention level indirectly. Heat retention promoted the density of P-selectin and VCAM-1and its effect was greater than the force of qi stagnation.4. In HCMEC under normal cultivation circumstance, the activity of P38and NF-Kb were inhibited and P-selectin and VCAM-1were in low expression and transcription level. After treated by ischemia-reperfusion, activity of P38and NF-Kb in HCMEC enhanced sub-stantially. In the mean time, expression and transcription level of P-selectin and VCAM-1promoted significant, causing further damage cellular injury.5. After intervened by Astragalus Polysaccharide, protein expression as well as mRNA transcription level of P-selectin and VCAM-1in HCMEC down-regulated obviously which saved as inhibtor of P38and NF-Kb with similar effects as PDTC and SB203580. Thus, it was speculated that Astragalus Polysaccharide was of impressive effects on signaling pathway of P38together with signal transduction of NF-Kb. Additionally, the concentration-dependent effect was achieved by declined the activity of pathways mentioned above leading to dropped level of related adhesion molecules as P-selectin and VCAM-1. Finally, the adhesion mole-cule mediated endothelial cell and white cell adhesion was therefore blocked and thus white cells were arrested from releasing cytotoxic mediators to damage the endothelial cells.In the study of UAP patients, blood stasis and qi dificiency were the most common ones. P-selectin and VCAM-1had a high expression in intermingled deficiency and excessive syn-drome and excessive syndromes. In additon, they had a low expression in deficiency syndromes. After hypoxia reperfusion injury, the high expression of P-selectin, VCAM-1concentrations decreased significantly after intervention by astragalus polysaccharide, presumably astragalus have both supplementing qi and activating blood circulation, and plays a very important role in the treatment of UAP.
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