Effects Of Tongxinluo On VCAM-1 And PAI-1 Expression In Atherosclerotic Rabbits

Background and PurposeAtherosclerosis has become a threat to human health, which is one of the major diseases causing the death. It can lead to the damage of more than one target organ, seriously affecting people's quality of life. To solve the problems fundamentally caused by atherosclerosis, a lot of basic experiments and clinical observation are committed to the pathogenesis of atherosclerosis and treatment research. By buiding atherosclerosis model of rabbits with high fat diet, this study aims at understanding expression of the PAI-1,the VCAM-1 in rabbit serum and lesions in the local atherosclerosis, observing effect of TXL on PAI-1 and VCAM-1 and its anti-atherosclerosis, mechanism of action.Materials and Methods1. Building up atherosclerosis models and examining the expression of VCAM-1 and PAI-1 in plasma and arterial walls in the normal diet and the cholesterol diet groups by means of ELISA and immunohistochemistry analysis.1.1 Atherosclerosis models'Building-up1.1.1 Animals and groupsTwenty four male rabbits were randomly divided into the normal diet, the cholesterol diet and the TXL groups. Eight rabbits for each group and fed for 14th week.1.1.2 Making pathology specimensAfter 14th week, the rabbits were put to death and the aortas were harvested for the pathologic morphology observations.1.1.3 Establishing hypercholesterolemiaThere was no obvious difference among TC,TGand LDL in the normal diet and the cholesterol diet groups in plasma in 0 week(P>0.01)and there was no obvious difference between TC,TG and LDL in the normal diet group in 14th week compared with 0 week(P>0.01).There was an obvious increase of TC,TG and LDL in the cholestrol diet group after 14th week and there were obvious differences compared with 0 week(P<0.01).1.2 Measuring the expression of VCAM-1 and PAI-1 in the normal diet and the cholesterol diet groups in plasma by mean of ELISA1.2.1 Expression of VCAM-1 There was no significant difference in VCAM-1 level in the normal diet group in 0 and 14th week(P>0.01).VCAM-1 level increased in the cholesterol diet group in 14th week compared with 0 week(P<0.01).1.2.2 Expression of PAI-1There was no significant difference in PAI-1 level in the normal diet group in 0 and 14th week (P>0.01).PAI-1 level increased in the cholesterol diet group in 14th week compared with 0 week(P<0.01).1.3 Measuring the aorta expression of VCAM-1 and PAI-1 in the normal diet and the cholesterol diet groups by mean of immunohistochemistry analysis1.3.1 Expression of VCAM-1A little brown positive stained substances were found in the intimas and the mediums in the normal diet group and the positive percentage was 2.92±0.31%. A great number of brown positive stained substances were found in the intimas and the mediums in the cholesterol diet group and the positive percentage was 18.38±2.55%.The positive percentage of the substances in the cholesterol diet group obviously increased compared with the normal diet group(P<0.01).1.3.2 Expression of PAI-1A little brown positive stained substances were found in the intimas and the mediums in the normal diet group and the positive percentage was 3.24±0.21%.Lots of brown positive stained substances were found in the intimas and the mediums in the cholesterol diet group and the positive percentage was 15.46±2.23%.The positive percentage in the cholesterol diet group obviously increased compared with the normal diet group(P<0.01).2.Regulating lipid effect of TXLThere was no obvious difference of TC,TG and LDL in three groups in plasma in 0 week(P>0.05),and there was no obvious difference of TC,TG and LDL in the normal diet group in 14th week compared with 0 week(P>0.05).TC,TG and LDL of the cholesterol diet and the TXL groups in 14th week were obviously higher than normal diet group (P<0.01).TC,TG and LDL of the TXL group were lower than the cholesterol diet group in 14th week (P<0.01).3 Effects of TXL on VCAM-1 and PAI-13.1 Effect on plasma3.1.1 Effect on VCAM-1There was no obvious difference in the three groups in 0 week(P>0.01).VCAM-1 in the cholesterol diet and the TXL groups in 14th week were higher than in the normal diet group in 0 week(P<0.01).VCAM-1 of the TXL group was lower than the cholesterol diet group in 14th week(P<0.01).3.1.2 Effect on PAI-1There was no obvious difference in the three groups in 0 week(P>0.01);PAI-1 in the cholesterol diet and the TXL group in 14th week were higher than the normal diet group in 0 week(P<0.01).PAI-1 of the TXL group was lower than the cholesterol diet group in 14th week (P<0.01).3.2 Effect of TXL in the positive percentage of VCAM-1 and PAI-1 in the aorta walls3.2.1 Effect on VCAM-1Lots of brown positive stained substances were found in the intimas and the mediums in the cholesterol diet and the TXL groups and they were 18.38±2.55%and 13.18±1.25%.The quantities of the substances in the TXL group were lower than the cholesterol diet group(P<0.01).3.2.2 Effect on PAI-1Lots of brown positive stained substances were found in the intimas and the mediums in the cholesterol diet and the TXL groups and they were 15.46±2.23%and 9.45±1.09%. The percentage of the substances in the TXL group was lower than that of the cholesterol diet group(P<0.01).These results are expressed by mean±standard deviation (±S) that own and control groups using t test, P<0.05 significant difference.Results1.There was an obvious increase of TC,TG and LDL in the cholestrol diet group and the TXL group after 14th week (P<0.01), and there exised obvious differences compared with 0 week(P<0.01).TC,TG and LDL in the TXL group are less than the cholestrol diet group(P<0.01).2. The PAI-1 and the VCAM-1 levels in the cholesterol diet group and TXL group after 14th week were significantly higher compared with 0 week (P<0.01), and there exised obvious differences compared with 0 week(P<0.01). The PAI-1 and the VCAM-1 levels in the TXL group were significantly lower than cholesterol diet group (P<0.01).3. Immunohistochemical analysis showed that we can and seen a lot of brown-yellow membrane staining granular substance had been seen in the intimal thickening in cholesterol diet group, partial integration into the film, especially in endometrial. However staining substance in the TXL group was significantly less than the cholesterol diet group, three groups were significantly different (P<0.01).Conclusion1.The expression of VCAM-1 and PAI-1 in serum increase in the rabbits of AS;the percentage of the positive stained of VCAM-1 and PAI-1 is much higher in the rabbits of AS. VCAM-1 and PAI-1 over-expression may be one of mechanisms of AS.2.TXL reduces the percentage of expression of VCAM-1 and PAI-1 in serum in the rabbits of AS,and inhibits the percentage of positive stained of VCAM-1 and PAI-1 may be one mechanism of AS.3.TXL has greatly effect of regulating TC,TG and LDL,as well as on mitigation and suppression of AS.4. Measuring VCAM-1 and PAI-1 in serum has great significance for early diagnosis of AS disease and monitoring of AS'occurrence, development and prognosis.