Based On The As Role Of The Inflammatory Response In Sl Extract Control And Its Influence On Cd18 Expression Study

Atherosclerosis(AS), the major cause of ischemic cerebrovascular disorder(CVD) destroying human health,is a multifactorial pathological process aorta and medium -sized artery.The lipid metabolism disorder like hyperlipemia, the blood vessel endothelium impairment,the inflammatory reaction were believed as the basis, initiation and development of AS respectively. Although AS mechnisms were compli -cated, the damage-reaction theory and inflammation theory proposed by professor Ross were popular because those theories were complemented with lipid infiltrating theory, explainning the beginning and developing of AS well. The study on drug's effect in treating AS was at its beginning. Until now, no such preventive and therapeutic drugs by directly interfere in AS were produced in China.Because of the complicated causes and uncertain mechnisms of AS, the multifactorial treatment was needed. The AS can be relieved by inhibiting some important steps in its formation. In this study, the SL extracts was used at early stage and maintained in the AS rabbit inflammatory reaction model, its effects on the inflammatory cells activation, proliferation, and on imflammatory mediators such as adhesion factor, cytokine, growth factor, chemotatic factor releasing were observed. Then the effects of AS such as size and extent were assessed to reflect the target effects and mechanisms of SL extracts. On the other hand, Considering the reports of monocytes function in AS and highβ2 integrin CD11/CD18 expression on leucocytes, we used flow cytometry(FCM) and Western blot in detection of CD18 changes in AS formation and verificated the persistent regular high CD18 expression on monocyte and the effects of SL.1. The effects of SL extracts on vessel pathological changes in AS rabbit modelAS rabbit model was made by Foley's tube endothelial impairment surgery and high lipid prey method. Oil red O stain of thoracic aorta, HE stain of femoral artery, toluidine blue stain of labrocyte, smooth muscle cell count were applied to observe the AS end point effect. The area of AS plaue in the model's thoracic aorta increased continuely from 6w. The femoral artery MIT value increased too. Denudation of local endotilial cells, infiltrating of inflammatory corpuscle and degeneration of elastic fibers of femoral artery were seen in HE stain in post-operative 1w. Amounts of bipolar lipoid vacuoles in theca interna and tunica media 5-6 muscular layers, smooth muscular cells translocation and proliferation were seen in the post-operative 6w. The thickened theca interna, bulged plaque inside which extracellular lipid and disrupted cells deposited , the theca interna and externa infiltrated wth inflammatory cells were found in 10w. The plaque area, lipid deposit, labrocyte infiltration, smooth muscular cells proliferation, MIT were decreased obviously in the Simvastatin group and in SL 1.12g/kg,2.24g/kg,4.48g/kg three groups.2. The effect of SL extracts on lipid metabolism in AS rabbit modelThe serum TC, TG, LDL-C, HDL-C were detected in 1w, 6w, 10w by the method of enzyme chromometry and phosphotungstic acid-magnesium precipitation. TC, TG, LDL-C increased continuely and were 6.8, 3, 8.8 times in 10w. HDL-C decreased.In Simvastatin 0.47mg/kg group, TC, LDL-C increased obviously and HDL-C were affected too. The TC,TG, LDL-C in SL 1.12g/kg, 2.24g/kg and 4.48g/kg groups were suppressed significantly. 3 .The effect of SL extracts on inflammatory factors in AS rabbit modelTNFα, IL-1β, IL-8 and CRP were detected in 1w, 6w, 10w by the method of radioimmunity and immunoturbidimetry. TNF increased a lot in every time point; IL-8 increased significantly in 1w and reached its peak in 6w, but decreased from 10w. CRP increased in 6w, 10w. IL-1βlevel was steady. TNFa,IL-8, CRP were suppressed in Simvastatin 0.47mg/kg, SL 1.12g/kg, 2.24g/kg, 4.48g/kg groups.4. The effect of SL extracts on adhesion factor, growth factor,chemotatic factor in vessel lesion of rabbit modelICAM -1,VCAM-1,MCP-1,IGF-1 were detected in 1w, 6w, 10w by the method of ELISA. ICAM-1, VCAM-I, IGF-1 increased obviously in every time point. MCP-1 increased significantly in 1w. VCAM-1, IGF-1 and MCP-lwere suppressed in Simvastatin 0.47mg/kg, SL 1.12g/kg , 2.24g/kg and 4.48g/kg groups.5. The effect of SL extracts on CD18 expressionExpression of CD18 on peripheral blood monouclear cells (PBMC) and its changing regularity was detected in 1w, 6w, 10w by the method of FCM. The CD18 highly expressed and reached peak in 1w, then decreased but still more than blank group in 6w and 10w. CD18 expression was suppressed in Simvastatin 0.47mg/kg group ,SL extracts 1.12g/kg, 2.24g/kg, 4.48g/kg groups.CD18 expression in vessel lesion of AS rabbit model was detected by Western blot and was found express significantly in the mature plaque. The suppression of CD18 had no statistical significance in Simvastatin 0.47mg/kg group, but was obvious in SL 1.12g/kg, 2.24g/kg, 4.48g/kg groups.6. The effect of SL extracts on THP-1,HUVEC adhesionTHP-1, HUVEC was cultured and used to measure the tolerance dose of SL extracts. The terminal experimental dose was 55 ul, 33ul, 20ul, 12ul, 7ul respectively. The serum drug content was 6.00 g/kg, 3.60 g/kg, 2.16 g/kg, 1.29 g/kg and 0.78 g/kg respectively with the interval of 60%, the 2.16 g/kg was equal to clinical equivalent dose.The coculture of THP-1 and HUVEC was stimulated with 5-40ng/ml TNFα, and the adhesion of cells was observed. Finally, 10ng/ml TNFα(10ng/ml) was selected as the adhesion inductor. Coomassie brilliant blue stain was used to detect the adhesive protein. THP-1 and HUVEC were suppressed dose dependently in the SL1, SL2, SL3 groups. 7. The effect of SL extracts on THP-1 liposuctionTC was detected by the method of enzyme chromometry and was found decrease dose dependently in the SL1, SL2, SL3, SL4, SL5 groups.SL extracts was an effect therapy to treat AS with the combination of Chinese traditional medicine theory, the damage-reaction theory, inflammation theory and pharmacology achievements. Our lab's former experiments found that SL could inhibit rat's AS formation by regulating lipid metabolism, inducing endothelial cells contraction, excretion and improving hemorrheology.Our latest experiment suggested that SL preventive administration decreased the AS end point effect .The anti-AS pathway in rabbit model was as follows: protect blood vessel endothelium and suppressed the monocytes, lympocytes, labrocytes adhesion and smooth muscular cells proliferation; improved lipid metabolism, decreased lipid deposit directly and inhibited lipid in anti-inflammation process indirectly; inhibited the produce and release of inflammatory mediators,decreased the adhesion factors, chemotactic factors and growth factor; The high CD18 expression was suppressed by SL in molecular and protein levels as proved by FCM and Western blot which maybe the comprehensive mechanism of SL effects. All those suggested the good integrity of SL in treating AS. SL extracts could inhibit the inflammatory reaction in initiation and development of AS .