| OjectiveAtherosclerosis (AS) is one of the main diseases that harm human health. Researches on the pathogenesis of AS are always hot spot. Although many theories have been proposed, such as the proliferation of tunica media's smooth muscle multiplication theory, the thrombus source theory and so on, neither of them can provide satisfaction explanation. In 1999 Ross has proposed the concept of "AS is one kind of inflammation disease", claimed that AS has the chronic inflammation response character in the pathology process, and its developing process throughout the inflammation response. In recent years, many researches believe that AS was related to the infection of Chlamydia pneumoniae(Cpn), our topic group once reported that TNF-alpha(TNF-α) is increased in the supernatant of the blood vessel endothelial cell(VEC) induced by Cpn. Baicalin of high-dose, middle-dose and low-dose all cause it decline. On the foundation of early research, we continued to observe the influence of baicalin to the VEC induced by TNF-α, for further discussing baicalin's anti-damage mechanism.Method1. The culture of human umbilical vein endothelial cells (HUVEC)2. TNF-αand baicalin jointly to the proliferation of HUVEC3. Examine CD54 and CD106 expression on HUVEC superficial by flow cytometry(FCM)4. Examine CD106 expression on HUVEC superficial by Lipopolysaccharide (LPS) and Phorbol myristate aceta (PMA) 5. TNF-αinduces HUVEC's apoptosis6. Detect HUVEC's apoptosis by FCM7. Statistical methodUses the variance analysis (ANOVA) of SPSS11.0 software package to carry on the data analysis, during the multi-groups comparisons uses the LSD.Inhibition ratio=(1-experimental group/comparison group)×100%Results1. The normal HUVEC under the inverted microscope are adherent cells, to assume the triangle, the polygon, or fusiform shape. It's like "pavingstone". After infected by TNF-α, the cellular form have not seen obviously changes.2. TNF-αstimulates 24h the HUVEC cells multiplication, comparing with normal group it has the tendency of reduce, but non-statistics difference. Baicalin of high dosage (120μg/ml) the group have no influence to normal cells multiplication, and baicalin of the high dosage may have the function of improving TNF-αinhibitory action, promoting the HUVEC proliferation. It is 16% growth rate, also P＜0.053. Stimulated by TNF-αfor 24h, CD54 which expressed on the HUVEC's surface is increased approximately by 1.6times. Baicalin of high dosage (120μg/ml) have no influence to normal cell's CD54. The high and middle dosage groups can effectively reduce the expression which is induced by TNF-α. The inhibition ratios are respectively 41%, 15%. TNF-αdoes not obviously affect the CD106 of HUVEC surface.4. After LPS and PMA stimulating 24h, the HUVEC surface's expression of CD106 are no difference comparing with the normal group.5. TNF-αmay induce HUVEC to apoptosis, when the density is 100 ng/ml, at 24h, the apoptosis rate is the highest, and it's approximately 2.4 times to normal group.6. Baicalin of high dosage may suppress the rate about of 38% increased by TNF-α.ConclusionAfter stimulated by TNF-α, CD54 of the HUVEC increases. Meanwhile the apoptosis rate also obviously goes up, but does no affection to the CD106. These suggest that TNF-αdamages the endothelial cells by two ways, one hand it makes the membrane surface's attaches the factor (CD54) increase, and on the other hand may direct induce cells to apoptosis. Baicalin has a certain inhibitory action to the expression of CD54. It can also reduce the apoptosis rate. Both of them manifested that baicalin has the function of anti-inflammation and anti-apoptosis.Our researches from two aspects of adhesion factor and apoptosis, using FCM and MTT, has discussed TNF-αto the HUVEC's damage function. The results contribute to the theory of "AS is a chronic inflammation" as the cytology experiment.Some studies are needed to find out the molecular way of baicalin reducing the expression of adhesion factor and the signal pathway which baicalin suppress the apoptosis of endothelial cells.
|
PDF Full Text Request