| Objective To research the protective effect of qing mai yin and its separated recipes containing serum on the inferior mesenteric artery endothelial cell(Ealy926) and related factors induced by tumor necrosis factor-α(TNF-α).Methods Preparation of medicated serum,On the EAhy-926 cells were incubated in vitro and divided into 9 groups of normal control group, the10% low dose qing fa group, the 10% normal dose qing fa group,the 10% high dose qing fa group,huo xue group、quan fang group, ruan jian group, medicine group, TNF-α injury group. etc. The serum containing medicine and different concentrations of qing fa serum containing handle with the TNF-α endothelial cells 24 hours after, morphological changes of EAhy-926 cells were observed under light microscope; cell organelles change of EAhy-926 cells were observed under the electron microscope,CRP,IL-6,ET-1,VEGF was detected immunohistochemistry and watchted their expression in cells under light microscope,Nitrate reduction assay NO, color depth determined by the level of concentration,NF-κB gene expression was detected by Real-time PCR.Results ①The changes in general morphological cells under Light microscope Compared with TNF-α injury group,the serum containing medicine was significantly increased the cell viability, Each group damage less and cell number, cells of huo xue group are relatively few,②The changes in the structure of cells under electron microscope.The damage of compared with TNF-α injury group,the serum containing medicine was significantly pure TNF alpha cells of small injury group. The cell organelles was least damage in qing fa group, ruan jian group, medicine group and the cell ultrastructure was no significant difference compared with normal control group,③ NO concentration and the expression of CRP、IL-6、ET-1 were reduced,Multiple comparison between groups,the 10% normal dose qing fa group has a good effect than other groups in lower ET-1;medicine group was best, the 10% normal dose qing fa group is better than ruan jian group and huo xue group to reduce concentration of NO and the expression of CRP,IL- 6, VEGF,comparison among different concentration groups of qing fa group, the 10% normal dose qing fa group than other best concentration effect ④ The expression of NF-κB gene were reduced,Multiple comparison between groups, the 10% normal dose qing fa group,quan fang group and medicine group, has advantages in control on the expression of NF-κB, the effect of medicine group was best, the 10% normal dose qing fa group,quan fang group, the 10% low dose qing fa group, ruan jian group huo xue group followed in turn. The 10% normal dose qing fa group,quan fang group fairly and below huo xue group(P < 0.05), shows that effect of the 10% normal dose qing fa group is better than huo xue group. Qing fa comparison among different concentration group, the 10% normal dose qing fa group has the best effect than other concentration in ruduceing NF-κB gene expression.Conclusion:① qing mai yin and its separated recipes containing serum Possible injury by antagonizing TNF-α on cultured arterial endothelial cells to maintain its structural integrity; To serve to prevent or mitigate the effect of arterial endothelial cell injuried,The protection effect of medicine group and the qing fa drug-containing serum were best② Qing mai yin and its separated recipes containing serum could ruduce the damaged endothelial cells within the NO concentration,and the expression of CRP,IL-6,ET-1,VEGF to prevent any further damage to endothelial cells, effect of the 10% normal dose qing fa group is better than huo xue group ③Qing mai yin and its separated recipes containing serum could by blocking the TNF-a activated the NF-κB pathway, classic regulation so as to reduce the NF-κB gene expression of vascular endothelial inflammatory response.④From all angles of the experimental observation, the effects of the 10% normal dose qing fa group was better than that huo xue group in the qing mai yin and its separated recipes. |
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