Study Of Alismatis Rhizoma On Auti-hyperlipidemia In Vitro

Objective:The effects of the serum containing active ingredients of Alismatis Rhizoma on the viability,cycle,and apoptosis of vascular endothelial cells treated with ox-LDL oxidation and cytokine(IL-1 and TNF-alpha),NO and NOS,antioxidant indicators(SOD,MDA,)as well as related genes and signaling pathways protein(Bcl-2,Akt and Caspase-3)indicators of the cells were characterized by cell culture,CCK-8 method,flow cytometry,ELISA,RT-PCR,Western-blot,etc.According to the differences between the drug group and model group,the protection mechanism of Alismatis Rhizoma on the vascular endothelial cells was concluded;further,the possible mechanism on anti-hyperlipidemia was discussed.The obtained results provided experimental basis for the clinical use of Alismatis Rhizoma.Research Methods:(1)the method of CCK-8 was used to observe the viability of the cells treated with the different doses of the serum containing active ingredients of Alismatis Rhizoma to obtain an effective dose.(2)the fluorescence analysis technique and flow cytometry were used to characterize the cell cycle and apoptosis of the cells treated with different doses.(3)the kits were used to assay antioxidant indicators(SOD,MDA)and NO,NOS expression.(4)ELISA was used to assay cytokines expression(IL-1,TNF-alpha).(5)RT-PCR and Western-blot were used to assay the expression of Bcl-2,Akt and Caspase-3.Results:based on the results of serum group and ox-LDL-induced model group,the serum containing active ingredients of Alismatis Rhizoma can improve vascular endothelial cell morphology and promote cell proliferation.The results of flow cytometry and Hoechest detection showed that drug group apoptosis rate was lower significantly.The results of kits detection showed that the activities of SOD and NOS of serum group were higher than those of ox-LDL-induced model group significantly,and that vascular endothelial cells the ability of secreting NO of vascular endothelial cells also increased after treated with the serum.The results of RT-PCR proved that Bcl-2 gene expression of each serum group was up-regulated significantly and active caspase-3 gene expression was down-regulated significantly.The results of Western-blot showed that Bcl-2,caspase-3 and Akt protein expression were up-regulated significantly.Conclusion:Alismatis Rhizoma can protect vascular endothelial cells damaged by ox-LDL such as improving cell morphology,promoting cell proliferation,inhibiting apoptosis,enhancing the activity of SOD and NOS,and improving the ability of secreting NO.Various doses of the serum can inhibit apoptosis of the cell damaged by ox-LDL by up-regulating the expression of Bcl-2,Caspase-3,and Akt protein,reducing activation of caspase-3 gene expression.Our results indicated that Alismatis Rhizoma can protect vascular endothelial cells damaged by ox-LDL to maintain its normal structure and functionHere,in vitro studies were carried out to investigate the nature of hyperlipidemia caused by ox-LDL,and to explain the treatment mechanism of Alisma on hyperlipidemia.Experimental subjects are vascular endothelial cells;experimental drug is the serum containing active ingredients of Alismatis Rhizoma.The techniques of cell biology and molecular biology were used to verify the protection mechanism of Alismatis Rhizoma on endothelial cells.We speculated that protection of endothelial cells of Alismatis Rhizoma may be one of the ways of hyperlipidemia treatment.Our study has some theoretical and practical significance.