Effects Of The Extractions Of Polygoni Cuspidati Rhizoma Et Radix And Crataegi Fructus On As Vulnerable Plaque And Apoptosis Regulators’ Expressions

ObjectiveTo observe the effects of the extractions of traditional Chinese herbs:Hawthorn and Polygonum Cuspidatum (HPC) on blood lipid(triglyceride[TG],cholesterol[TC], low density lipop-rotein[LDL] and high density lipoprotein[HDL]) and pathology morphologic changes of vascular vulnerable plaques of the ApoE gene kick-out[ApoE (-/-)] mice model.To explore the effects of HPC on mRNA expressions of apoptosis regulators:Bcl-2Associated X(Bax),B-cell leukemia-2(Bcl-2),cysteinyl aspartate specific proteinase-9(Caspase-9) of the artery.To provide experimental data for HPC used for prevention and treatment of atherosclerosis(AS).Methods64ApoE (-/-) mice (8weeks old, male) were fed with a high-fat diet for13weeks and then randomly chose4mice, removed their arteries and confermed the formation of vulnerable plaques.The rest of ApoE (-/-) mice were randomly divided into5groups:the model group,the simvastatin group,the HPC high-dose group,the HPC medium-dose group,the HPC low-dose group,except the C57control group,each group had12mice.12C57BL/6J mice (8weeks old,male)which had same heredity background with ApoE (-/-) mice and were fed with ordinary diet were selected into C57control group.The C57control group and the model group were given equal-volume distilled water orally,others were given Hawthorn extr-actions(86.7mg/kg) and Polygonum Cuspidatum extractions(40mg/kg),Hawthorn extractions (173.4mg/kg) and Polygonum Cuspidatum extractions (80mg/kg),Hawthorn extractions (346.8mg/kg) and Polygonum Cuspidatum extractions (160mg/kg), simvastatin(2.6mg/kg) respectively.All the mice were administered once a day for13weeks.During the administation period,all the groups were fed with SPF standard high-fat diet except the C57control group.Finally,the blood lipid(TG,TC,LDL and HDL) was tested by blood collected from ophthalmic vessels.The pathologic morphology of arteries was tested by HE staining and electronic light microscope. And the expressions of Bcl-2,Bax,Caspase-9mRNA in arteries were detected by RT-PCR,the protein expressions of Bcl-2,Bax,Caspase-9in the plaque of arteries were detected by immunohistochemical staining. ResultsThe blood lipid test showed that:compared with the C57control group,the levels of TC,TG,LDL,HDL of the model group all increased,TC and LDL increased significantly(P<0.01).After13weeks,compared with the model group,the LDL level of the simvastatin group and the HPC low-dose group decreased obviously (P<0.05);the levels of LDL,TC,TG in the HPC high-dose group all decreased significantly(P<0.01or P<0.05);the HDL level increased obviously in the HPC medium-dose group and the HPC low-dose group(P<0.05or P<0.01);the HPC high-dose group had the strongest power to reduce the TQLDL and TC level(P<0.01or P<0.05);there was no significant difference between each medication group about HDL level(P>0.05).The pathologic morphology of the ApoE(-/-) artery indicated that in the C57control group,the artery wall was thin and its structure was clear and orderly. There was no foam cells or plaques at intima.In the model group,the intima was locally incrassate and protuberant into vessel,endothelial cells was injured,the integrality of intima was destroyed,vulnerable plaques formed,lots of foam cells,necrotic tissues and cholesterol crystals presented in the plaques;smooth muscle cells proliferated, swelled and arranged disorderly.After13weeks administration,all the pathology morphology changes of medication groups were alleviated compared with the model group.The size of the vulnerable plaques were decreased in all of the medication groups.In the HPC high-dose group,there were lots of foam cells in the plaque,no calcification.In the HPC medium-dose group,little atherosclerosis necrotic area and calcification could be seen in the plaque.There were little cholesterol crystal and medium atherosclerosis necrotic area could be seen in the plaque of HPC low-dose group.In the simvastatin group,there were lots of foam cells in the plaque and medium atherosclerosis necrotic area.RT-PCR showed that:Compared with the C57control group,both the expression of Bax and Caspase-9mRNA in arteries increased significantly(P<0.01)in the model group;the decrease of the expression of Bcl-2mRNA was no statistical difference(P>0.05).After13weeks’administration,compared with the model group,both the simvastatin group and the HPC high-dose group significantly decreased the Bax mRNA expression in arteries(P<0.01or P<0.05),and the simvastatin group was more powful; the Caspase-9mRNA expression in the HPC high-dose group and the simvastatin group decreased significantly(P<0.01or P<0.05); the Bcl-2mRNA expression in the HPC high-dose group,the HPC medium-dose group significantly increased(P<0.05).Immunohistochemical staining showed that:Compared with the C57control group,both the protein expression of Bax and Caspase-9in vulnerable plaques increased significantly(P<0.01) and Bal-2decreased significantly(P<0.05)in the model group.After13weeks’administ-ration,compared with the model group,all the protein expressions of Bax in vulnerable plaques of HPC high-dose group,HPC medium-dose group and the simvastatin group decreased significantly(P<0.01or P <0.05);and the simvastatin group and HPC medium-dose group both significantly decreased the Caspase-9protein expression in vulnerable plaques(P<0.01);the protein expression of Bcl-2in vulnerable plaques in HPC high-dose group,HPC medium-dose group and the simvastatin group increased significantly(P<0.01or P <0.05),the HPC high-dose group was the most significant;the Bax protein expression in the HPC high-dose group was the most significant(P<0.01).ConclusionHPC could improve blood lipid levels,protect vascular intima,alleviate the development of AS lesions in ApoE (-/-) mice apparently.It might inhibit AS progression through downregulating Bax,Caspase-9mRNA expression and up regulating Bcl-2mRNA expression,then regulating vascular cells’ apoptosis.