Prophylaxis Action And Machenism Of Detoxicating Decoction Of Coptis On The Pathological Changes Of Aorta Induced By Hyperlipidemia

The pathogenesis of atherosclerosis is very complex.It is known that endothelium dysfunction induced by hyperlipidemia is the initiation link in atherosclerosis.Especially, if the LDL was oxidatively modified to ox-LDL by oxidant released from endothelial cell and macrophage,ox-LDL plays an important role in the pathogenesis of atherosclerosis.Detoxicating Decoction of Coptis（DDC,Huanglian Jiedu Tang） is a famous prescription of TCM,has the functions of clearing away heat and detoxicating, which is composed of Coptis Root（3g）,Scutellaria Root（6g）,Phellodendron Bark（6g） and Capejasmine Fruit（9g）.The major chemical components are Berberine,Baicalin and Gardenoside.Their actions include both regulating fat metabolism and anti-inflammation.Therefore it is interesting to explore the anti-atherosclerosis effect and mechanism of DDC.In this paper,experiment will be carried out on male Sprague-Dawley Rat and Human Vascular Endothelial Cell（HUVEC）.Part 1 Research on the effect of DDC of blood lipids lowering in male SD ratsMethods:The experimental hyperlipidemia model of SD rat was induced by High Fat Diet （HFD）,which was composed of cholesterol（2%）,sodium cholic acid（0.8%）, propylthiouracil（0.1%）,and etc..DDC is composed of Coptis Root（3g）,Scutellaria Root （6g）,Phellodendron Bark（6g） and Capejasmine Fruit（9g）.At the 53^th day serum lipids including TC,TG,LDL-C and HDL-C were measured to diagnose experimental hyperlipidemia.At the 115^th day,body weight,serum lipids including TC,TG,LDL-C and HDL-C were measured.Results show that HFD could lower the body weight of rats, heighten the content of serum TC,LDL-C.It shows that DDC could heighten body weight,lower serum TC,LDL-C,glucose,and MDA of experimental hyperlipidemia rats. Part 2 Research on the prophylaxis action of DDC on the pathological changes of aorta after hyperlipidemia in male SD rats.Methods:When experimental hyperlipidemia was forming in rat at the 53^th day, beginning inject DDC into rat for 61 days.At the 115^th day,HE staining and SudanⅣfat staining were used to investigate the histo-pathological changes of aorta.Immunohistochemical technique was used to check express of Smooth Muscle Actin in Smooth Muscle Cells in aorta walls.Cell cycle and apoptosis of vascular endothelium cells in tunica intima of aorta was measured with Flow Cytometer（FCM） and analyzed.And transmission electron microscope（TEM） was utilized to observe ultrastructure changes in aorta walls.Results show that HFD could changes the aorta morphology and function.By HE staining,the morphological changes of aorta were found that the tensity was increasing,the tunica intima was thickening,the tunica media was thinner,the elastic membrane in tunica media was tensed.By SudanⅣstaining,a thin lipid layer was found on vascular endothelium cells layer.After immunohistochemical staining and analyzing it was found that the Actin of vascular smooth cell in tunica media was increasing.The ultrastructure changes were showed that vascular endothelium cells were falling off the tunica intima and hurt,their nucleus were larger,they were proliferating.Splits were found in the internal elastic membrane.Cell cycle analysis found the proliferous index（PI） of VEC was promoted.And biochemistry test point out concentration of NO and iNOS in serum were enlarging.It shows that DDC could improve the morphology and function changes of aorta stated as above.Part 3 Research on the other related actions of DDC in the experimental hyperlipidemia male SD rats.Methods:At the 115^th day,serum glucose,TNF-α,IL-6,IL-1β,ALT and AST were measured.SudanⅣfat staining was used to investigate the histopathological changes of liver.Results show that HFD heighten the content of serum TNF-α,IL-6 and ALT.Results also show that DDC could lower the content of serum glucose,TNF-αand IL-6.Part 4 Research on the effect of DDC serum on Human Vascular Endothelial CellMethods:DDC serum was extracted from male Wistar rats which were fed with DDC for 9 times in five days.Briefly,100μL cells(5×10⁸L^-1) were seeded in each well of 96 well micro-plate,after incubated in a CO₂ incubator at 37℃,the cells were treated with various medium. After that,HUVEC viability(A_570nm values) was determined by MTT method.Results showed that DDC serum could not protect HUVEC hurt by Ox-LDL.It also showed that DDC serum could block the multiplication of HUVEC promoted by FBS.CONCLUSIONAll of results suggest that HFD could induce rat to experimental hyperlipidemia.And during experimental hyperlipidemia period,the aorta function may change before fat lines forming in AS.Detoxicating Decoction of Coptis could prevent the morphology and function changes of aorta.The mechanism is that DDC could regulate fat metabolism, lower TC and LDL.It could lower MDA,so it has anti-oxidation action and block LDL oxidized.It also has anti-inflammation effect,because it lower NO,TNF-αand IL-6.It could block the extraordinary multiplication,proliferation and apoptosis of VEC.In sum, DDC has prophylaxis effect on the pathological changes of aorta induced by HFD.Maybe DDC could use to prevent atherosclerosis.