Chinese medicine qishenjiangzhi decoction on atherosclerosis in rats
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Objective:To study the effect of Chinese medicine “Qishenjiangzhi decoction”(QSJZD) on atherosclerosis (AS) and to elucidate its mechanism further. Method:Twenty-four healthy male SD rats were randomly divided into 3 groups:control group, model group ,Chinese medicine treated group, The model rats of atherosclerosis were established by feeding with high cholesterol diet and injecting with vitamin D_2 for 6 weeks. Six weeks later, serum lipid levels of MDA, SOD and lipid were measured for all the rats. HE staining was used to study the pathology change of atherosclerosis. Immunohistochemistry and gelatin zymography were used to detect the expression and activity of MMP-2 in aorta vessel. Result:Comparing the chinese medicine treated group with model group, the serum level of TC, LDL-C and MDA were significantly reduced (P0.01) , while serum SOD level was obviously increased (P0.01) .The damage of the aortas in medicine treated group was less severe than that in model group. The expression and activity level of MMP-2 in medicine treated groups was significantly higher than the model groups. (P0.01). Conclusion:QSJZD could intervene the formation of atherosclerosis, The mechanism might be through suppressing TC, LDL-C and lipid peroxidation, and affecting the activity and expression of MMP - 2.Keywords:
Decoction
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Objective:To test the therapeutic effect of Taohong Siwu(THSW)decoction,a compound traditional Chinese herbal medicine:on Fas mRNA expression of hyperhomocysteinemia rats' aortae.Methods:Forty male SD rats were randomly divided into 4 groups with 10 in each group:untreated group,model group,THSW group and Folic Acid group.Thirty male SD rats received a high methionine diet to induce hyperhomocysteine and endothelial dysfunction.Rats in THSW group and Folic acid group were fed with THSW 4.59 g/(kg.d),Folic Acid 1.35 mg/(kg.d) respectively.Rats in untreated group and model group were fed with distilled water 2 mL/d intragastrically.At the end of 12 weeks,serum homocysteine levels were test by enzyme-linked immunosorbent assay(ELISA),plasma TNF-α and MCP-1 levels were tested with commercial kits.Fas,TNF-αmRNA were determined by RT-PCR.The expressions of cleaved Caspase3 was assayed by Immunohistochemistry.The expression of TNF-αof aortae were determined by western blot.Results:Compared with untreated group,Homocysteine in model group was obviously higher.The homocysteine levels in Folic Acid group were decreased obviously.Compared with untreated group,the levels of TNF-αand MCP-1 in model group in serum were higher.The TNF-αlevel in serum was lower than that in THSW group significantly.In aorta,the mRNA levels of TNF-αand Fas in model group were significantly heightened,THSW decoction can down-regulate the levels of aortic TNF-α mRNA significantly.The expression of cleaved Caspase 3 and TNF-αin model group were increased significantly compared with those in untreated group.However,the expression of cleaved Caspase 3 was downregulated in the THSW group.Conclusions:A high methionine diet can not only heighten serum homocysteinemia significantly but also induce the Fas mediated apoptosis.THSW decoction can down-regulate the expression of TNF-αmRNA in serum and reduce the aortic level of TNF-αat protein and gene level.
Hyperhomocysteinemia
Decoction
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Objective To observe the effect of the compound Yincha capsules(YC) on experimental atherosclerosis rabbits and explore its possible mechanism.Methods Twenty-five New-Zealand male rabbits were randomly divided into five groups,including normal control group,model group,high-dose YC-treated group,low-dose YC-treated-group and simvastatin group.Atherosclerotic models were established by feeding high fat diet for ten week.Levels of TC,TG,LDL-C,HDL-C,SOD and MDA in serum were tested.The plaque areas of aorta and pathomorphological changes were determined by image analysis.The expression of NF-κB and MMP-2 in aorta was examined with immunohistochemistry.Results YC significantly decreased the levels of TC,TG,LDL-C and MDA,but increased the levels of HDL-C,SOD.The area ratio of plaque of aorta in the treatment group reduced obviously compared with those of model group(P0.05).Degrees of AS formation in aorta of all YC groups were less than that in model group.The expression of NF-κB and MMP-2 in treatment group was significantly lower than those in control group.Conclusion YC can inhibit the formation of experimental atherosclerosis in rabbits,which may be related to decreasing blood lipid,anti-lipid peroxidation and the expression of NF-κB and MMP-2.
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To observe anti-atherosclerotic effect of Xuefu Zhuyu Granule (XZU) and Danlou Tablet (DT) on blood lipids, platelet derived growth factor (PDGF), vascular smooth muscle cells (VSMCs) proliferation, extracellular signal-regulated kinase (ERK) signal pathway in atherosclerosis (AS) model rats, and to explore their potential mechanisms.Forty male Wistar rats were randomly divided into five groups, i.e., the normal control group, the model group, the Atorvastatin group, the DT group, the XZG group, 8 in each group. Rats in the normal control group were fed with basic forage for 12 weeks, while rats in the other four groups were fed with high fat forage plus intraperitoneal injection of vitamin D3 to build AS model. Then rats in the model control group, the Atorvastatin group, the DT group, the XZG group were administered with normal saline, Atorvastatin suspension (0.18 mg/mL), DT suspension (45 mg/mL), and XZG (1 g/mL) by gastrogavage for 8 successive weeks, respectively. After intervention serum levels of TC, TG, LDL-C, HDL-C, and PDGF were detected by ELISA. Pathological changes in thoracic aorta were observed by HE staining. Protein expression levels of ERK1/2 and pERK1/2 in thoracic aorta were measured by Western blot.Compared with the normal group, serum TC, TG, LDL-C, PDGF levels, and expression levels of ERK1/2 and pERK1/2 significantly increased (P <0. 05) in the model control group. HE staining showed irregular intimal thickness, accumulated endothelial foam cells, lipids deposited, disarranged media VSMCs, forming typical AS plaque. Compared with the model group, TC and PDGF levels decreased in all medicated groups (P < 0.05, P < 0.01). Serum levels of TG and LDL-C significantly decreased in the Atorvastatin group and the DT group (P < 0.01, P < 0.05). Expressions of ERK1/2 and pERK1/2 significantly decreased in the Atorvastatin group, the DT group, and the XZG group (P < 0.01). HE staining also showed typical AS plaque in three medicated groups, but with reduced pathological degree of endometrial hyperplasia and plaque area.XZG and DT could reduce the plaque area and attenuate pathological degree of AS in model rats, thereby postponing the progress of AS. Its mechanism might be achieved through reducing serum lipids and release of PDGF, inhibiting ERK signal pathway activation and VSMC proliferation.
Intraperitoneal injection
Thoracic aorta
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Objective To explore the effect of silybin-phospholipid complex treatment on fatty liver disease and atherosclerosis in rats fed with high-fat diet.Methods Twenty-two SD rats were randomly divided into three groups: the control group(n=6),the model group(n=8) and the therapeutic group(n=8).The control group were fed with normal diet,and the other two groups were fed with high-fat diet plus Vitamin D and PTU.Meanwhile,the therapeutic group were treated with silybin Capsules(225 mg·kg-1·d-1).All rats were sacrificed at the end of the 16th week of the experiment.The pathological changes in livers and aortas in all rats were pathologically evaluated with HE stain,and the serum biochemistry index was also measured.Results Significant hepatic steatosis,intra-lobule inflammation and spotty hepatocyte necrosis were seen in the model group.In addition,tunica media of aortas were slightly thickened and calcified plaque was observed under the endothelium in the model group.Compared with the model group,liver steatosis and inflammation were relieved in the therapeutic group pretreated with Silybin Capsules(P0.05),and the serum levels of ALT were also significant decreased(P0.01).However there was no difference on serum levels of fasting glucose and lipids,as well as lesions in aorta between the therapeutic group and the model group.Conclusion Silybin-phospholipid complex can improve the degree of steatohepatitis in rats induced by high-fat diet.However,it has no obvious benefit on hyperglycemia,dyslipidemia and aorta atherosclerosis
Steatosis
Therapeutic effect
Dyslipidemia
Steatohepatitis
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Objective To study the role of serum TNF-α level in formation of atherosclerotic plaques. Methods Ten male big ear rabbits were randomly divided into model group(group A, n=5) and normal control group(group B, n=5). After damage to abdominal aorta intima was induced with balloon, the animals in group A were fed with high fat diet for 15 weeks while those in group B were fed with normal diet for 15 weeks. Fasting blood was taken from the ear marginal vein to measure the serum levels of TC, TG, HDL-C, LDL-C and TNF-α before and by the end of week 15. The animals were killed. Their abdominal aorta was isolated and observed under optical microscope. Apoptosis was detected by TUNEL. Results The animal atherosclerosis model was successfully established. The serum levels of TC, TG, LDL-C and TNF-α were significantly higher in group B than in group A(P 0.05). The serum levels of TC, TG and LDL-C were positively related with the serum TNF-α level(P 0.05). However, the serum HDL-C level was not significantly related with the serum TNF-α. level. The serum TNF-α level was positively related with the apoptosis index in abdominal aorta plaques. Conclusion TNF-α may be involved in apoptosis in atherosclerotic plaques. Serum levels of TG, TC, LDL-C and TNF-α are the risk factor for atherosclerosis.
Abdominal aorta
Normal group
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Objective To observe the effects of Chinese herbs on lipid peroxidation reaction in rats with non-alcoholic fatty liver(NAFL),to inquire into its prevention and treatment mechanism.Methods Thirty-two male SD rats were randomly allocated into 4 groups: control group,model group,Dongbao Gantai(DBG) group and Ganshutang decoction(GSTD) group,8 in each.All groups except the control were fed with an 8-week high fat diet to prepare NAFL models.General condition,pathological changes of liver tissues were observed,and blood lipids and the level of tumor necrosis factor-alpha(TNF-α)were detected,so were the levels of MDA,SOD of liver tissues.Results In model group,teatosis appeared in liver tissues which were gradually infiltrated with inflammation cells,and necrosis was present in hepatocyte,percolate in portal area.There was significant difference in the scores of inflammation activity between model and control groups(P0.05),model group was significantly different from control and GSTD groups(P0.05),and there was significant difference between DBG and GSTD groups(P0.05).There was significant difference in TG、LDL-C、HDL-C between 4 groups(P0.05),so was there between model and control groups(P0.05),and significant difference was seen in levels of TG、LDL-C between DBG and model groups(P0.05);There was significant difference in serum levels of TG、LDL-C、HDL-C between DBG and model groups(P0.05)and there was significant difference in serum levels of LDL-C、HDL-C between DBG and GSTD groups(P0.05).In model group,the levels of TNF-α,MDA in liver tissues obviously increased,while the active levels of SOD obviously decreased.As comparing with model group,GSTD could improve the liver pathology structure,and play a role in decreasing significantly blood lipids,the level of TNF-α and liver tissue MDA.Conclusion Ganshutang decoction can prevent and treat NAFL to a certain degree,the mechanism of which is perhaps related to prevention of lipid peroxidation and decrease of the infiltration of inflamed cytokine.
Decoction
Therapeutic effect
Alcoholic fatty liver
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Objective To investigate the effect of pioglitazone on the expression of connective tissue growth factor(CTGF)and matrix metalloproteinase 9(MMP-9)in rats fed with high-fat diet.Methods Twenty six male Sprague Dawley rats were randomly divided into control group(n=9)and high-fat diet group(n=17).After fed with hypercholesterol diet for 12 weeks,the rats of high-fat diet group were randomly subdivided into model group(n=8)and pioglitazone group(n=9).The levels of total cholesterol(TC),triglycerides(TG)and low-density lipoprotein cholesterol(LDL-C)in high-fat diet group were significantly higher than those in control group(all P0.05),suggesting the modeling was successful.After 4 weeks of treatment,serum lipid level was measured by enzyme-linked immunosorbent assay,blood glucose was measured by automatic biochemical analyzer,and CTGF and MMP-9expression were detected by immunohistochemistry.Aortic histomorphological changes were observed.Results Serum TC and TG levels in pioglitazone group were significantly lower than those in model group(both P0.05).Aortic endothelial tissue was largely intact,occasionally dropped,proliferation of smooth muscle cells were not obvious.The average optical density of CTGF and MMP-9in model group were significantly higher than those in control group and pioglitazone group(all P0.05),but there was no significant difference in the optical density between pioglitazone group and control group(both P0.05).Conclusion Hyperlipemia can induce up-regulation of CTGF and MMP-9expression in aorta of rats.Pioglitazone can inhibit the expression of CTGF and MMP-9,and effectively delay the development of atherosclerosis.
Pioglitazone
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Objective: To observe the ef ect of TongmafiHuazhuo Decoction(TMHZ) on mRNA and protein expression of ApoAⅠ, B in the liver of model rat with coronary heart disease, and to explore its molecular mechanism in treatment of coronary heart disease. Methods: Totally 66 clean grade SD rats were randomly divided into a normal group, model group, Xuexhikang(XZK) group, TMHZ high, medium and low dose group by weight and sex. Except for the normal group, rat models with coronary heart disease were established by using lipid infiltration calcium overload, and the lead Ⅱ ECG of the rats was recorded to test whether it was successful. After intragastric administration for 4 weeks, the rats in each group were routinely sacrificed to extract tissues. The liver mRNA and protein expression of ApoAⅠ, B were measured by RT-PCR and Western Blot, and then their expression differences were analyzed. Results: ①The mRNA and protein expression levels of ApoAI in the model group were obviously lower than those in the normal group(P0.05). Compared with the model group, mRNA and protein expression levels of ApoAI in each TMHZ treatment group and XZK group were significantly increased(P0.05, P0.01). ②The mRNA and protein expression levels of ApoB in the model group were remarkably higher than those of the normal group(P0.05, P0.01), mRNA and protein expression levels of ApoB in each TMHZ treatment group and XZK group were significantly lower than those of the model group(P0.01, P0.05). Conclusion: TMHZ is able to increase or stabilize ApoAI expression, and decrease or inhibit ApoB expression in gene and protein levels, thus increase the content of ApoAI and lower ApoB in plasma, which may be its molecular mechanisms to treat coronary heart disease, lower blood lipids, and delay the process of atherosclerosis.
Decoction
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Objective In this study,the effect of shuganhuoxue(SGHXT) on Insulin-like growth factor-1(IGF-1) expression of vessel wall was explored with immunity histochemistry.Method 32 healthy rabbits were randomly divided into four groups:Normal control group(A),Model group(B),Fluvastatin group(C) and SGHXT group(D).The four groups were given ordinary feed,high fat diet,high fat diet with Fluvastatin capsules,high fat diet with SGHXT.After 5 weeks,group B、C and D were given Right abdomen-iliac artery balloon injury of endometrial.After 7 weeks,group A,B,C and D were collected blood samples,tested lipid and explored Insulin-like growth factor-1(IGF-1) expression of vessel wall with immunity histochemistry.Result The results showed that the IGF-1 expression in model group was enhanced than that in the normal group.However,the expression in SGHXT group decreased less than that in the model group.Conclusion It suggested that one of the effect of SGHXT on preventing VSMC proliferation was its influence on IGF-1 mRNA expression level.
Fluvastatin
Decoction
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Objective
To investigate the effect and related mechanism of Zinimaibi Decoction on the expression of lectin-like oxidized low-density lipoprotein receptor (LOX)-1 in the hindlimb arteries of diabetic rats.
Methods
A total of sixty Sprauge-Dawley rats were included in this study. Ten rats were randomly selected as normal control group according to random number table method, whereas all the other rats were fed with high-glucose and high-fat diet combined with the injection of streptozotocin (25 mg/kg) from tail vein to induce diabetes. After successful modeling, all diabetic rats were randomly divided into non-drug intervention group, cilostazol group(10 mg·kg-1·d-1), low dose of Maibi Decoction group(12 g·kg-1·d-1), medium dose of Maibi Decoction group(24 g·kg-1·d-1)and high dose of Maibi Decoction group(48 g·kg-1·d-1)according to random number table method, with 10 rats in each group. After 4 weeks of intervention, body weight, blood glucose, liver function, renal function and total cholesterol(CHO), triglyceride(TG), high density lipoprotein-cholesterol(HDL-C) and low density lipoprotein-cholesterol(LDL-C) were examined.HE staining was used to observe the morphilogical changes of hindlimb artery intima. Western blotting was used to analyze the expression of LOX-1, nuclear factor-κB(NF-κB) p65 and vascular cell adhesion molecule(VCAM)-1 of hindlimb.
Results
The weight of diabetic rats were decreased and blood glucose were increased significantly compared with non-diabetic rats after modeling (F=4.63, 120.27, all P 0.05). The serum CHO and LDL-C level in the medium and high dose of Maibi Decoction group were downregulated compared with non-drug intervention group, the serum TG level in normal control group, cilostazol group, and all Maibi Decoction groups were decreased significantly compared with non-drug intervention group(F=4.10, 4.97, 17.23, all P<0.01). Mucoid degeneration were found in the hindlimb artery in non-drug intervention group but were improved in all intervention groups, especially in the medium and high dose of Maibi Decoction groups. Compared with non-drug intervention group, the expression of LOX-1 and VCAM-1 protein were decreased significantly in low, medium, high dose of Maibi Decotion intervention group and cilostazol group (F=13.25, 43.01, all P<0.01). The protein level of NF-κB p65 were also decreased significantly in cilostazol group and high dose of Maibi Decoction group compared with non-drug intervention group (F=13.83, P<0.01).
Conclusions
Zinimaibi Decoction downregulates the expression of LOX-1 in the hindlimb arteries of diabetic rats and inhibits the expression of VCAM-1 by decreasing the expression of NF-κB p65.
Key words:
Zinimaibi Decoction; Diabetic lower limb atherosclerosis; Lectin-like oxidized low-density lipoprotein receptor-1; Nuclear factor-κB p65; Vascular cell adhesion molecule-1
Hindlimb
Decoction
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