| Objective:(1)Observe the clinical effect of Fufangteng Yixin Decoction on Thoracic Obstruction due to Qi Deficiency Blood Stasis(chronic stable angina) and influence on its endogenous active substance level changes.(2)experimental study of Fufangteng Yixin Decoction on protective effect and mechanism of human endocardium microvascular endothelial cell(HUMCE) after impairment of hypoxia/reoxygenation(H/R) in vitro.Methods:(1)82 cases of Thoracic Obstruction due to Qi Deficiency Blood Stasis(chronic stable angina), at the age of 35 to 79, were enrolled for the experiment, and randomly divided into two groups, including the treatment group and the control group with 41 cases respectively. The control group was given the routine treatment of western medicine(including drugs for anti-platelet, Nitrates, beta-blocker, ACEI.). Based on the routine treatment, Fufangteng Yixin Decoction was given in the treatment group twice per day, with 200 ml for each time. During four-week treatment, the efficacies of TCM manifestations, angina, electrocardiogram as well as changes of serum VEGF, ET-1, hs-CRP and IL-1β for patients were observed and compared between treatment before and after.(2)By comparing the approaches of aqueous extractions and ethanol extraction Fufangteng Yixin Decoction and YixinDecoction, the one with higher extraction efficiency was used for extractions of both. The acute toxicity test of Fufangteng Yixin Decoction, Yixin Decoction and ethanol extracts on mice was observe, so as to evaluate the safety of those medicines. Applying for serum pharmacological methods, rat serums with Fufangteng Yixin Decoction and Yixin Decoction in different concentrations were prepared to intervene HUMCE. Moreover, MTT was utilized to detect the activity of HUMCE to choose the best rat serums with Fufangteng Yixin Decoction and Yixin Decoction. In the experiment, it was divided into the control group and modeling groups, including the model-control group, the Fufangteng Yixin Decoction group, the Yixin Decoction group and the positive-control group[captopril10-2mol/L(CAP1), 10-3mol/L(CAP and 10-4mol/L(CAP3)].After H/Rpost-injuried HUMCE model was established in vivo, the intervention of rat serums with medicines was applied in accordance with the experimental grouping: the activity of HUMCE was tested by MTT; postive cell counts of Caspase-3 of HUMCE was detected via flow cytometry; the content of NO and the activity of NOS in the supernate of HUMCE by nitrate reductase method; the content of VEGFR-2 and VEGFn in the supernate of HUMCE via ELISA; the protein expression of ET-1 and ECE of HUMC was determined by Western blot; the gene expression of ET-m RNA and i Nos-m RNA of HUMCE was tested by PCR.Results:(1)Curative effects comparison of TCM syndromes: The total effective rate of treatment group was 90.2%, the total effective rate of control group was 73.2%. The comparison of TCM syndromes of the 2 groups showed statistic significance(P<0.05). Curative effects comparison of stenocardia: the total effective rate of treatment group was 87.8%, the total effective rate of control group was 82.9%. The comparison of stenocardia between the two groups showed no statistic significance(P>0.05). Curative effects comparisonthat presented by electrocardiogram: The total effective rate of treatment group was 75.6%, the total effective rate of control group was 61.0%, the comparison of electrocardiogram between the two groups showed no statistic significance.(P>0.05). The VEGF level of both group showed significantly increase after treatment(P<0.05), in which the VEGF level of treatment group was markedly higher than that of control group. The ET-1, hs-CRP, IL-1β levels of both groups exhibited significant drop after treatment(P<0.05), in which the decrease of ET-1, hs-CRP, IL-1β level of treatment group was better than that of control group(P<0.05).(2) The alcohol extractions of both Fufangteng Yixin decoction and Yixin decoction were better than water extractions. The toxicity text on rats of alcohol extraction proofed its safety. Medicated rats’ serum prepared by twice human daily dose of Fufangteng Yixin decoction and 4 times human daily dose of Yixin decoction exhibited the highest HUMCE activity after H/R injuries.(3)Module control group: compared with Fufangtent Yixin decoction group, Yixin decoction group, CAP1 group, CAP2 group, and CAP3 group, the cell viability, NO content, NOS activity, VEGF concentration, VEGFR-2 concentration, and i NOS-m RNA genetic expression of module control group showed decrease(P<0.05). The caspase-3 positive cell number also decreased by 9.87%, 3.1%, 14.26%, 7.38%, and 4.17%(P<0.05). The ET-1, ECE protein, and ET-m RNA genetic expression exhibited significant increase(P<0.05). Fufangteng Yixin decoction group: compare with Yixin decoction group, the cell viability, NO content, NOS activity, VEGF concentration, VEGFR-2 concentration and i NOS-m RNA genetic expression of Yixin decoction group decreased(P<0.05), and the ET-1, ECE protein expression and ET-m RNA genetic expression of this group markedly increased(P<0.05). Compare with CAP2 group, the cell viability, NO content of CAP2 group decreased(P<0.05), its caspase-3 positive cell number increased, and its ET-1, ECE protein expression and theET-m RNA genetic expression markedly increased(P<0.05). Compared with CAP3 group, the cell viability, NO content, VEGF concentration, VEGFR-2 concentration of CAP3 group decreased(P<0.05),its caspase-3 positive cell number reduced by 5.70%(P<0.05), its ET-1, ECE protein expression and ET-m RNA genetic expression markedly increased(P<0.05). Yixin decoction group: compared with CAP1, CAP2, and CAP3 groups, the cell viability, NO content, NOS activity, VEGF concentration, and VEGFR-2 concentration of CAP1, CAP2, and CAP3 groups all increased(P<0.05), the ET-1, ECE protein and ET-m RNA genetic expression all reduced(P<0.05). The i NOS-m RNA genetic expression of CAP1 and CAP2 groups increased(P<0.05), the i NOS-m RNA genetic expression of CAP3 group reduced(P<0.05).Conclusions:(1)Fufangteng Yixin decoction is effective in relieving the angina of patients with thoracic obstruction induced by qi deficiency and blood stasis(Chronic stable angina), and improving the syndromes and ischemia. Otherwise, it can promote the excretion of VEGF but reduce the excretion of ET-1, hs-CRP and IL-1β, thus to maintain the balance of endogenous active peptides, inhibit the inflammatory response, protect the microvascular endothelial cells of endocardium and promote their functional recovery.(2)Both Fufangteng Yixin decoction and Yixin decotion are effective in increasing the proliferation activity of HUMCE after H/R injury; promoting the excretion of VEGF, VEGFR and NO, NOS; improving the expression of i NOS-m RNA and reducing the expression of ET-1, ECE and ET-m RNA. Besides, they also have the functions of maintaining the balance of ET/NO, stabilizing the completion of HUMCE after H/R injury and improving their functions. Fufangteng Yixin decoction is better than Yixin decoction and 10-4mol/L captopril in protecting the HUMCE after injury, but similar to 10-3mol/L captopril. |
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