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The Effect of Shenqi Injection on Tumor Necrosis Factor Receptor Associated Factor 6 in Mice with Viral Myocarditis

Author: WangChao
Tutor: ChenJinShui
School: Fujian Medical
Course: Integrative Medicine
Keywords: Viral myocarditis (VMC) Tumor necrosis factor receptor associated factor 6 Shenqi fuzheng injection
CLC: R285
Type: Master's thesis
Year: 2011
Downloads: 38
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Abstract


Objective: To investigate the pathogenesis of Tumor Necrosis Factor receptor associated factor 6(TRAF6) and the mechanism of Shenqi Fuzheng injection (Shenqi) in viral myocarditis (VMC), the thisis explored the effect of Shenqi on TRAF6 in mice with VMC induced by CVB3.Method: A total of 120 4-6-week-old Balb/c male mice were randomly divided into 5 groups:normal control group(group A), CVB3 group(group B,), ribavirin group ( group C ) , low-dose Shenqi Fuzheng injection(shortened as Shenqi)intervention group(group D), high-dose Shenqi intervention group(group E). Mice in the infected group were inoculated intraperitoneally with 0.1 ml of 1×102 TCID50 CVB3 diluted in Eagle’s minimal essential medium (EMEM) solution. Mice in group A were given intraperitoneal injection of EMEM solution without CVB3 at the same dosage. Mice in group A and group B were treated with i.p. normal saline solution 0.4ml; the others were given i.p.0.9mL saline containing2.7mg ribavirin (group C), 0.4ml Shenqi (groupD)and 0.9ml Shenqi (group E)respectively after inoculation 0.5 hours. Then, mice in each group were treated as ever on the daily basis for 30 days respectively.The death status was observed during the study. Mice were euthanized after taking the eyeball for blood and hearts were collected randomly on the third day, on the 10th day and on the 30th day. The body weight (BW) was detected. Myocardial pathology integration was caculated according to hematoxylin-eosin staining (HE). The change of the ultrastructure in myocardium was also observed with electron microscopy. The blood was homogenized to determination serum myocardial enzymes. Reverse Transcription Polymerase Chain Reaction (RT-PCR) were used to investigate the expression of TRAF6 mRNA. Streptavidin-peroxidase coiugated (SP) and Western-blot technique were also used to investigate the expression of TRAF6 protein in mice myocardial tissue at different times.Result: 1. As the BW of normal control group increased normally, the BW of CVB3 group were gained slowly on the third day after inoculation, decreased obviously on the tenth day and increased on thirtieth day, but it was still significantly lower than that of the control group(P<0.01). 2. Compared with normal control group, the histopathologic score of myocardium in CVB3 group increased dramatically on the third day after inoculation, and reached a peak value on the tenth day (P<0.01)and was reduced on the thirtieth day; the similar changes were observed in the serum myocardial enzymes,the expression levels of TRAF6mRNA and protein. On 3th,10th and 30th day after infection, Levels of TRAF6mRNA and protein in myocardial tissue of CVB3 groups were positively correlated with the degree of myocardial histopathologic score[r=0.977(mRNA), 0.961(SP), 0.960(Western-blot), P<0.01]. When myocardial ultrastructure were observed by electron microscopy it was indicated that the myocardial fibers were disorderly arranged, myofibrils were broken, lysis and even vanished partially, mitochondria were swelling and degenerated, T-tubule and sarcoplasmic reticulum (SR) were dilated. The damage of myofibrils and mitochondria were lightened greatly, but interstitial fibrosis increased obviously on the thirtieth day. 3. Compared with CVB3 group, Shenqi could lower the total mortality, myocardial histopathologic score and the serum myocardial enzymes; decreased the damage of myocardial ultrastructure and inhibited the expression of TRAF6mRNA and protein in myocardial tissue in a dosage-dependent manner. There was a significant difference between the low-dose Shenqi intervention group and the CVB3 group(P<0.05~0.01).Conclusion:1. CVB3 can induce the myocardial damage in histopathology and increase the serum myocardial enzymes level. 2. Shenqi can decrease the damage of myocardial histopathology in VMC mice induced by CVB3, reduce the serum myocardial enzymes level and lower the mortality. 3. The expression of TRAF6mRNA and protein were increased in myocardial tissues of mice with VMC induced by CVB3. 4. Shenqi can inhibit the expression of TRAF6mRNA and protein in myocardial tissues of VMC mice induced by CVB3. 5. Shenqi can improve the survival rate and lighten the histopathologic damage of myocardium through down-regulating the expression of TRAF6.

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