Abstract:Aim To study the effect of astragaloside on mRNA expression and protein production of inflammatory cytokines,including TNF-α,IL-1β,IL-6 and IL-10,in Viral Myocarditis in Mice with CVB3. Methods Fifty Balb/c mice were randomized into five groups (n=10):normal control group,given 0.1 mL of EMEM by intraperitoneal injection,were teated with saline 0.1 mL with gavage after 30 minites of injection for 1 week; viral myocarditis control group,inoculated intraperitoneally with 0.1 mL of 1×102 TCID50 CVB3 diluted in Eagles minimal essential medium (EMEM) solution were given saline 0.1 ml with gavage after 30 minites of injection for 1 week; astragaloside low-dose intervention group,middle-dose intervention group and high-dose intervention group,inoculated intraperitoneally with 0.1 mL of 1×102 TCID50 CVB3 diluted in EMEM were treated with 1%,3%,9% astragaloside [0.07,0.2 and 0.6 mg/(kg·d)],respectively) 0.1 mL solution after 30 minites of injection,respectively with gavage for 1 week. After fourteen days,the mice were sacrificed and their hearts were taken and cut into two equal parts:one was used to determine mRNA expression of cardiac cytokines by RT-PCR and another was used to measure protein production of cytokines by Western blot. Results No expression of inflammatory cytokines mentioned above was found in normal control group. In viral myocarditis control group,both of these cytokines markedly increased (P<0. 01). Compared with viral myocarditis control group,the high-dose astragaloside group displayed significant reduction of mRNA expression and protein production of TNF-α,IL-1β and IL-6,and marked increase of IL-10 mRNA expression and protein production (P<0. 01). Conclusion Astragaloside markedly reduced pro inflammatory cytokines,and increased inflammation preventing cytokine. The mechanism needs to be elucidated.

Abstract:Aim To study protective effect and the anti-oxidize mechanism of Astragaloside on viral myocarditis in mice with CVB3. Methods Fifty Balb/c mice were randomized into five groups(n=10): normal control group,given 0.1 mL of EMEM by intraperitoneal injection,were teated with saline 0.1 mL with gavage after 30 minities of injection for 1 week;viral myocarditis Control group,inoculated intraperitoneally with 0.1 mL of 1×102 TCID50 CVB3 diluted in Eagles minimal essential medium(EMEM) solution were given saline 0.1 mL with gavage after 30 minities of injection for 1 week;Astragaloside lowdose intervention group,middle-dose intervention group and high-dose intervention group,inoculated intraperitoneally with 0.1 mL of 1×102 TCID50 CVB3 diluted in Eagles minimal essential medium(EMEM) were treated with 1%,3% and 9% astragaloside 0.07,0.2 and 0.6 g/(kg·d),respectively] 0.1 mL solution after 30 minetes of injection,respectively with gavage for 1 week.Survival number,score of pathological changes,GSH-PX,CAT,SOD and CuZn-SOD-mRNA of myocardium were detected. Results The survival number was significantly improved in Balb/c mice treated with high dose astragaloside group than that in viral myocarditis control group(P<0.01).Heart function was better in high-dose intervention group than in the viral myocarditis control group,the activity of GSH-PX,CAT,SOD and CuZnSOD -mRNA levels was enhanced and heart function was improved in group high dose astragaloside group than in viral myocarditis control group(P<0.01). Conclusion The results showed that astragaloside can provide protection against viral myocarditis.This protective effect of astragaloside may be related to the maintenance of the antioxidant status of the heart in improving myocardial antioxidant enzymes activity .

Abstract:Aim To elucidate the effect of adrenocoticoids on viral myocarditis. Methods Two hundreds male 4-week-old Balb/c mice were divided into five groups randomly, including normal control group, infected control group, group of infected mice with adrenocorticoids treatment at early stage, group of infected mice with adrenocorticoids treatment at middle stage, group of infected mice with adrenocorticoids treatment at late stage. Normal control group mice were inoculated intraperitoneally with Eagle’s medium 0.1 mL. Infected group were inoculated intraperitoneally of 10 7.5 TCID50 Coxsackievirus B3 at a dose of 0.1 mL per mouse. Five mice of each group were sacrificed on 3, 7, 10, 14, 21, 30, 40 and 50 days post-inoculation. Pathologic examination and ultrastructure of heart, virus titer in heart by plaque-forming assay, serum cardiac troponin I with chemiluminescence immunoassay were examined. Results The myocardial histopathological score of mice in each infected group on day 7 to 10 after infection were significantly higher than that in normal control group, but no significant difference in each infected group. On day 14 after infection, the myocardial histopathological score of mice with adrenocorticoids treatment at middle stage were significantly lower than that in other infected groups, and no myocardial lesions were noted 30 days later post-infection while cardiac lesions could be noted in other infected groups. Virus titer in group of infected mice with adrenocorticoids treatment at early stage was significantly higher than that of in other infected group. Cardiac troponin I values were significantly greater in infected group than that in normal control group. There was a positive correlation between the level of cardiac troponin I and the change of myocardial histopathological score. Cardiac troponin I values was much lower after adrenocorticoids therapy, especially in group of infected mice with adrenocorticoids treatment at middle stage. Conclusions Adrenocorticoids therapy ameliorated the severity of myocardial lesions in viral myocarditis, especially used in middle stage of the disease. Adrenocorticoids therapy reduced the inflammatory infiltration in myocardium, inhibited autoimmune response. Adrenocorticoids therapy reduced the level of serum cardiac troponin I in viral myocarditis.

Abstract:Aim To measurement the inhibitory effect of Momordicin on the activity of caspase 3 and apoptosis of myocardium in BALB/C mice with coxsackievirus B3 myocarditis. Method Momordicin was purificated from Momordica charantia L by our previous reported methods. Four group animals were used in this experiment, such as disease group (DG), normal group (NG), Momordicin group (MG), and Momordicin therapy group (MTG). Five animals were killed at the 3rd,the 7th,and the 14th days in each group, and all were killed at the 21st day. According to the method of Calbiochem Company, the activity of caspase 3 from the heart of BALB/C mice was measured, and apoptosis was measured by terminal transferase mediated DNA nick end labeling assay (TUNEL, the method provided by Oncogene Company). Results ①The activity of caspase 3 were measurable at the 7th day in the DG (0.63±0.21 pmol/min, n=5). The activity of caspase 3 at the 14th day (10.9±1.5 pmol/min, n=5) and the 21st day (12.6±1.3 pmol/min, n=5) are higer than the 7th day, and the activity of the 21st was higher than 14th day (p<0.05). ②Only a animal has the activity of caspase 3 at the 21st day (0.41 pmol/min) in the HTG, no caspase 3 activity were detected in the animals of MG and NG. ③Only a few apoptotic cells were found at the 7th day in the DG, and more apoptotic cells were found among cardiac muscles at 14th and 21th days. Single apoptotic cardiomyocytes were also found at the outside of the pathological change areas, and no apoptotic cells were found in the animals of NG, MG, and MTG. Conclusions Distinct apoptosis were found in CVB3 viral myocarditis; the activitys of caspase 3 and apoptosis were found at the 7th day, and higher after the 7th day, single apoptotic cardiomyocytes were also found. The high dose Momordicin has distinct inhibitory effect on the activity of caspase 3 and apoptosis.