TGP Interfere In Primary Rat Myocardial Cells With Infection Of Coxsackie Virus B₃ By Method Of Serum Pharmacology

Objective: To investigate the effects of total glucosides of paeony for primary rat myocardial cells with infection of Coxsackie virus B3 by method of serum pharmacology and provide the theoretical basis for the clinical therapy of viral myocarditis. Methods: The study is divided into three parts. The first one is cultured primary rat myocardial cells. 15 newborn Sprague-Dawley rats of 1-3 days age were prepared. The ventricles muscle were take out to make into myocardial cells with no germ after they were digested with 0.25 per cent of trypsinase. The myocardial cells were moved into the culture bottle to culture them. The second part is preparation of each groups of medicine serum. Tirty 6-8 week SD big rats is divided into 6 groups , five male rats in each groups. They are low doses of TGP group(50mg/kg.d), middle doses of TGP group(100mg/kg.d), high doses of TGP group(200mg/kg.d), Shuanghuanglian group(60mg/kg.d), interferon group and normal serum group. Each rat of interferon group were injected with 5×105U interferon one time per day, the normal serum group doesn't give any medicine. Remaining groups were intragastric administration according to propotional dose of each group two times per day. All groups were give medicine for three days. After end time of giving medicine we adopts 10 ml blood from each rat's heart to make into six groups of the medicine serum. The toxicity of the medicine serum were measured with double deliquation method after their interfering myocardial cell. According to the CPE the eventual density of medicine serum is 20%. The third part is that medicine serum interferes in primary rat myocardial cells with infection of Coxsackie virus B3. The rat myocardial cell with forming single layer were divided into 8 groups:①cell matched control group,②virus matched control group,③low doses of TGP group,④middle doses of TGP group,⑤high doses of TGP group,⑥interferon group,⑦Shuanghuanglian group,⑧normal serum group. Maintenance liquid with the 100TCID50 CVB3 were added in each hole of single layer cell except the cell matched control group. At the same time each group were added in matched medicine serum 0.1 ml/hole. we cultivate them in 37℃5% CO2 incubator and stop cultivation when CPE of the virus matched control group appear ++++ effect. Take out each group of cell supernatant to metry cTnI, CK, LDH with the ELISA method, play all groups of cell down the light mirror to observate variety of cell appearance and measure cell livability by the MTT method. Results: (1) The cell livability of high doses of TGP group, interferon group and Shuanghuanglian group is 90±9 and 85±10 and 85±9 percent in turn. The virus matched control cell livability is 43±6 percent. Comparing their mean we draw a conclusion of P<0.05. While the cell livability of low doses of TGP group, middle doses of TGP group and normal serum group is 45±3, 48±6, 42±7 percent in turn, it shows no difference in comparing with the virus matched control group(P>0.05). After the myocardial cells were Interfered for 24 hours the LDH, CK, cTnI in the cell supernatant of high doses of TGP group, Shuanghuanglian group and interferon group are obviously lower than the virus matched control group(P<0.05). But when low doses of TGP group, middle doses of TGP group and normal serum group were compared with the virus matched control group, it shows no difference (P>0.05). Conclusion: The TGP group with high dose relieved the cytopathogenic effect, decreased cTnI, CK, LDH of the cell supernatant and has protective action on the primary rat myocardial cells with infection of CVB3.