A Study Into Delayed Protective Function Of The Precondition Of Jiawei Danshen Decoction To The Rat's Myocardial Cell And Its Cell Signal Transduction Mechanism

Purpose: To observe the delayed protective function of the precondition of Jiawei Danshen decoction(JDD) to the rat's myocardial cell and its cell signal transduction mechanism.Method: The experiment contains three steps.Step 1:Rat that were born 72 hours ago were chosen and rat myocardial cell, which had been cultured for 72 hours, was divided at random into 6 groups. The normal group was cultured in a usual way; the serum contrast group with 50% rat serum; and the medicine-serum group with 50% medicine serum containing JDD. The Hypoxic/reoxygenation(H/R) group was made to have three-hour's hypoxia and then reoxygenated for an hour. The Hypoxic preconditioning(HPC) group and the JDD preconditioning(JDDPC) group were given HPC and JDD precondition separately. After 24 hours, both of them were made to have three-hour's hypoxia and then reoxygenated for an hour. At the end of this step, the cell survival rate of each group was identified by TB straining while LDH and CK by colorimetry.Step 2:The rat myocardial cell, cultured for 72 hours, was divided into 8 groups at random. The normal group, the serum contrast group, the medicine-serum contrast group, the H/R group, the HPC group and the JDDPC group were treated the same as in Step 1. The HPC+PMB group was added PMB before precondition, and its following process was the same as the HPC group. The JDDPC + PMB group was added PMB before precondition, and the following process was the same as the JDDPC group . At the end of this step, the cell survival rate of each group was identified by TB straining, LDH and CK by colormetry, and PKC by PepTag(?) non-radioactive protein kinase reagent.Step 3:The rat myocardial cell, cultural for 72 hours, was divided into 8 groups at random. The normal group, the serum contrast group, the medicine-serum contrast group, the H/R group, the HPC group and the JDDPC group were treated the same as in Step 1. The HPC+GLI group was added GLI before precondition, and its following process was the same as the HPC group. The JDDPC + GLI group was added GLI before precondition, and the following process was the same as the JDDPC group . At the end of this step, the cell survival rate of each group was identified by TB straining, LDH and CK by colormetry. The concentration of calcium ion inside the cell was tested by Fura-2/AM, as the indicator, and the HSP 70m RNA by RT-PCR.Result: The cell survival rate of HPC group and JDDPC group is obviously higher than the H/R group (P<0.01), while LDH and CK lower than H/R group (P<0.01). The cell survival rate, LDH and CK of HPC+PMB group and HPC+GLI group are similar to the H/R group (P>0.05). The cell survival rate of JDDPC + PMB group and JDD PC+ GLI group are higher than the H/R group (P<0.01), but lower than HPC and the JDDPC group (P<0.01). The LDH and CK of them are lower than the H/R group (P<0.01), but higher than HPC and JDDPC group.The PKC activity and HSP70mRNA of the HPC group and JDDPC group are obviously higher than H/R group (P<0.01), but the concentration of calcium ion inside the cell is obviously lower than the H/R group (P<0.01). The PKC activity of HPC+PMB group and the HSP70mRNA and concentration of calcium ion inside the cell of HPC+GLI group are similar to H/R group (P>0.05). PKC activity of the JDDPC + PMB group and HSP 70mRNA of JDDPC + GLI group are higher than H/R group (P<0.01 or P<0.05), but lower than HPC and the JDDPC group. The concentration of calcium ion inside the cell of JDD PC+ GLI group is lower than the H/R group (P<0.01), but higher than HPC and the JDDPC group (P<0.01).Conclusion: The precondition of JDD is of delayed protective function. PMB, the inhibitor of PKC, and GLI, of K_ATP, can completely stop the delayed protective function of HPC, which means PKC is the signal transduction pathway of HPC and K_ATP is the final effector, K_ATP can reduce calcium overloan and induce a high convey of HSP70. PMB, the inhibitor of PKC, and GLI, of K_ATP, can not completely stop the delayed protective function of the precondition of JDD, which means the precondition of JDD does not completely play its delayed protective function through PKC signal transduction pathway and K_ATP pathway, indicating that there are many pathways of the signal transduction in JDDPC.