Preparation Of Cidan Injection And Its Effect On The Isolated Heart Pump Function Of Rat

Objective:To determine the Cidan injection on the basis of standard preparation technique and quality control, this essay is to determine the effect of Cidan injection on the normal and hypoxia/reperfusion injury in isolated rat heart pump function.Method:1. The preparation of Cidan injectionAccording to preparation process reference of Salvia miltiorrhiza injection and Acanthopanax injection of the Ministry of Health of The People’s Republic of China requirement, Cidan injection was prepared.2. To establish the quality control standards of Cidan injectionAccording to the chemical composition of Cidan injection materials and process of Cidan injection, the main chemical components by high performance liquid chromatography of Cidan injection were analyzed, on the basis of qualitative and quantitative analysis methods, the quality control standard of Cidan injection is established.3. Effect of Cidan injection on cardiac pump function in isolated normal rat heart.SD rats were randomly divided into 4 groups, including control group, Acanthopanax injection group, Salvia miltiorrhiza injection group and Cidan injection group,8 rats in each group. Groups were given normal saline 10 mL·L-1, Acanthopanax injection 10 mL·L-1, Salvia miltiorrhiza injection 10 mL·L-1, Cidan injection 8 mL·L-1 perfusion solution. Record the index parameters of before and during medicines perfusion for 5 minutes. Calculate all index parameters which increased, and observe the effect of Cidan injection on isolated cardiac pump function in rats.4. The dose-effect relationship research about effect of Cidan injection on isolated heart pump function in ratsSD rats were randomly divided into 5 dose groups for 1,2,4,8,16 mL·L-1 perfusion nutrient, respectively,5 rats in each group. Record the index parameters of before and during medicines perfusion for 1 and 5 minutes. To calculate every index parameters increasing, and observe the dose-effect relationship of Cidan injection on isolated cardiac pump function in rats.5. The effect of Cidan injection on hypoxia/reperfusion in isolated rat cardiac pump functionSD rats were randomly divided into 5 groups, including control group, model group, Acanthopanax injection group, Salvia miltiorrhiza injection group and Cidan injection group,8 rats in each group. Drug groups were given Acanthopanax injection 10 mL·L-1, Salvia miltiorrhiza injection 10 mL·L-1, Cidan injection 8 mL·L-1. Control group was perfused normal saline 10 mL·L-1 perfusion solution. The control group were normal continues infusion for 110 min. After 20 min of stabilization, hearts from model group were stopped perfusion for 30 min to induce hypoxia followed by reperfusion for 60 min, and the drug groups were done as aforementioned group. Observe the effect of Cidan injection on hypoxia/reperfusion in isolated rat cardiac pump function.Results:1. The preparation of Cidan injection The experimental results showed that, Acanthopanax senticosus Harms 750 g, Salvia miltiorrhiza 600 g is from Acanthopanax senticosus (calculated as syringoside):Salvia miltiorrhiza (calculated as protocatechualdehyde) 1.25:1. For Cidan injection preparation, water extraction and alcohol precipitation method were used. Two times ethanol precipitation of Cidan water extracts, followed by water precipitation, add 0.1% activated carbon, boiling, filtration, and sterilization.2. The establishment of the quality control standards of Cidan injectionThe experimental results presents the 1mL of product was contain more than 8.95 mg of Danshensu, protocatechuic aldehyde was greater than 2:79 mg, rosmarinic acid was over 2.38 mg, salvianolic acid B was more than 17.18 mg and syringoside was more than 0.49 mg.3. Effect of Cidan injection on cardiac pump function in isolated normal rat heart.The results showed that, after 5 min of perfusion, Acanthopanax injection, Salvia miltiorrhiza injection and Cidan injection elicited reduce HR by extremely significant difference in comparison with the previous administration (p<0.01); moreover, compared between Cidan injection with Ciwujia injection and Danshen injection, there was extremely significant difference (p<0.01). The Cidan injection with Ciwujia injection and Danshen injection groups were extremely statistical significantly increase in LVSP, LVDP,+dp/dtmax in comparison with before administration (P<0.01) and compared between groups (P<0.05, P<0.01). Moreover, all of three groups were able to decreased LVEDP and - dp/dtmax, there were extremely significant difference in comparison with before administration (P<0.01); while compare between Cidan injection and Ciwujia injection was no significant difference (P>0.05); whereas the comparison with Danshen injection was extremely significant difference (P<0.01,p<0.05).4. Dose-response relationship of Cidan injection on isolated heart pump function in rat.The present data demonstrate that Cidan injection (dose of 1 mL·L-1 to 8 mL·L-1) elicited increase cardiac pump function by dose-manner, and presents the obvious correlation between groups. However, greater than 16 mL·L-1 of dosing, the effects began to decline.5. The effect of Cidan injection on hypoxia/reperfusion in isolated rat cardiac pump functionThe recovery rate of heart rate in medicine group decreased by very significant difference in comparison with the model group (P<0.01). On the other hand, Cidan injection compared with Ciwujia injection shown no significantly different (P>0.05) but was a significant difference in comparison with Danshen injection (P<0.01). The three groups were able to increase the recovery rate of LVDP,+dp/dtmax and - dp/dtmax by extremely significant difference in comparison with the model group (P<0.01) and between groups (P<0.01).In addition, after reperfusion for 60 min, the activity of LDH and CK in the model group were significantly higher than control group (P<0.01). The CK and LDH activities of the medicine perfusion group were very significantly decreased when compared with the model group (P<0.01). Moreover, Cidan injection compared with Ciwujia injection and Danshen injection shown extremely significant difference (P<0.05, P<0.01, respectively).Conclusions:1. Cidan injection was significantly increase heart pump function in rat, and the effect was higher than that of Salvia miltiorrhiza injection and Acanthopanax senticosus injection.2. Effect of Cidan injection on cardiac pump function increasing in normal rats in dose of 1 mL·L-1 to 8 mL·L-1, demonstrate that linear relationship between groups.3. Cidan injection has significantly improve cardiac pump function in hypoxia/reperfusion isolated rat heart, and the improving effect were higher than that of Salvia miltiorrhiza injection and Acanthopanax injection.4. Cidan injection has more protective effect than Salvia miltiorrhiza injection and Acanthopanax injection on myocardial injury induced by hypoxia/reperfusion.