The Correlation Of The Relative Expression Of Myeloperoxidase And Praziquantel Concentration On Blood And The Effect Of Chinese Herbal Medicine In Carassius Auratus

As a leukocyte enzyme, myeloperoxidase（MPO） is widely present in animals, and is involved in various physiological responses in vivo. When the body is stimulated and the stress reaction is produced, myeloperoxidase content increased rapidly, showing strong oxidation activity, killing of invading microorganisms and enhancing immunity.The strong oxidizing property of myeloperoxidase pushs forward a immense influence on the oxidative metabolism of drugs. Praziquantel is a quinoline pyrazine derivatives,and widely used in aquaculture as an anti parasite drug. In this experiment, the praziquantel used as model drug, crucian carp used as a model organism, analysis of the relationship between myeloperoxidase m RNA expression and the concentration of praziquantel of the crucian carp after single dose oral administration of praziquantel,and we studied the effects of three kinds of Chinese herbal medicine（astragalus,bupleurum chinensis and coptis chinensis） on praziquantel pharmacokinetics and myeloperoxidase m RNA expression. Try to establish a method for the evaluation of fish in vivo drug residues with myeloperoxidase genetic.The main research and results in this paper are as follows:1.The correlation of the relative expression of myeloperoxidase（MPO） and praziquantel（PZQ） concentration on blood of crucian carp, Carassius auratusTo research the association of the relative expression of myeloperoxidase（MPO）m RNA and praziquantel（PZQ） concentration on the blood of crucian carp, Carassius auratus, after oral administration of 10 mg/kg PZQ, the relative expression of MPO and plasma concentration were explored using the real-time fluorescent quantitative PCR（RT-PCR） with the reference gene of β-actin and high performance liquid chromatography（HPLC）. The results showed that PZQ could be quickly absorbed into the blood and eliminated rapidly. And concentration and time data of PZQ in blood could be best described by a two-compartmental open pharmacokinetic model. The peak concentration of PZQ was 2.85 μg/m L in the first hour and the concentration of PZQ was belowed detection limit after 96 hours. What’s more, the peak value in the relative expression of MPO m RNA also appeared in the first hour, in the 0.25 h, 0.5 h, 1 h, 3 h and 6 h the relative expression of myeloperoxidase（MPO） m RNA were etremely singnificant difference with the control group（P < 0.01）, in the 12 h and 48 h the dufference was significant（P<0.05）,then gradually decreased to the relatively low value that was no significant difference with the control groupuntil 48 hours. In addition,the difference between the control group and the blank control group was 0.993, which showed that the full price feed and the mouth irrigation method had no effect on the experimental results. Correlation analysis indicated that the correlation coefficient of the MPO m RNA relative expression amount and PZQ concentration in blood was highly correlated（r=0.96）. That suggested MPO may participate in the oxidative metabolism of PZQ.2.Effect of Astragalus, Bupleurum chinensis and Coptis chinensis on praziquantel pharmacokinetics of crucian carp, Carassius auratusThe carp was firstly continuous fed the feed added astragalus, bupleurum chinensis and coptis chinensis for 30 days, and in the control group, fed the feed without the Chinese herbal. Then, the praziquantel concentrations of blood samples collected at 0.25 h, 0.5 h, 1 h, 3 h, 6 h, 12 h, 24 h, 48 h and 96 h after oral administration of 10 mg/kg PZQ were simultaneously detected using the high performance liquid chromatography.Pharmacokinetic analysis was performed using the DAS 3.0 program. The results showed that the pharmacokinetic parameters were gained based on the classical compartmental model analysis; Praziquantel concentrations-time profiles all were best described by a two-compartmental open pharmacokinetic model, and praziquantel concentrations of blood showed the trend of drop after the rising. Just like the control group, the T_max values of praziquantel in carp fed astragalus, bupleurum chinensis also were 1h. But the T_max values in carp fed coptis chinensis was 3h. The C_max values in carp fed astragalus, bupleurum chinensis and coptis chinensis respectively were 2.50 mg/L,2.90 mg/L, 3.01 mg/L. While, the C_max values in the control group was 2.71 mg/L. The t1/2β of praziquantel in carp fed astragalus, bupleurum chinensis and coptis chinensis respectively were 78.69 h, 61.708 h and 56.967 h, which all were above the value of the control group（15.007 h）. The AUC（0-∞） in carp fed astragalus, bupleurum chinensis and coptis chinensis were136.591 mg/L·h, 75.009 mg/L·h, 77.69 mg/L·h, and 4.43, 2.44, and2.52 fold of the value in the control group, respectively. The CL/F value in carp fed astragalus, bupleurum chinensis and coptis chinensis were 0.073 L/kg, 0.133 L/kg,0.129 L/kg, respectively, which all were lower than the control group;Astragalus,bupleurum chinensis and coptis chinensis can improve the bioavailability of praziquantel, but they also greatly slow their metabolism and elimination, resulting in longer residue time. So, the using of herbs with praziquantel should be cautious in aquaculture.3.Effect of Astragalus, Bupleurum chinensis and Coptis chinensis on myeloperoxidase expression of crucian carp, Carassius auratusThe carp was firstly continuous fed the feed added astragalus, bupleurum chinensis and coptis chinensis for 30 days, and in the control group, fed the feed without the Chinese herbal. Then, the blood samples collected at 1 h after oral administration of10 mg/kg PZQ, in the blank control group, fed with basic diet and no oral administration of praziquantel. The relative expression of MPO was explored using the real-time fluorescent quantitative PCR（RT-PCR） with the reference gene of β-actin. The myeloperoxidase relative expression of blank control group to the control group was increased significantly, indicating that praziquantel induced the expression of myeloperoxidase in crucian carp, the myeloperoxidase relative expression of three kinds of Chinese herbal medicine group compared with the control group were decreased,indicating that the three kinds of Chinese herbal medicine on myeloperoxidase expression were inhibited.