基于心力衰竭大鼠PPARα和β3表达筛选鹿红方有效组分

Screening of Effective Components of Luhong Recipe Based on PPARα and β3 Expression in Rats with Heart Failure

目的:观察不同组分鹿红方对心力衰竭大鼠过氧化物酶增殖物激活受体(PPARα)和β3受体表达的影响,筛选鹿红方最佳有效组分。方法:分离鹿红方全方水煎液,分别用蒸馏水、30%乙醇、60%乙醇和95%乙醇洗脱,得到4种提取液,分别成为具有不同有效组分的A、B、C、D液。采用冠脉结扎法制备充血性心力衰竭大鼠模型,将96只SD大鼠随机分为16组,每组6只。1~15组均予造模,第16组为假手术组。1~14组,即A(1)、B(2)、AB(3)、C(4)、AC(5)、BC(6)、ABC(7)、D(8)、AD(9)、BD(10)、ABD(11)、CD(12)、ACD(13)、BCD(14)灌胃相应的鹿红方提取液,第15组灌胃曲美他嗪悬液,第16组灌胃0.9%Na Cl溶液,均5 ml/d,连续干预8周后取样。运用q PCR法检测各组大鼠心肌组织中PPARα表达,运用IHC法检测心肌组织β3表达的变化。结果:各治疗组(第1~15组)与假手术组(第16组)依次比较,第13组分的PPARα表达更加接近假手术组,差异无统计学意义(P>0.05);鹿红方各组分(第1~14组)与曲美他嗪组(第15组)依次比较,鹿红方各组分PPARα相对表达量相对更少,差异有统计学意义(P<0.05)。各治疗组(第1~15组)与假手术组(第16组)依次比较,鹿红方第4、第13组分的β3受体表达更加接近假手术组,差异无统计学意义(P>0.05);鹿红方各组分(第1~14组)与曲美他嗪组(第15组)依次比较,鹿红方第2、4、6、10、11、12、13、14组分的β3受体表达均高于曲美他嗪组,差异有统计学意义(P<0.05)。结论:筛选出鹿红方最佳有效组分为第13组分(ACD),即蒸馏水、60%乙醇和95%乙醇洗脱提取液的组合。

Objective: To observe the effects of different components of Luhong Recipe on the expressions of peroxisome proliferator activated receptor α(PPARα) and β3 receptor in rats with heart failure,and screen the best effective components of Luhong Recipe. Methods: The decoction of Luhong Recipe was separated by elution with distilled water,30% ethanol,60%ethanol and 95% ethanol respectively,and 4 kinds of extract with different effective components were obtained and labeled as A,B,C and D solution respectively. 96 SD rats were randomly divided into 16 groups,6 rats in each group. Except the No. 16 group as the sham operation group,rat model of congestive heart failure was established by coronary artery ligation in the other groups. From No. 1 to No. 14 group in turn,the rats were treated with A solution,B solution,A and B mixed solutions,C solution,A and C mixed solutions,B and C mixed solutions,A,B and C mixed solutions,D solution,A and D mixed solutions,B and D mixed solutions,A,B and D mixed solutions,C and D mixed solutions,A,C and D mixed solutions and B,C and D mixed solutions for intragastric administration,respectively. And the rats in the No. 15 group and No. 16 group were treated with trimetazidine suspension and normal saline respectively. The dosage of administration in all groups was 5 ml everyday. After intervention for continuous 8 weeks,the samples were collected. The expression of PPARα in myocardial tissue of rats in each group was detected by q PCR,and the expression of β3 in myocardial tissue was detected by IHC. Results: For the comparison between the sham operation group(No. 16 group) and the treatment groups(No. 1-15 groups) in turn,there was no statistically significant difference on PPARα expression between the sham operation group(No. 16 group) and No. 13 group(treated with A,C and D mixed solutions)(P > 0. 05). For the comparison between the trimetazidine group(No. 15 group) and Luhong Recipe extracts groups(No. 1-14 groups) in turn,the relative expression of PPARα in different components extracts of Luhong Recipe was lower,with statistical difference(P < 0. 05). For the comparison between the sham operation group(No. 16 group) and the treatment groups(No. 1-15 groups) in turn,there was no statistically significant difference on β3 receptor expression between the sham operation group(No. 16 group) and No. 4 group(treated with C solution) or No. 13 group(treated with A,C and D mixed solutions)(P > 0. 05). For the comparison between the trimetazidine group(No. 15 group) and Luhong Recipe extracts groups(No. 1-14 groups) in turn,the β3 receptor expression in No. 2 group(treated with B solution),No. 4 group(treated with C solution),No. 6 group(treated with B and C mixed solutions),No. 10 group(treated with B and D mixed solutions),No. 11group(treated with A,B and D mixed solutions),No. 12 group(treated with C and D mixed solutions),No. 13 group(treated with A,C and D mixed solutions) and No. 14 group(treated with B,C and D mixed solutions) was all higher than that in the trimetazidine group(No. 15 group),with statistical difference(P < 0. 05). Conclusion: After screening,the best effective component of Luhong Recipe is No. 13 mixed extract,that is the mixture of elution extracts of distilled water,60% ethanol and95% ethanol.