| Purpose1.Clinical research:Randomized,single blind,parallel control method was used to observe the clinical efficacy of Guanxinping in the treatment of elderly stable coronary artery disease(SCAD)with carotid artery plaque(CAP),and the mechanism of its anti-inflammatory reaction was also investigated;2.Animal experiment:The intervention effect of Guanxinping on inflammatory reaction of atherosclerosis was studied,and the anti-inflammatory mechanism of Guanxinping was discussed from the perspective of Klotho/FGF21/FGFR1 axis by establish atherosclerosis,ApoE-/-mice models.Method1.Clinical research:from March 2020 to September 2021,60 patients with elderly stable coronary artery disease(SCAD)and carotid atherosclerosis diagnosed and treated in Affiliated Hospital of Nanjing University of Chinese Medicine were selected and included according to the standard.They were divided into the control group(n=30)and the treatment group(n=30)by using random,single blind,parallel control method.Both groups were administered with basis treatment.The control group was treated with Yixinshu tablets,and the treatment group was treated with Guanxinping tablets.The efficacy of both groups was evaluated after 3 months.We evaluated symptom score of traditional Chinese and western symptom score,C-IMT and Crouse score of carotid ultrasound,comprehensive therapeutic effect of diseases,comprehensive therapeutic effect of traditional Chinese medicine,serum Klotho,FGF21 levels,and serum inflammatory factor TNF-α,IL-6,CRP levels,and the expression level of FGFR1 protein extracted from peripheral blood monocytes after treatment.2.Animal experiment30 ApoE-/-male mice were randomly divided into 5 groups(blank group-C group,model group-M group,low dose group of being administrated with Guanxinping-L group,high dose group of being administrated with Guanxinping-H group and atorvastatin group-A group).Except that the blank group was fed with the ordinary diet,the other four groups were fed with high-fat diet to establish the AS model.After the model was constructed,the four groups of mice with high-fat diet were administrated with different drugs(model group-normal saline,low dose group of Guanxinping-0.74g/kg,high dose group of Guanxinping-1.47g/kg and atorvastatin group-4.11mg/kg)for 12 weeks by means of oral treatment.HE was used to observe the pathological changes of aorta and heart tissues,Elisa,qPCR,WB and other techniques were used to detect the level of serum inflammatory factors and the expression of Klotho/FGF21/FGFR1 axis related proteins in myocardial cells.Result1.Clinical efficacy and anti-aging factor levels(1)Angina pectoris symptom score and ECG efficacy evaluationThe result of angina pectoris symptom score and ECG efficacy evaluation after treatmentCompare the Angina pectoris symptom score and ECG efficacy evaluation among the earlier and the after of the treatment,with the P-value is less than 0.01,there is a significant improvement both two groups,and with the P-value is less than 0.05 the treatment group creates distinctly healing effect.After the treatment,the treatment group holds better TCM symptom scores than the same variable of the control group,and the P-vale of the TCM symptoms benefits is less than 0.05.(2)Judgment of comprehensive clinical efficacyThe result of comprehensive clinical efficacy:Compared with the total effective rate,the treatment group holds a meaningful mathematics leadership from the same variable of the control group.Under the P-value is less than 0.05,the mathematics leadership is large enough for the statistically significant.(3)TCM symptom score and TCM symptom comprehensive curative effect judgmentThe result of TCM symptom score and TCM symptom comprehensive curative effect judgment after treatment.Compare the score and comprehensive curative effect among the earlier and the after of the treatment,with the P-value is less than 0.01,there is meaningful mathematics variation between two groups which is large enough for the statistically significant.After the treatment,the treatment group holds better TCM symptom scores than the same variable of the control group,and the P-vale of the TCM symptoms benefits is less than 0.05.(4)Carotid artery ultrasound C-IMT,Crouse scoreThe result of Carotid artery ultrasound C-IMT and Crouse score after treatment.Even through the variation between the earlier and after of treatment is distinct in intra-group with the P-value is less than 0.05.On the statistically signification,with the P-value is larger than 0.05,there is a judgement that those two groups hold the similar/same scores.(5)serum TNF-α,IL-6 and CRP resultsOn the statistically signification,before treatment those two groups hold the similar serum inflammatory factors.And in the same situation,there exists a huge scope of decline of inflammatory factors after treatment with the P-value less than 0.001.The treatment group holds mathematics lower levels of serum inflammatory factors than the variables from the control group.And on the statistically signification,the variation is meaningful with the P-value is less than 0.05.(6)Klotho,FGF21,FGFR1 resultsThe result of Klotho,FGF21 after treatmentThere exists a distinctly elevation in intra-group about the serum Klotho concentration with the P-value is less than 0.01.And in the same situation the treatment group holds a distinctly elevation of the serum Klotho concentration upon the control group with the P-value is less than 0.05.There exists a distinctly variation of the both groups about the serum FGF21 concentration with the P-value is less than 0.01.And on the statistically signification,with the P-value is less than 0.05,there is a judgement that those two groups hold the similar/same serum FGF21 concentration.Before treatment,the level of serum FGFR1 in the treatment group was similar to that in the control group,but there was no statistical significance between the two groups(P>0.05).After treatment,the level of serum FGFR1 in the treatment group was significantly decreased,with significant statistical difference(P<0.01).The level of serum FGFR1 in the treatment group was lower than that in the control group,with statistical significance(P<0.05).2.Animal experiment(1)HE staining results of aortaHE staining of the aorta showed that the intima of the blood vessel wall in the blank group was clear without plaque;The intima of the model group was obviously thickened,and plaque formation was visible;In the treatment group,the intima of the blood vessel wall was slightly thickened,and the plaque was significantly smaller;The low-dose Guanxinping group had more severe atherosclerosis than the high-dose Guanxinping group.(2)HE staining results of heartHE staining of heart tissue showed that in the model group,the tissue around the coronary artery was significantly increased,inflammatory cells infiltration was found,and myocardial fibers were disintegrated,fragmented or lost.The myocardial structure in the treatment group was significantly improved.Compared with the other treatment groups,the tissue around the coronary artery was increased in the low-dose Guanxinping group,and there was infiltration of inflammatory cells.(3)Serum inflammatory factor levelsCompared with blank group,the level of inflammatory factors in model group was significantly increased,with significant statistical difference(P<0.001);Compared with the model group,the serum levels of inflammatory factors CRP,IL-6 and TNF-α in the high-dose Chinese medicine group were significantly lower(P<0.001).Meanwhile,the inhibitory effect of the high-dose Chinese medicine group on the serum inflammatory factors was better than that of the atorvastatin group,and the difference was statistically significant(P<0.01).(4)Serum Klotho,FGF21,FGFR1 factor levelsKlotho level in model group was significantly lower than that in blank group,and the difference was statistically significant(P<0.001).The levels of FGF21 and FGFR1 were significantly increased(P<0.001).Compared with model group,serum Klotho level in atorvastatin group and TCM high-dose group was significantly increased(P<0.001),indicating that atorvastatin and coronary heart can increase serum Klotho level on average.In the atorvastatin group,the level of FGF21 was significantly decreased with significant difference(P<0.001),but the level of FGF21 was increased in the TCM group,which may be the cause of the compensatory increase of FGF21,and the specific mechanism needs further study.The serum level of FGFR1 decreased significantly in both atorvastatin and TCM high-dose groups(P<0.001).(5)Protein expression of Klotho,FGF21,FGFR1 in heart tissueIn the heart tissue of mice in each group,the expression levels of Klotho,FGF21 and FGFR1 proteins in the model group were the lowest,and there was a statistical difference from other groups with the P-value is larger than 0.05.On the statistically signification,with the P-value is larger than 0.05,there is a judgement that the Model group and the low-dose group hold the similar levels of the Caspase-3 proteins expression.But in the same situation the high-dose Guanxinping group and the Atorvastatin group holds distinctly elevations than those two groups.(6)mRNA expression of Klotho,FGF21 and FGFR1 factors in heart tissueComparison of Klotho mRNA:Compared with the blank group,the expression of Klotho gene in the other four groups was down-regulated to varying degrees,and the down-regulation in the model group was the most obvious(P<0.01).Compared with the model group,the gene expression of the three treatment groups was all up-regulated.Among them,the up-regulation in the Atorvastatin group was the most obvious(P<0.05),followed by the high-dose traditional Chinese medicine group(P<0.05).Comparison of FGF21 mRNA:On the statistically signification,with the P-value is less than 0.01,the model group holds a distinctly descent curve about the FGF21 gene expression with the blank group.Besides the gene expression in the observation groups was down-regulated to varying degrees.There were statistical differences between the Atorvastatin group,the high-dose traditional Chinese medicine group and the model group(P<0.05).Comparison of FGFR1 mRNA:On the statistically signification,with the P-value is less than 0.01,there exists an ascending curve of the gene expression of FGFR1 in ApoE-/-mice on a high-fat diet towards the blank group.But in the similar situation,the three observation groups hold a descent curve of gene expression after the drug intervention and a distinctly variation with the model group.And with specifically noted that the descent curve of Atorvastatin group has the steepest slope.(7)Protein expression of Caspase3、ERK1/2、P65 in heart tissueCaspase3 protein level:On the statistically signification,with the P-value is less than 0.01,the model group holds distinctly elevations of the level of the Caspase-3 proteins expression towards the blank group.And for the same variable both the atorvastatin group and guanxinping high-dose group hold the similar-shape descent curve.Besides there is a judgement that those two groups hold the similar counts.And with the P-value is larger than 0.05 and P-value is less than 0.01,the low-dose Chinese medicine group holds an indistinctly variation with the model group.Erk1/2 protein level:On the statistically signification,with the P-value is less than 0.05,the model group holds distinctly elevations of the level of erKl/2 protein towards the blank group.The Western medicine group and the Chinese medicine high-dose group holds a distinctly descent curve of the level of erKl/2 protein but without distinctly variation towards the model group.P65 protein level:On the statistically signification,with the P-value is less than 0.01,the model group holds an ascending curve of the level of the P65 protein towards the blank group but the treatment groups hold opposite.With the same situation there exists diversities among the atorvastatin group,guanxinping high-dose group and the model group,but opposite in the three treatment groups.Conclusion1.It is found that Guanxinping can obviously improve the clinical symptoms,TCM syndromes and ECG scores of elderly patients with stable coronary artery disease complicated and carotid plaque,reduce the carotid intima-media thickness,reduce the plaque thickness,increase the Klotho level,and decrease the levels of serum inflammatory factors,FGF21 and FGFR1.So,just to prove the point,it has obvious anti-inflammatory effect.2.It is found that Guanxinping can obviously improve atherosclerosis of ApoE-/atherosclerosis model mice,improve myocardial fibrosis in mice,the mechanism is suspected to be related to the regulation of Klotho/FGF21/FGFR1 axis inhibition of inflammatory response. |
PDF Full Text Request