Study On The Medication Rule Of Patients With Pyrogenic Coronary Heart Disease And The Mechanism Of Huanglian Jiedu Decoction Based On Autophagy To Interfere With AS

ObjectiveCoronary heart disease is a common cl ini cal disease,and the incidence rate continues to rise in China.Modern research shows that noxious heat with blood stasis has become one of the basic pathogenesis of coronary heart disease.The climate in Lingnan is hot and humid,people eat more sweet taste,and it is easier to form noxious heat with blood stasis coronary heart disease.The purpose of this study was to investigate the rule of medication used in the treatment of pat ients with noxious heat with blood stasis type coronary heart disease in the First Affiliated Hospital of Guangzhou University of Chinese Medicine,and to provide evidence for clinical treatment.MethodsCollection of prescriptions for the treatment of coronary heart disease with noxious heat and blood stasis in the Department of Cardiology,the First Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine from January 2018 to January 2019.Application of "Traditional Chinese Medicine Aided Platform" software for data collation and mining,analysis and summary of hospi tal medications for patients with noxious heat with blood stasis type coronary heart disease.Results1.238 cases were included,238 prescriptions,and the drug totaled 185 flavors.2.There are 19 herbs with frequency ≥30,and 13 drugs with frequency≥60,namely Danshen(216),safflower(198),Chuanxiong(197),Chishao(190),Mao Dongqing(176).),Astragalus(175),Fragrance(173),Astragalus(85),Jujube(83),Zhuru(69),Pinellia(67),Melon(65),and Corydalis(64).3.Set the support degree to 160 and the confidence level to 0.90.According to the frequency of drug combination,the order is from high to low,and 40 common drugs and combinations are obtained.The first three are Chuanxiong-Salvia,Honghua-Chuanxiong and Chishao.-Chuanxiong.4.Combine the experience judgment and the pre-reading of the data extracted by different parameters,set the correlation degree to 6,the penalty degree is 3,apply the hierarchical clustering analysis of complex system entropy clustering,and evolve 8 sets of core drug combinations.The first two combinations are "Mao Dongqing,Peach Kernel,Salvia Miltiorrhiza","Huang Wei,Mao Dongqing,Chishao,and Fragrance",and excavated 4 new prescriptions such as "Chuanxiong,Salvia,Safflower,and Scorpion".ConclusionIn our hospital,the treatment of noxious heat with blood stasis type coronary heart disease is more common with Qingrehuoxue traditional Chinese medicine.With the addition and subtraction of Yiqi and Huayu products,it reflects the clinical application of heat-toxic coronary heart disease to"clear heat and promote blood circulation,with spleen and qi,The treatment concept of "heating and phlegm".ObjectiveQingrejiedu method is a treatment for heat-toxic coronary heart disease and basic treatment.Exploring the atherosclerosis(AS)detoxification therapy in combination with the early stage of the team and the analysis of the medication rules of patients with heat-toxic coronary heart disease in our hospital.We selected Qingrejiedu Mingfang lluanglian Jiedu Decoction as a therapeutic drug,observed the effect of Huanglian Jiedu Decoction on AS in New Zealand white rabbits,and explored its mechanism of action from the perspective of macrophages autophagy.Methods1.Qualitative and quantitative detection of Iluanglian Jiedu Decoction lyophilized powder by liquid-mass spectrometry.2.The AS model of New Zealand white rabbit was constructed.HE staining and ELISA were used to observe the therapeutic effect of Huanglian Jiedu Decoction on rabbit AS and the effect of AS-related serum inflammation.3.The effects of Huanglian Jiedu Decoction on the autophagy phenotype of New Zealand white rabbits were observed by immunohistochemistry and Western blot.4.The macrophage cell line raw line 264.7 was cultured and induced to become foam cells by oxidized low density lipoprotein(ox-LDL).Oil red 0 staining was used to verify the induction of foam cells.5.RTCA method was used to screen the concentration range of Huanglian Jiedu Decoction lyophilized powder solution(HL),and the cells were treated with HL at different time and concentration to observe the effect on autophagy phenotype.The intervention time and concentration of HL were selected.6.Cell electron microscopy,immunofluorescence staining,PCR and Western blot were used to verify the expression of autophagy in normal,model and HL intervention groups.7.Western blot was used to detect the expression of P-PI3K/PI3K,P-Akt/Akt and P-mTOR/mTOR,and the HL group was treated with PI3K agonist 740Y-P,AKt agonist SC79 and mTOR agonist MHY,respectively.The expression levels of the pathway proteins P-PI3K/PI3K,P-Akt/Akt,P-mTOR/mTOR and autophagy-related proteins LC3B and P62 were detected.Results1.Qualitative detection of Huanglian Jiedu Decoction lyophilized powder containing 27 components such as palmatine,eucommia alcohol,rhein and berberine;quantitative detection of seven components in the content of berberine 0.55 mg/g,coptisine 0.72 mg/g,Geniposide 5.02mg/g,Phellodendron 0.46 mg/g,Epirubicin 2.98 mg/g,Palmatine 1.92 mg/g,Baicalin 0.87mg/g.2.In HE staining,plaque formation was observed in the model group compared with the normal group.The plaques in the low,middle and high dose groups of Huanglian Jiedu Decoction and the atorvastatin group in the positive drug group were reduced compared with the model group,and the Huanglian Jiedu Decoction was medium and high.The plaque reduction was significant in the dose group and the atorvastatin group.3.Compared with the normal group,the expression levels of serum AS-associated inflammatory factors TNF-α and IL-1β in rabbit model group were significantly increased(P<0.05),IL-10 levels were decreased(P<0.05),and Huanglian Jiedu Decoction group was compared with the model.Compared with TNF-α and IL-1β,the expression levels of TNF-α and IL-1β were significantly decreased(P<0.05),and IL-10 levels were increased(P<0.05).4.Immunohistochemistry and Western blot analysis showed that the expression levels of autophagy-related proteins LC3B and bclin-1 in the model group were significantly lower than those in the normal group(P<0.05).The blood group of Huanglian Jiedu Decoction was compared with the model group.The expression level of bclin-1 was increased(P<0.05).5.Cell oil red 0 staining showed that the cholesterol emission of the model group was significantly higher than that of the normal group(P<0.05),and the Huanglian Jiedu Decoction group was significantly lower than the model group(P<0.05).6.The RTCA real-time test showed that the HL concentration was 48.83 μ g/ml and below,and the cell proliferation was not affected.The 40 μ g/ml intervention foam cells showed more autophagy at different time points than the model group,and the autophagy level was the highest at 2 hours.The foam cells were treated with 10 μ g/ml,20 μ g/ml,40 μ g/ml Huanglian Jiedu Decoction lyophilized powder for 2 hours.The autophagy levels of the three intervention groups were higher than those of the model group,and the autophagy level was the highest at 40 μ g/ml.7.Real-time quantitative PCR and Western blot showed that the mRNA and protein levels of PI3K,Akt and mTOR in the model group were significantly higher than those in the normal group(P<0.05).The mRNA and protein of PI3K,Akt and mTOR in the HL intervention group compared with the model group.The expression level of the expression decreased(P<0.05).The expression of P-PI3K/PI3K,P-Akt/Akt and P-mfOR/mTOR in the model group was significantly higher than that in the normal group(P<0.05).The expression levels of P-PI3K/PI3K,P-Akt/Akt and P-mTOR/mTOR were lower than those in the model.group(P<0.05).The expression of LC313 was compared between 740Y-P group,SC79 group and MIIY group compared with HI,group.The expression of P62 was significantly increased(P<0.01),and the expression of P62 was significantly increased i n 740Y-P group compared with HL group(P<0.01).The expression of P-PI3K/PI3K was significantly increased in SC79 group and HL group.The expression of P-Akt/Akt was increased(P<0.01);the expression of P-mTOR/mTOR was increased in the MIIY group compared with the HL group(P<0.01).Conclusion Huanglian jiedu Decoction can improve atherosclerosis by inhibiting the mechanism of autophagy by inhibiting PI3K-Akt-mTOR pathway.