Mechanism of Zhishi Xiebai Guizhitang in Treating AS Based on Regulation of Cholesterol Metabolism in Foam Cells by TRPA1 / 中国实验方剂学杂志

ObjectiveTo explore the effect and mechanism of Zhishi Xiebai Guizhitang on the progression of atherosclerosis （AS） mice based on the regulation of cholesterol metabolism in foam cells by transient receptor potential channel ankyrin 1 （TRPA1）. MethodThe AS model was established on apolipoprotein E knockout （ApoE-/-） mice with a high-fat diet. The mice were randomly divided into low-dose， middle-dose， and high-dose groups of Zhishi Xiebai Guizhitang （2.97， 5.94， 11.88 g·kg-1） and simvastatin group （0.002 g·kg-1）， and the drug was administered along with a high-fat diet. C57BL/6J mice were fed an ordinary diet as a normal group. After the above process， the aorta and serum of mice were taken. The pathological changes of the aortic root were observed by hematoxylin-eosin （HE） staining. The lipid plaques in the aorta were observed by gross oil redness. Serum levels of total cholesterol （TC）， triglyceride （TG）， low density lipoprotein cholesterol （LDL-C）， and high density lipoprotein cholesterol （HDL-C） were detected， and the levels of interleukin-1β （IL-1β） and interleukin-18 （IL-18） were detected by enzyme-linked immunosorbent assay （ELISA）. Western blot and immunohistochemical method were used to analyze the expression of TRPA1， ATP-binding cassette transporter A1 （ABCA1）， ATP-binding cassette transporter G1 （ABCG1）， and mannose receptor （CD206）. ResultFrom the perspective of drug efficacy， compared with the normal group， pathological changes such as plaque， a large number of foam cells， and cholesterol crystals appeared in the aorta of the model group， and the serum levels of TC， LDL-C， IL-1β， and IL-18 were significantly increased （P<0.01）. The HDL-C level was significantly decreased （P<0.01）， and the CD206 level in aortic tissue was significantly decreased （P<0.01）. Compared with the model group， the lipid deposition in the aorta was alleviated in all drug administration groups. In addition， except for the high-dose group of Zhishi Xiebai Guizhitang， all drug administration groups could significantly decrease the levels of TC and LDL-C （P<0.01）. In terms of inflammation， except for the middle-dose group of Zhishi Xiebai Guizhitang， the levels of IL-1β and IL-18 were significantly decreased in all drug administration groups （P<0.05）. Moreover， Zhishi Xiebai Guizhitang could also up-regulate the levels of CD206， and the difference was significant in the middle-dose and high-dose groups （P<0.05）. From the perspective of mechanism， the expression levels of TRPA1， ABCA1， and ABCG1 in the aorta in the model group were lower than those in the normal group （P<0.05）. Compared with the model group， all drug administration groups significantly increased the expression of TRPA1 in the aorta （P<0.05）， and the expressions of ABCA1 and ABCG1 were increased. The differences in the middle-dose and high-dose groups and the simvastatin group were significant （P<0.05）， which was basically consistent with the trend of immunohistochemical results. ConclusionZhishi Xiebai Guizhitang can effectively reduce blood lipid and inflammation levels and inhibit the formation of aortic plaque. The mechanism may be explained as follows： the expressions of ABCA1 and ABCG1 downstream are increased through TRPA1， which promotes cholesterol outflow in foam cells， thereby regulating cholesterol metabolism， intervening in inflammation level to a certain extent， and finally treating AS.

Modern Pharmacological Effect of Zhishi Xiebai Guizhitang： A Review / 中国实验方剂学杂志

Zhishi Xiebai Guizhitang is a classical prescription for the treatment of chest impediment with the method of warming Yang. It is included in the Catalogue of Ancient Classical Prescriptions issued by the National Administration of Traditional Chinese Medicine （First Batch）， with the effect of activating Yang， dissipating mass， moving Qi and resolving phlegm. Its main symptoms include chest fullness and pain， or even chest pain radiating to the back， wheezing， coughing， shortness of breath， and Qi reversal from the hypochondrium. In modern traditional Chinese medicine， Zhishi Xiebai Guizhitang is clinically used in the treatment of cardiovascular system， digestive system， respiratory system and other diseases， among which coronary heart disease， unstable angina pectoris， myocardial infarction， sinus bradycardia and other cardiovascular diseases have particularly significant effects. This paper reviewed the pharmacological studies of Zhishi Xiebai Guizhitang in the past 10 years. The results showed that each single medicine and the whole prescription alleviated the above cardiovascular diseases to a certain extent， with the pharmacological effects of improving intravascular environment， myocardial ischemia， myocardial ischemia-reperfusion injury， and myocardial hypoxia， anti-inflammation， plaque stabilisation， etc.， and the pharmacological mechanism involved the regulation of relevant active substances in vivo as well as related signaling pathways and ion channels， mainly including thromboxane B2 （TXB2）， prostacyclin I2（PGI2） and phosphatidylinositol 3-kinases/protein kinase B/endothelial nitric oxide synthase （PI3K/Akt/eNOS） signaling pathways， and ATP-sensitive potassium channels. In addition， the relationship between the pharmacological effects of some single medicines and transient receptor potential ankyrin 1 （TRPA1） has been reported that TRPA1 is a key to understanding the mechanism of Zhishi Xiebai Guizhitang in treating cardiovascular diseases， which is worth of further study.

Mechanism of Zhishi Xiebai Guizhitang in Alleviating Myocardial Injury in Rats with Myocardial Infarction Based on TNF/NF-κB Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo investigate the effect of Zhishi Xiebai Guizhitang （ZXGT） on isoproterenol （ISO）-induced myocardial infarction （MI） in rats through the tumor necrosis factor/nuclear factor-κB （TNF/NF-κB） signaling pathway. MethodForty-eight SD rats were randomly divided into control group （blank）， model group， perindopril group （4 mg·kg-1）， ZXGT group （24.4 g·kg-1）， ZXGT +inhibitor group （ZXGT， 24.4 g·kg-1， TNF-α receptor inhibitor R7050， 5 mg·kg-1）， and an inhibitor group （R7050， 5 mg·kg-1）， with eight rats in each group. The rats in each group were orally administered with their respective drugs for 7 days. Additionally， in the ZXGT + inhibitor group and the inhibitor group， R7050 was injected intraperitoneally at a dose of 5 mg·kg-1 on the 6th and 7th days. Except for the control group， all other groups were given intraperitoneal injections of ISO for 2 consecutive days to induce MI in rats. On the 7th day of the experiment， the rats were anesthetized 30 min after ISO injection， and their electrocardiograms （ECGs） were recorded to observe ST-segment elevation. Small animal echocardiography was used to measure global longitudinal strain （GLS） and cardiac synchrony. Blood samples were collected from the abdominal aorta to measure the levels of serum cardiac troponin T （cTnT）， creatine kinase-MB （CK-MB）， lactate dehydrogenase （LDH）， tumor necrosis factor-α （TNF-α）， and interleukin-1β （IL-1β）. Histopathological changes in myocardial tissue were observed using hematoxylin-eosin （HE） staining. Immunohistochemistry （IHC） was used to detect the expression of TNF-α， NF-κB p65， and p-NF-κB p65 proteins in myocardial tissue. Western blot was performed to measure the expression of tumor necrosis factor receptor 1 （TNFR1）， tumor necrosis factor receptor-associated factor 2 （TRAF2）， transforming growth factor-beta-activated kinase 1 （TAK1）， NF-κB inhibitory protein alpha （IκBα）， phosphorylated （p）-IκBα， NF-κB p65， and p-NF-κB p65 proteins in myocardial tissue. ResultCompared with the control group， the model group showed significant ST segment elevation on the ECG （P<0.01）， increased GLS， and reduced cardiac synchrony on echocardiography （P<0.01）. Histopathological examination revealed extensive myocardial necrosis. Furthermore， the serum levels of cTnT， CK-MB， LDH， TNF-α， and IL-1β were significantly increased （P<0.01）， and the expression levels of TNF-α， TNFR1， TRAF2， TAK1， p-IκBα， and p-NF-κB p65 proteins in myocardial tissue were significantly elevated （P<0.01）， while the expression level of IκBα was significantly decreased （P<0.01）. Compared with the model group， the perindopril group， the ZXGT group， the ZXGT + inhibitor group， and the inhibitor group rats showed a significant reduction in ST-segment elevation on the ECG （P<0.05， P<0.01）， improvement in GLS and cardiac synchrony （P<0.05， P<0.01）， a decrease in the area of myocardial necrosis， and reduced serum levels of cTnT， CK-MB， LDH， TNF-α， and IL-1β （P<0.01）. Additionally， the ZXGT group， the ZXGT + inhibitor group， and the inhibitor group downregulated the increased TNF-α， TNFR1， TRAF2， TAK1， p-IκBα， and p-NF-κB p65 protein expression levels and upregulated IκBα expression levels in the myocardial tissue （P<0.05， P<0.01）. No significant differences were observed between the ZXGT group and the ZXGT + inhibitor group or the inhibitor group. ConclusionZXGT can protect against ISO-induced myocardial injury in rats and improve cardiac function， and its mechanism of action may be related to the regulation of the TNF/NF-κB signaling pathway.

Interpretation of Theory of Weiluo Tongxin and Its Potential Molecular Mechanism Based on Zhishi Xiebai Guizhitang / 中国实验方剂学杂志

The scientific interpretation of the connotation of traditional Chinese medicine （TCM） theory is an important part of the development of TCM. Combined ancient classic theories with modern science and technology is a new path for the innovative development of TCM theory. Based on this， taking Zhishi Xiebai Guizhitang as an example， the molecular mining technology of integrative pharmacology-based research platform of TCM V2.0 （TCMIP V2.0） was used to analyze the mechanism of Zhishi Xiebai Guizhitang in preventing and treating gastropathic stomachache. A total of 220 chemical components in Zhishi Xiebai Guizhitang were obtained， and 674 targets were involved， of which 12 core targets directly affected angina pectoris and gastroesophageal reflux disease， including insulin （INS）， glyceraldehyde-3-phosphate dehydrogenase （GAPDH）， RAC-α serine/threonine-protein kinase 1 （AKT1）， tumour protein p53 （TP53）， albumin （Alb）， mitogen-activated protein kinase 3 （MAPK3）， interleukin-6 （IL-6）， etc. And tumor necrosis factor （TNF） signaling pathway， phosphatidylinositol 3-kinase/protein kinase B （PI3K/Akt） signaling pathway， IL-17 signaling pathway， hypoxia-inducible factor-1 （HIF-1） signaling pathway and other 121 pathways involved in these related diseases. All of these are the biological basis of Weiluo Tongxin theory. Combing the classical theories of TCM combined with the exploring the molecular mechanism of representative prescriptions can provide a demonstration and reference for the scientific connotation research of TCM theory.