1.Proteomic Analysis of Danlou Tablet in Improving Platelet Function for Treating Coronary Heart Disease with Phlegm-stasis Intermingling Syndrome in Minipigs
Ziyan WANG ; Ying LI ; Aoao WANG ; Hongxu MENG ; Yue SHI ; Yanlei MA ; Guoyuan ZHANG ; Lei LI ; Jianxun LIU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(5):41-53
ObjectiveThis paper aims to observe the role of Danlou tablet in treating coronary heart disease (CHD) with phlegm-stasis intermingling syndrome in minipigs by improving platelet function and explore the potential pharmacological mechanism of Danlou tablet in regulating platelet function by using proteomics technology. MethodsThirty Bama minipigs were randomly divided into a normal control group (6 pigs) and a high-fat diet group (24 pigs). After 2 weeks of high-fat diet feeding, the high-fat diet group was randomly subdivided into a model group, an atorvastatin group (1 mg·kg-1), and Danlou tablet groups (0.6 g·kg-1 and 0.3 g·kg-1). All groups continued to receive a high-fat diet for 8 weeks after the procedure. The normal control group was given a regular diet, underwent only coronary angiography, and did not receive an interventional injury procedure. The model group and each administration group were fed a high-fat diet. Two weeks later, they underwent a coronary angiography injury procedure. After the procedure, drugs were mixed into the feed every morning for 8 consecutive weeks, with the minipigs maintained on a continuous high-fat diet during this period. Quantitative proteomics technology was further used to study platelet proteins, and differential proteins were obtained by screening. Bioinformatics analysis was performed to analyze key regulatory proteins and biological pathways involved in the therapeutic effect of Danlou tablet on CHD with phlegm-stasis intermingling syndrome. ResultsCompared with the normal control group, the model group showed a significant increase in total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) of minipigs' serum (P<0.01), a significant shortening in prothrombin time of (PT) (P<0.01), a coagulation function index, and an increase in whole blood viscosity (P<0.01) and platelet aggregation rate (P<0.01). Moreover, the platelet morphology was altered, and the contents of endothelin-1 (ET-1) and nitric oxide (NO) were significantly increased (P<0.01). Hemodynamic parameters were obviously abnormal, including significantly decreased systolic blood pressure (SBP), diastolic blood pressure (DBP), mean arterial pressure (MAP), left ventricular systolic pressure (LVSP), and left ventricular maximal positive dp/dt (LV+dp/dtmax) (P<0.01). Left ventricular maximal negative dp/dt (LV-dp/dtmax) was significantly increased (P<0.01). Besides, there were myocardial cell hypertrophy, obvious edematous degeneration, massive interstitial inflammatory cell infiltration, high degree of fibrosis, and coronary endothelial atherosclerosis. TC and TG levels in minipigs' serum were significantly reduced in Danlou tablet groups with 0.6 g·kg-1 and 0.3 g·kg-1 (P<0.05, P<0.01), compared with those in the model group. LDL-C was decreased in the Danlou tablet group with 0.6 g·kg-1 (P<0.05). The whole blood viscosity under low and high shear conditions was significantly reduced in the Danlou tablet group with 0.6 g·kg-1 (P<0.05). In groups with all doses of Danlou tablet, maximum aggregation rate (MAR) and average aggregation rate (AAR) were significantly decreased (P<0.05, P<0.01), and platelets' morphological changes such as pseudopodia extension were reduced. ET-1 levels in the serum were significantly reduced. In the Danlou tablet group with 0.6 g·kg-1, NO level in the serum was reduced (P<0.05). In groups with all doses of Danlou tablet, DBP and MAP were significantly increased (P<0.05). In the Danlou tablet group with 0.6 g·kg-1, LVSP and LV+dp/dtmax were significantly increased (P<0.05, P<0.01), and LV-dp/dtmax was significantly decreased (P<0.05). In groups with all doses of Danlou tablet, edematous degeneration in myocardial tissue was milder, and coronary artery lesion degree was significantly alleviated. Compared with the normal control group, there were 94 differentially expressed proteins in the model group, including 81 up-regulated and 13 down-regulated proteins. Compared with the model group, the Danlou tablet group with 0.6 g·kg-1 showed 174 differentially expressed proteins, including 100 up-regulated and 74 down-regulated proteins. A total of 30 proteins were reversed after Danlou tablet intervention. Bioinformatics analysis revealed that its pharmacological mechanism may exert anti-platelet activation, aggregation, and adhesion effects through biological pathways such as regulation of actin cytoskeleton, platelet activation pathway, Fcγ receptor-mediated phagocytosis, as well as proteins such as growth factor receptor-bound protein 2 (GRB2), Ras-related C3 botulinum toxin substrate 2 (RAC2), RAC1, and heat shock protein 90 alpha family class A member 1 (HSP90AA1). ConclusionDanlou tablet can effectively reduce platelet activation and aggregation, exerting a good therapeutic effect on CHD with phlegm-stasis intermingling syndrome in minipigs. Its pharmacological mechanism may involve regulating biological pathways such as actin cytoskeleton and platelet activation pathway, as well as proteins like GRB2, RAC2, RAC1, and HSP90AA1, thereby exerting a pharmacological effect in anti-platelet activation, aggregation, and adhesion.
2.Establishment and Evaluation of Rat Model of Myocardial Ischemia-reperfusion Injury with Phlegm and Blood Stasis Blocking Collaterals Syndrome Based on Metabolomics
Longxiao HU ; Jiabei GAO ; Weihao MA ; Jieming LU ; Yunxiao GAO ; Yue YUAN ; Qiuyan ZHANG ; Xiaoxiao CHEN ; Yali SHI ; Jianxun LIU ; Junguo REN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(12):41-51
ObjectiveTo explore the feasibility, evaluation methods and metabolic differences of high-fat diet(HFD) combined with myocardial ischemia-reperfusion injury(MIRI) to establish a rat model of myocardial ischemia-reperfusion with phlegm and blood stasis blocking collaterals syndrome(PBSBCS). MethodsThirty-two SD rats were randomly divided into the sham operation, HFD, MIRI, and MIRI+HFD groups. Rats in the sham operation and MIRI groups were fed a standard diet(regular chow), while the HFD and MIRI+HFD groups received a HFD for 10 weeks. Rats in the MIRI and MIRI+HFD groups underwent myocardial ischemia-reperfusion surgery, while the sham operation group underwent only thread placement without ligation. Cardiac function was assessed via small-animal echocardiography, including left ventricular ejection fraction(EF), left ventricular fractional shortening(FS), cardiac output(CO), and stroke volume(SV). Serum levels of creatine kinase(CK), CK-MB, triglyceride(TG), total cholesterol(TC), high-density lipoprotein cholesterol(HDL-C), low-density lipoprotein cholesterol(LDL-C), lactate dehydrogenase(LDH), endothelin-1(ET-1), endothelial nitric oxide synthase(eNOS), tumor necrosis factor-α(TNF-α), interleukin-18(IL-18), oxidized LDL(ox-LDL), and cardiac troponin T(cTnT) were measured by biochemical assays and enzyme-linked immunosorbent assay(ELISA). Myocardial histopathology was evaluated via hematoxylin-eosin(HE) staining, while myocardial infarction and no-reflow area were assessed using 2,3,5-triphenyltetrazolium chloride(TTC), Evans blue, and thioflavin staining. Changes in syndrome characteristics[body weight, tongue surface red-green-blue [RGB] values, and pulse amplitude] of PBSBCS were recorded. Serum differential metabolites were analyzed by ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS). ResultsCompared with the sham operation group, the HFD and MIRI+HFD groups showed significant increases in body weight(P<0.01), RGB values and pulse amplitude decreased in the HFD, MIRI and MIRI+HFD groups, TC, TG, LDL-C and ox-LDL levels increased in the HFD and MIRI+HFD groups, while HDL-C decreased. Blood perfusion peak time and myocardial no-reflow area increased, serum eNOS level decreased, and CK-MB, LDH, and cTnT activities increased in the HFD, MIRI and MIRI+HFD groups(P<0.05, P<0.01). Whole blood viscosity was increased in the HFD group at medium shear rate, and in the MIRI and MIRI+HFD groups at low, medium and high shear rates(P<0.05, P<0.01). Platelet aggregation rate increased in the MIRI and MIRI+HFD groups, accompanied by elevated ET-1, TNF-α, and IL-18 levels, reduced cardiac function indices, expanded myocardial no-reflow and infarction areas, and increased serum CK, CK-MB, LDH, and cTnT activities(P<0.05, P<0.01). Compared with the MIRI group, the HFD and MIRI+HFD groups showed significant increase in body weight, TC, TG, LDL-C and ox-LDL levels, and significant decrease in HDL-C content(P<0.01). The MIRI+HFD group showed decrease in RGB values and pulse amplitude, and an increase in whole blood viscosity, platelet aggregation, blood perfusion peak time, myocardial no-reflow and infarction areas, elevated ET-1, TNF-α and IL-18 levels, decreased eNOS content, EF and SV, increased serum CK, CK-MB and cTnT activities, and worsened myocardial pathology(P<0.05). Compared with the HFD group, the MIRI+HFD group showed similar aggravated trends(P<0.05, P<0.01). Metabolomics results showed that 34 potential biomarkers involving 13 common metabolic pathways were identified in the MIRI+HFD group compared with the sham operation group. ConclusionThe MIRI group resembles blood stasis syndrome in hemodynamics and myocardial injury, and the HFD group mirrors phlegm-turbidity syndrome in lipid profiles and tongue characteristics. While the MIRI+HFD group aligns with PBSBCS in comprehensive indices, effectively simulating clinical features of coronary heart disease(CHD), which can be used for the evaluation of the pathological mechanism and pharmacodynamics of CHD with PBSBCS.
3.Establishment and Evaluation of Rat Model of Myocardial Ischemia-reperfusion Injury with Phlegm and Blood Stasis Blocking Collaterals Syndrome Based on Metabolomics
Longxiao HU ; Jiabei GAO ; Weihao MA ; Jieming LU ; Yunxiao GAO ; Yue YUAN ; Qiuyan ZHANG ; Xiaoxiao CHEN ; Yali SHI ; Jianxun LIU ; Junguo REN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(12):41-51
ObjectiveTo explore the feasibility, evaluation methods and metabolic differences of high-fat diet(HFD) combined with myocardial ischemia-reperfusion injury(MIRI) to establish a rat model of myocardial ischemia-reperfusion with phlegm and blood stasis blocking collaterals syndrome(PBSBCS). MethodsThirty-two SD rats were randomly divided into the sham operation, HFD, MIRI, and MIRI+HFD groups. Rats in the sham operation and MIRI groups were fed a standard diet(regular chow), while the HFD and MIRI+HFD groups received a HFD for 10 weeks. Rats in the MIRI and MIRI+HFD groups underwent myocardial ischemia-reperfusion surgery, while the sham operation group underwent only thread placement without ligation. Cardiac function was assessed via small-animal echocardiography, including left ventricular ejection fraction(EF), left ventricular fractional shortening(FS), cardiac output(CO), and stroke volume(SV). Serum levels of creatine kinase(CK), CK-MB, triglyceride(TG), total cholesterol(TC), high-density lipoprotein cholesterol(HDL-C), low-density lipoprotein cholesterol(LDL-C), lactate dehydrogenase(LDH), endothelin-1(ET-1), endothelial nitric oxide synthase(eNOS), tumor necrosis factor-α(TNF-α), interleukin-18(IL-18), oxidized LDL(ox-LDL), and cardiac troponin T(cTnT) were measured by biochemical assays and enzyme-linked immunosorbent assay(ELISA). Myocardial histopathology was evaluated via hematoxylin-eosin(HE) staining, while myocardial infarction and no-reflow area were assessed using 2,3,5-triphenyltetrazolium chloride(TTC), Evans blue, and thioflavin staining. Changes in syndrome characteristics[body weight, tongue surface red-green-blue [RGB] values, and pulse amplitude] of PBSBCS were recorded. Serum differential metabolites were analyzed by ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS). ResultsCompared with the sham operation group, the HFD and MIRI+HFD groups showed significant increases in body weight(P<0.01), RGB values and pulse amplitude decreased in the HFD, MIRI and MIRI+HFD groups, TC, TG, LDL-C and ox-LDL levels increased in the HFD and MIRI+HFD groups, while HDL-C decreased. Blood perfusion peak time and myocardial no-reflow area increased, serum eNOS level decreased, and CK-MB, LDH, and cTnT activities increased in the HFD, MIRI and MIRI+HFD groups(P<0.05, P<0.01). Whole blood viscosity was increased in the HFD group at medium shear rate, and in the MIRI and MIRI+HFD groups at low, medium and high shear rates(P<0.05, P<0.01). Platelet aggregation rate increased in the MIRI and MIRI+HFD groups, accompanied by elevated ET-1, TNF-α, and IL-18 levels, reduced cardiac function indices, expanded myocardial no-reflow and infarction areas, and increased serum CK, CK-MB, LDH, and cTnT activities(P<0.05, P<0.01). Compared with the MIRI group, the HFD and MIRI+HFD groups showed significant increase in body weight, TC, TG, LDL-C and ox-LDL levels, and significant decrease in HDL-C content(P<0.01). The MIRI+HFD group showed decrease in RGB values and pulse amplitude, and an increase in whole blood viscosity, platelet aggregation, blood perfusion peak time, myocardial no-reflow and infarction areas, elevated ET-1, TNF-α and IL-18 levels, decreased eNOS content, EF and SV, increased serum CK, CK-MB and cTnT activities, and worsened myocardial pathology(P<0.05). Compared with the HFD group, the MIRI+HFD group showed similar aggravated trends(P<0.05, P<0.01). Metabolomics results showed that 34 potential biomarkers involving 13 common metabolic pathways were identified in the MIRI+HFD group compared with the sham operation group. ConclusionThe MIRI group resembles blood stasis syndrome in hemodynamics and myocardial injury, and the HFD group mirrors phlegm-turbidity syndrome in lipid profiles and tongue characteristics. While the MIRI+HFD group aligns with PBSBCS in comprehensive indices, effectively simulating clinical features of coronary heart disease(CHD), which can be used for the evaluation of the pathological mechanism and pharmacodynamics of CHD with PBSBCS.
4.Application of dual chamber round tissue expander in immediate breast reconstruction.
Jianxun MA ; Xi BU ; Bi LI
Journal of Peking University(Health Sciences) 2025;57(1):166-171
OBJECTIVE:
To explore the application value of dual chamber round tissue expander in immediate breast reconstruction.
METHODS:
Sixteen patients, who had been provided immediate tissue expander/implant two-stage breast reconstruction using dual chamber round tissue expander in our hospital from March 2022 to October 2023, were involved in this study, and the relevant information was analyzed retrospectively. The overall design of the expander is a round shape, consisting of two equally divided semi-circular chambers. The two expansion chambers are connected by a silicone pad below and are respectively connected to their own water injection tubes. Both chambers are designed to expand unidirectionally towards the surface. The expansion principle, insertion process, and type of expander selection were investigated. The expansion effect and incidence of complications were summarized. The aesthetic effect of reconstructed breasts was evaluated from three aspects after stage Ⅱ surgery: the position of infra mammary fold, the breast protrusion, and the breast volume.
RESULTS:
Among sixteen patients in this study, three patients were selected with the type of 400 mL expander and thirteen patients were given the type of 600 mL expander. The median time of tissue expansion was 4.0 (2.0, 5.0) months, with an average volume of expansion of (538.8±111.7) mL. The average expansion ratio of upper/lower chamber was 45.4%±8.4%. The position of the infra mammary fold needed not to be adjusted during the prosthesis exchange process. All the patients were applied anatomical prostheses, and the median volume of the prosthesis was 395 (345, 410) mL. One patient developed seroma during expansion period, who got improved after local aspiration. The average follow-up time was (9.0±3.6) months. 81.3% (13/16) of the patients achieved an aesthetic evaluation of "Good" in breast reconstruction, and 75.0% (12/16) of the patients got a grade Ⅰ or grade Ⅱ capsule contracture of the prosthesis.
CONCLUSION
The application of dual chamber round tissue expander could effectively dilate the lower pole of the breast, personalize the expansion ratio of the upper and lower poles of the breast, and avoid the displacement of the expander during the expansion period. Therefore, it could provide a good foundation for subsequent prosthesis exchange.
Humans
;
Tissue Expansion Devices
;
Female
;
Mammaplasty/instrumentation*
;
Tissue Expansion/instrumentation*
;
Retrospective Studies
;
Adult
;
Middle Aged
;
Breast Neoplasms/surgery*
;
Breast Implants
;
Mastectomy
5.Effect of Longmu Piyan Prescription on Oxidative Stress in Atopic Dermatitis Mice
Mengxi MA ; Siqi WU ; Qingying WANG ; Yaqin LI ; Jinhe WANG ; Na LANG ; Jianxun REN
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(5):88-95
ObjectiveThis study observes the intervention effect of Longmu Piyan prescription on oxidative stress in BALB/c mice with atopic dermatitis (AD) induced by 2,4-dinitrochlorobenzene (DNCB) and explores its mechanism. MethodThe AD model was established using the method of DNCB sensitization on the back skin of BALB/c mice. Forty male BALB/c mice were randomly divided into a blank group, a model group, a vitamin C control group (0.5×10-3 mg·kg-1), and a Longmu Piyan prescription group (26 g·kg-1). Except for the blank group, other groups were sensitized with different concentrations of DNCB on the back to induce AD, and the blank group was treated with matrix coating. The gastric administration was started on the seventh day after sensitization with 2% DNCB and on the 24th day after sensitization with 0.2% DNCB continuously for 21 days. The changes in skin lesions of each group were directly observed after the experiment. Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of interleukin (IL)-4, tumor necrosis factor (TNF)-α, immunoglobulin E (IgE), and reactive oxygen species (ROS) in the serum of each group. The total antioxidant capacity determination kit-trace method (ABTS method) was used to measure the level of total antioxidant capacity (TAOC) in serum. The Hematoxylin eosin (HE) staining method was used to observe the pathological and morphological changes of the skin lesion site. The immunohistochemical method was used to detect the expression of thymic stromal lymphopoietin (TSLP) in the skin lesion site. Western blot was used to detect the expression of filaggrin (FLG) in the dorsal skin lesions. ResultThe results showed that compared with the blank group, the skin lesion score of the model group mice was significantly increased (P<0.01), and HE staining showed characteristic pathological changes of AD in the skin lesion site. At the same time, the expression of TSLP in the skin lesion was significantly increased, and that of FLG was reduced (P<0.05). The levels of TNF-α, IL-4, IgE, and ROS in serum increased, while the activity of TAOC decreased (P<0.01). Compared with the model group, the Longmu Piyan prescription group showed a significant decrease in skin lesion scores and a significant improvement in skin lesion pathology. At the same time, the expression of TSLP decreased, and the expression of FLG increased in the skin lesions (P<0.05). In addition, compared with the model group, the serum levels of TNF-α, IL-4, IgE, and ROS also decreased to varying degrees (P<0.05,P<0.01), and TAOC activity increased in the Longmu Piyan prescription group (P<0.01). ConclusionThere is a significant correlation among the degree of oxidative stress, the severity of skin lesions in AD, and the levels of inflammatory cytokines. Longmu Piyandu prescription can improve AD-like skin lesions in BALB/c mice by promoting ROS clearance, enhancing TAOC, and inhibiting oxidative stress, thus protecting the skin barrier and reducing inflammation.
6.Procedure for correction of inverted nipple using tiny incision with primary breast ducts reserved
Youchen XIA ; Bi LI ; Jianxun MA ; Lijun ZOU
Chinese Journal of Medical Aesthetics and Cosmetology 2023;29(3):171-173
Objective:To discuss the procedure for correction of inverted nipple using tiny incision with primary breast ducts reserved.Methods:A total of 35 patients (63 sides) with primary inverted nipples from January 2006 to March 2019 were reviewed retrospectively. Tiny radial incisions were made on the areola around the base of the inverted nipple which had been pulled out. Without skin removed, shorten fiber bundles which caused nipple inverted were totally cut and released. While the primary breast ducts were preserved, purse-string suture was taken around the base of the nipple. The nipple protector was prepared by ourselves, and the nipple was pulled and suspended for 2-6 months.Results:Sixty-three sides of 35 patients with inverted nipples were successfully corrected by this minimally invasive surgery. There was no nipple necrosis. One patient developed mild swelling 3 weeks after operation, and the swelling subsided after symptomatic anti-inflammatory treatment. The average follow-up period was 39 months. After removing the nipple protector, 2 sides (2/63) had a certain degree of recurrence. The rest of the nipples had ideal shape, no obvious scar, good nipple feeling, and retained the possibility of lactation.Conclusions:The procedure for correction of inverted nipple using tiny incision with primary breast ducts reserved has advantages of minimal invasion, safety, less pain, while retaining the possibility of lactation in the future. The clinical effect is satisfactory. It is especially suitable for the correction of type Ⅰ and type Ⅱ inverted nipples.
7.Recent advance in predictive value of blood biomarkers in post-stroke cognitive impairment
Tingting YANG ; Yanru CHEN ; Jianxun CAO ; Lin HAN ; Yuxia MA
Chinese Journal of Neuromedicine 2023;22(3):318-324
Post-stroke cognitive impairment (PSCI), one of the important complications of stroke, seriously affects the quality of life of these patients. PSCI is an important cause of disease burden of stroke. In recent years, more and more evidences show that blood biomarkers are of great significance in PSCI diagnosis, and the detection of blood biomarkers is relatively simple and more suitable for clinical application. Therefore, this paper sorts out the values of 5 blood biomarkers, nerve injury marker, metabolic biomarker, inflammatory biomarker, oxidative stress marker and other biomarker, in diagnosing PSCI, to provide references for early diagnosis and intervention of PSCI.
8.Application of tailor-tack technology in vertical mastopexy
Xi BU ; Jianxun MA ; Yue LANG ; Bi LI
Chinese Journal of Medical Aesthetics and Cosmetology 2022;28(2):127-129
Objective:To explore the value of tailor-tack technology in vertical mastopexy and to evaluate the clinical effect of vertical mastopexy in correcting breast ptosis.Methods:From April 2010 to August 2020, 47 women aged 18 to 51 years took part in the study, and the tailor-tack technology was used to ensure the amount of redundant skin removal. The patients' degree of ptosis was moderate or severe, and the average age was 38.2 years.Results:All the cases had their drainage tube removed 2-3 days after surgery and the wound stitches removed 12-14 days after surgery. All patients had tight and full breasts after surgery. Patients were satisfied with their new breast shapes. After surgery, no patients showed early complications such as necrosis of nipple-areola complex or skin, poor wound healing or abnormal nipple sensation. Follow-up lasted for 6 months to 5 years, and there were no long-term complications such as recurrence of breast ptosis, poor nipple shape, scar flatten and enlargement.Conclusions:Tailor-Tack technology is useful in vertical mastopexy. It can be helpful to evaluate the amount of skin removal, effectively avoiding the situation of insufficient or excessive skin removal. Therefore, this is a desirable clinical skill in mastopexy.
9.Effect of Buyang Huanwutang on Platelet Function and Inflammatory Cytokine in Rat Model of Acute Blood Stasis Induced by Multiple Factors
Ziyan WANG ; Lei LI ; Jianxun LIU ; Hongxu MENG ; Yue SHI ; Yanlei MA ; Aoao WANG ; Yue YOU
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(21):1-9
ObjectiveTo explore the effect of Buyang Huanwutang (BYHWT) on platelet function and inflammatory cytokines in the rat model of acute blood stasis. MethodThe model of acute blood stasis was established with SD rats by ice water bath combined with injection of epinephrine. Rats were randomly assigned into four groups: normal group, model group, BYHWT (3.2 g·kg-1) group, and aspirin (60 mg·kg-1) group. The rats were injected with epinephrine hydrochloride on day 8 after 7 days of modeling. The macroscopic indexes of triditional Chinese medicine (TCM) syndrome including tongue manifestation and pulse manifestation were observed, while hemorheological indexes, blood coagulation, and platelet aggregation were detected. The serum levels of the inflammatory cytokine matrix metalloprotein-9 (MMP-9) and the adhesion factor intercellular adhesion molecule-1 (ICAM-1) and were determined by enzyme-linked immunosorbent assay (ELISA). ResultThe pulse distention of rats in the model group was lower than that in the normal group (P<0.01), while BYHWT improved the pulse distention of the rats with the syndrome of blood stasis (P<0.01). In the model group, the tongue showed the characteristics of blood stasis syndrome, with dark purple veins at the tongue bottom and lower values of R, G, B on the tongue surface than those in the normal group (P<0.01), which, however, can be recovered by BYHWT (P<0.01). The blood viscosity at high, medium, and low shear stress and the plasma viscosity in the model group were higher than those in the normal group (P<0.01, P<0.05). Compared with the model group, BYHWT restored the whole blood viscosity under high, medium and low shear stress and plasma viscosity (P<0.05,P<0.01). The model group had shorter prothrombin time (PT), shorter thrombin time (TT), and higher fibrinogen (FIB) than the normal group (P<0.05, P<0.01). BYHWT improved the TT and reduced the FIB in the rats with blood stasis syndrome (P<0.01). The platelet aggregation rate induced by arachidonic acid (AA) and adenosine diphosphate (ADP) in the model group was higher than that in normal group (P<0.01) and BYHWT decreased the platelet aggregation rate of the rats with blood stasis syndrome (P<0.01). The results of scanning electron microscopy showed that the model group exhibited excessive platelet activation, obvious pseudopodia, and increased aggregation of platelets compared with the normal group, while platelet activation and aggregation were rare in the BYHWT group. The serum levels of MMP-9 and ICAM-1 in the model group were higher than those in the normal group (P<0.01), which were decreased in the BYHWT group (P<0.05, P<0.01). ConclusionThe SD rats with the syndrome of acute blood stasis induced by ice water bath combined with injection of epinephrine demonstrate obvious changes in platelet function and morphology, inflammation, and abnormal cell adhesion. In the treatment of acute blood stasis in rats, BYHWT may reduce thrombosis and improve blood consistency and cohesion by mitigating inflammation, down-regulating cell adhesion factor overexpression, and improving platelet shape and function.
10.Establishment and Evaluation of Rat Model of Acute Coronary Syndrome with Combined Blood Stasis and Poison
Yue YOU ; Gaojie XIN ; Zixin LIU ; Lingmei LI ; Hongxu MENG ; Yue SHI ; Yanlei MA ; Ziyan WANG ; Jianxun LIU ; Lei LI ; Jianhua FU
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(19):49-60
ObjectiveTo compare the feasibility of establishing the rat model of acute coronary syndrome with combined blood stasis and poison by lipopolysacharide (LPS) injection, ligation of coronary artery and different combinations of the two methods. MethodA total of 225 male SD rats were randomly divided into sham operation group, simple coronary artery ligation group, first injected LPS group [LPS(5 mg·kg)injection 24 h before coronary artery ligation] and follow injected LPS group [LPS(5 mg·kg)injection 10 min after coronary artery ligation]. The indexes of each group were detected at 3, 24, 72 h after modeling, and the model was comprehensively evaluated. The general state and macroscopic evaluation indexes of traditional Chinese medicine (TCM) syndrome (tongue and pulse) of rats in each group were observed. ECG and echocardiography were used to evaluate cardiac function, and the myocardial ischemia and infarction areas were measured by Evans blue/2,3,5-triphenyltetrazolium chloride (TTC) staining. The content of creatine kinase isoenzyme (CK-MB), lactate dehydrogenase (LDH), creatine kinase (CK), and troponin T (cTnT) in serum as well as interleukin-1 β (IL-1β) and IL-6 changes were determined by biochemical method or enzyme-linked immunosorbent assay (ELISA). Hematology analyzer was adopted to determine the white blood cell (WBC) count, and the four coagulation indexes, platelet aggregation rate, hemorheology and other coagulation evaluation indexes were also detected. The myocardial tissue was observed by hematoxylin-eosin(HE)staining. ResultAfter 3 h of modeling, compared with the conditions in sham operation group, the R, G and B values of tongue of rats (P<0.01), pulse amplitude (P<0.01), and cardiac function in simple coronary artery ligation group were decreased, and the color of hypoglossal veins became purple(P<0.01). The content of CK, LDH, cTnT, IL-1β and IL-6 in serum(P<0.05), myocardial infarction area(P<0.01), and total number of WBCs (P<0.05)were increased. Compared with simple coronary artery ligation group, first injected LPS group and follow injected LPS group had increased hypoglossal veins, decreased R value of tongue and elevated cTnT content (P<0.01), while follow injected LPS group had reduced B value of tongue, decreased cardiac output (CO)(P<0.05), increased IL-1β content, and thinned left ventricular anterior walls at end-systole (LVAWs)(P<0.01). After 24 h of modeling, compared with sham operation group, simple coronary artery ligation group presented significantly decreased R, G and B values of tongue, lengthened purplish dark hypoglossal veins (P<0.01), reduced pulse amplitude(P<0.01) and cardiac function, enlarged myocardial infarction area(P<0.01), increased whole blood viscosity, platelet aggregation rate, fibrinogen (FIB), shortened prothrombin time (PT) and thrombin time (TT)(P<0.01), and elevated total number of WBCs (P<0.01)and content of CK, LDH, cTnT and IL-6 in serum(P<0.05). Compared with the conditions in simple coronary artery ligation group, the pulse amplitude, R, G and B values of tongue (P<0.01), and ejection fraction (EF) and fractional shortening (FS) scores (P<0.05)dropped, and hypoglossal veins were deepened and lengthened(P<0.05), and cTnT content was increased(P<0.01)in first injected LPS group and follow injected LPS group. However, follow injected LPS group had thinned LVPWs, increased LDH content, platelet aggregation rate(P<0.05), myocardial infarction area, and total number of WBC, level of IL-1β(P<0.05), and shortened TT(P<0.01). Additionally, 72 h after modeling, compared with sham operation group, simple coronary artery ligation group showed significantly reduced pulse amplitude, lowered R, G and B values of tongue, thickened and lengthened hypoglossal veins(P<0.01), decreased cardiac function, and increased content of cTnT, FIB, whole blood viscosity(P<0.01),platelet aggregation rate, level of IL-6 and IL-1β(P<0.05). Compared with the conditions in simple coronary artery ligation group, the hypoglossal veins of the first injected LPS group and the follow injected LPS group were more purple, and the content of cTnT was boosted(P<0.01), whereas follow injected LPS group had decreased pulse amplitude, R, G and B values of tongue, EF and FS scores (P<0.05), and enlarged myocardial infarction area(P<0.01). ConclusionCompared with the other modeling methods and models at different modeling time, the established model by LPS injection 10 min after coronary artery ligation for 24 h was more consistent with the clinical characteristics of acute coronary syndrome with combined blood stasis and poison.

Result Analysis
Print
Save
E-mail