1.Protective Effect of Wutou Chishizhi Wan on Vascular Endothelial Cells and Oxidative Stress in Myocardial Ischemia-reperfusion Rats
Meng-ni WANG ; Lu-lu XIE ; Zhao-peng ZHANG ; Bing LING ; Zhi-hui LIN ; Pai LIU ; Jun-peng GUO ; Hong-yan LIU
Chinese Journal of Experimental Traditional Medical Formulae 2020;26(21):40-47
Objective:To investigate the protective effect of Wutou Chishizhi Wan on myocardial ischemia reperfusion injury (MIRI) in rats, and observe its effect on such mechanisms as coagulation function, vascular endothelial cells and oxidative stress in rats. Method:A total of 40 SD rats were randomly divided into normal group, model group, positive drug group (Urokinase group) and Wutou Chishizhi Wan group, with 10 rats in each group. Except for the normal group, rat myocardial ischemia-reperfusion injury models were established. The changes of heart rate (HR) at 10 min before ischemia, 30 min after ischemia and 30, 60, 120 min (T0,T1,T2,T3,T4), and the change of electrocardiogram (ECG) J point after modeling in rats were observed. The pathological changes of rat myocardial tissue were observed by hematoxylin-eosin (HE) staining. The changes of four indexes of coagulation [prothrombin time (PT), activated partial thromboplastin time (APTT), thrombin time (TT), fibrinogen content decreased significantly (FIB)] in rats were observed. The contents of endothelin-1 (ET-1), thromboxane A2 (TXA2) and prostacyclin (PGI2) in serum and myocardium levels of superoxide dismutase (SOD), malondialdehyde (MDA) and glutathione peroxidase (GSH-Px) of MIRI rats were observed. Western blot assay was used for the detection of oxidative stress protein Keap1 and transcription factor-E2-related factor (Nrf2) expression levels in rat myocardial tissue. Result:Compared with the normal group, the ECG of MIRI rats showed significant myocardial ischemic injury-like changes, ST segment was significantly elevated, J point was significantly increased, and the incidences of HR in T1, T2, T3 and T4 were significantly reduced (
2.Effect of Yisui Jiedu Prescription on Apoptosis of Hippocampal Neurons Based onPI3K/Akt/Bad Signal Pathway in Rats with Vascular Dementia
Ling GAO ; Jian-jun ZHAO ; Ya-nan HU ; Li SU ; Ping LI ; Bing-mei WANG
Chinese Journal of Experimental Traditional Medical Formulae 2020;26(18):58-63
Objective:To investigate the effect of Yisui Jiedu prescription on hippocampal neuron damage in vascular dementia (VD) rats and to regulate phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt)/recombinant Bcl-2 associated death promoter (Bad) mechanisms of signaling pathways of neuronal apoptosis. Method:The 40 SD rats were divided into sham operation group, model group, donepezil hydrochloride group and Yisui Jiedu prescription group, with 10 rats in each group.VD animal model was prepared by bilateral carotid artery permanent ligation (2-VO) method.The sham operation group and the model group were intragastrically administered with normal saline, the donepezil hydrochloride group was intragastrically administered with donepezil hydrochloride 0.52 mg·kg-1. The Yisui Jiedu prescription group was administered with Yisui Jiedu prescription (11.11 g·kg-1), 1 time/d . After 30 days, Morris water maze was used to test the learning and memory ability of rats, hematoxylin-eosin (HE) staining was used to observe the histomorphological structure of hippocampal CA1 region. Ultrasound of neuron in rat hippocampal CA1 region was observed by transmission electron microscopy (TEM).Real-time fluorescent quantitative(Real-time PCR) was used to detect the Akt, Bad mRNA expression.Western blot was used to detect the Akt, p-Akt and Bad protein expression in hippocampus. Result:Compared with sham operation group, the learning and memory ability of model group decreased significantly(
3.Study on the Effects of Ginseng Root Extract on Autophagy ,Proliferation and Cytokine Secretion of Mice Splenic Lymphocytes and Its Mechanism
Fangyu LI ; Bin QI ; Shuai BIAN ; Yue ZHAO ; Shuyan LU ; Jiawen WANG ; Daqing ZHAO
China Pharmacy 2020;31(21):2597-2602
OBJECTIVE:To study the effects of gin seng root extract on autophagy ,proliferation and cytokine of splenic lymphocyte of mice ,and to study its mechanism. METHODS :The splenic lymphocyte of mice were divided into blank control group,positive control group (concanavalin A ,5 μg/mL),different concentration of ginseng root extract groups (32,125,500 μg/mL,by lyophilized powder ). After 48 h of culture with the corresponding medicine ,acridine orange staining method was used to detect autophagy of splenic lymphocyte ;CCK-8 assay was used to detect the cell proliferation ;ELISA assay was used to determine the levels of IL- 4,IL-6 and TNF-α in cell culture;Western blotting method was used to detect the expression of LC 3B and Beclin- 1,as well as the phosphorylation of AMPK ,AKT and mTOR. RESULTS :Compared with blank control group ,32, 125,500 μg/mL ginseng root extract could increase the number of acidic autophagosomes in splenic lymphocytes of mice(P<0.05 or P<0.01);125,500 μg/mL Ginseng root extract could significantly enhance the survival rate of splenic lymphocytes,the levels of IL- 4,IL-6 and TNF-α and the transformation of LC3BⅠ to LC 3BⅡ,significantly increased the protein expression of Beclin- 1, quinacrine cationic liposomes for treating non-small cell @163.com lung cancer[J]. J Drug Target ,2015,23(3):232-243. the phosphorylation of AMPK protein ,and significantly reduced the phosphorylation level of AKT and mTOR proteins ,with statistical significance (P<0.05 or P<0.01). CONCLUSIONS :Ginseng root extract can induce autophagy of mice splenic lymphocytes,activate the activity of splenic lymphocytes ,regulate the secretion of cytokine by activating AMPK and inhibiting the activation of AKT which can inhibit the activity of mTOR ,so as to exert immune enhancement effect.
4. Grade Standard of Glycyrrhizae Radix et Rhizoma Pieces
Ji-yu GONG ; Ling ZHAO ; Li-ru ZHAO ; Sha-sha WANG ; Guang-zhi CAI
Chinese Journal of Experimental Traditional Medical Formulae 2019;25(22):154-158
Objective: On the basis of traditional experience identification,and appearance characteristics and intrinsic index components of Glycyrrhizae Radix et Rhizoma pieces,a comprehensive evaluation was carried out to explore the method of grading Glycyrrhizae Radix et Rhizoma pieces and establish grading standards. Method: Based on the investigation of 44 batches of Glycyrrhizae Radix et Rhizoma pieces,including their properties,glycyrrhizin content and ammonium glycyrrhizinate content,Glycyrrhizae Radix et Rhizoma pieces were classified according to the market grading to establish the grading standards. Result: Based on the above data,the Glycyrrhizae Radix et Rhizoma pieces were classified into four grades. First-class:average diameter>1.66 mm,average weight>0.54 g,glycyrrhizin content>1.10%,glycyrrhizic acid content>2.12%. Second-class:the average diameter is between 1.40-1.66 mm,the average weight is between 0.42-0.54 g,the content of glycyrrhizin is between 0.74%-1.10%,and the content of glycyrrhizic acid is between 1.95%-2.12%. Third-class:the average diameter is between 1.07-1.40 mm,the average weight is between 0.29-0.42 g,the content of glycyrrhizin is between 0.65%-0.74%,and the content of glycyrrhizic acid is between 1.88%-1.95%. Substandard:the average diameter<1.07 mm,average weight<0.29 g,content of glycyrrhizin<0.65%,content of glycyrrhizic acid<1.88%. Conclusion: This experiment combines the traditional experience and modern analysis method,in order to develop a scientific,reasonable and accurate classification method.
5.Association Between Ferroptosis and Central Nervous System Diseases and A Review of Traditional Chinese Medicine Intervention
Hao LIU ; Hanying XU ; Zhong SHI ; Fan YAO ; Ziyue TIAN ; Tianye LAN ; Weimin ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(5):246-256
Central nervous system (CNS) is a sophisticated system subject to complex regulation, which dominates the high-level neural activities of the human body. Due to its complex physiological structure and refined regulatory mechanism covering a variety of diseases, CNS is the place where many chronic, refractory and rare diseases occur. Nerve cell is the basic unit of CNS, and its dysfunction and death is the root cause of CNS diseases. Ferroptosis is a new form of programmed cell death proposed in recent years, and has been proved to be closely related to the production and development of multiple CNS diseases. Traditional Chinese medicine (TCM), including Chinese herbs, acupuncture and moxibustion, and massage, has shown unique advantages in the treatment of CNS diseases for a long time. A large number of studies have demonstrated that TCM participates in the regulation of CNS diseases via regulating ferroptosis and shows a good research prospect. This paper summarized the characteristics of ferroptosis and discussed the association between ferroptosis and CNS diseases in pathological mechanism. We also reviewed the regulation of various CNS diseases by different TCM interventions through ferroptosis, providing references for TCM to participate in the treatment of CNS diseases properly in the future.
6.Optimization of the Honey-stir-fired Technology of Chelidonium majus
Renguang WANG ; Xintong SUI ; Yanqun WANG ; Xinrong WANG ; Xiujun ZHONG ; Tianyi ZHAO ; Shumin WANG
China Pharmacy 2021;32(12):1473-1479
OBJECTIVE:To optimize the honey-stir-fried technology of Chelidonium majus . METHODS :Taking the mass ratio of water to honey ,the ratio of honey water to C. majus ,stir-fired temperature ,stir-fired time as the factors ,the total contents of chelidonine ,coptisine hydrochloride ,sanguinarine,berberine,chelerythrine as response values ,Box-Behnken response surface method was used to optimize the processing technology ,and valifation test was conducted. RESULTS :The optimum process conditions were as follows the ratio of water to refined honey 1∶1.9(g/g),the ratio of honey water to C. majus 21∶100(g/g), stir-fried temperature 122 ℃,stir-fried time 10.40 min. After 3 times of validation ,average total contents of 5 components was 10.37 mg/g(RSD=0.23%),relative error of which with predicted value (10.39 mg/g)was 0.19%. CONCLUSIONS :The optimized honey-stir-fried technology of C. majus is stable and feasible.
7.Investigation of Adsorption and Desorption Properties of Total Ginsenosides on Anion-exchange Resins and Its LC-MS Analysis
Fa-jie WU ; Lin SUN ; Mei ZHAO ; Zhi-dong QIU ; Wei-nan WANG
Chinese Journal of Experimental Traditional Medical Formulae 2021;27(13):127-136
Objective:To compare the adsorption and desorption properties of different anion exchange resins for total ginsenosides, clarify their adsorption/desorption mechanism, and establish a simple protocol for the purification of total ginsenosides. Method:The adsorption and desorption properties of five different resins (D301, D315, D312, D330, D201) on total ginsenosides were evaluated with specific adsorption capacity, specific desorption capacity, desorption rate and recovery rate as indices. The adsorption kinetics and thermodynamics of the selected resin and D101 macroporous resin were investigated by pseudo-first-order and pseudo-second-order kinetic models, as well as Langmuir and Freundlich isothermal adsorption models, and the differences of adsorption mechanism between anion exchange resin and conventional macroporous resin were elucidated. The dynamic adsorption and desorption experiments were used to determine the optimum chromatographic parameters for anion exchange resin. After verifying the purification process of total ginsenosides, nine individual ginsenosides were qualitatively and quantitatively analyzed by liquid chromatography-mass spectrometry (LC-MS). Result:D301 anion exchange resin was obviously superior to the other four kinds of anion exchange resin, the optimum parameters were set as follows:pH 8 of loading solution, loading volume of 2 BV, loading speed of 4 BV·h-1, eluted with 3 BV of water and 20% ethanol for the impurities, eluted with 8 BV of 80% ethanol with elution speed of 4 BV·h-1. After purified by D301 resin, the enrichment coefficients of 9 monomer ginsenosides were simultaneously increased to different degrees, the overall enrichment coefficient was up to 5.3, the recovery rate for the total amount of these ginsenosides was calculated to be 80.9%, and the purity of total ginsenosides in Ginseng Radix et Rhizoma extract increased from 17.07% to 91.19%. Conclusion:D301 anion exchange resin is suitable for rapid and practical purification of total ginsenosides, hence allowing for the enrichment of high-purity total ginsenosides from Ginseng Radix et Rhizoma via one-dimensional column chromatography.
8.Analysis of analogical function acupoints for mental diseases based on.
Chinese Acupuncture & Moxibustion 2016;36(12):1331-1334
OBJECTIVETo summarize the acupoints for mental diseases in(GB/T30233-2013) so as to provide theoretical evidence for acupoint compatibility and acupuncture-moxibustion prescription.
METHODSThe acupoints for mental diseases inwere artificially retrieved,and then their frequencies,meridians and locations were analyzed.
RESULTSEighty acupoints were collected totally. Shenmen (HT 7) was with the highest frequency,and Baihui (GV 20) was followed. The frequently-used meridians in turn were governor vessel (GV),bladder meridian of foot-,small intestine meridian of hand-and stomach meridian of foot-,etc.,among which GV showed the highest number with 17 (21.25%) acupoints. As for the positions of analogical function acupoints,they in turn were head and face,upper limbs,lower lumbs,back and waist,chest and abdomen,with 30 percent acupoints locating at head and face.
CONCLUSIONSAnalogical acupoints for mental diseases are focused at head and face,and they are mainly along GV. The treatment is based on syndrome differentiation.
9.Effect of Modified Shenhong Tongluo Prescription on Cell Apoptosis in Rats with Myocardial Ischemia-reperfusion Injury by Bcl-2/Bax/Caspase-3 Signaling Pathway
Yingrong LAI ; Qianlin ZHAO ; Lihong JIANG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(11):104-110
ObjectiveTo investigate the mechanism of modified Shenhong Tongluo prescription on cell apoptosis in rats with myocardial ischemia-reperfusion injury (MIRI). MethodSixty Sprague-Dawley (SD) rats were randomly divided into a blank group, a model group, low-, medium-, and high-dose groups of modified Shenhong Tongluo prescription, and a simvastatin group. Except for the blank group, a rat model of MIRI was prepared by ligating the left anterior descending coronary artery. Starting from the first day after successful modeling, the blank group (1.0 mL·kg-1 physiological saline), model group (1.0 mL·kg-1 physiological saline), low-, medium-, and high-dose groups of modified Shenhong Tongluo prescription (1.031, 2.063, and 4.126 g·kg-1 Shenhong Tongluo prescriptiona standard concentrate), and simvastatin group (0.71 mg·kg-1 simvastatin) were orally administered once daily for 2 weeks. Hematoxylin-eosin (HE) staining was used to observe the pathological changes of cardiomyocytes. Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of serum creatine kinase isoenzyme (CK-MB), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α). TdT-mediated dUTP nick-end labeling(TUNEL) staining was used to detect the apoptosis rate of rat cardiomyocytes. Western blot was used to detect the expression levels of apoptosis-related proteins B-cell lymphoma-2 (Bcl-2), Bcl-2-associated X protein (Bax), and caspase-3. ResultCompared with the blank group, in the model group, HE staining showed disturbed arrangement of cardiomyocytes, incomplete fibers, focal necrosis of cardiomyocytes, and inflammatory cell infiltration; serum CK-MB, IL-6, and TNF-α levels were significantly increased (P<0.05); apoptosis rate of cardiomyocytes was significantly increased (P<0.01), with significantly increased expression levels of Bax and Caspase-3 proteins, and significantly decreased Bcl-2 expression (P<0.05). Compared with the model group, the low-, medium-, and high-dose groups of modified Shenhong Tongluo prescription significantly reduced CK-MB, IL-6, and TNF-α levels (P<0.05), significantly downregulated cardiomyocyte apoptosis rate (P<0.05), significantly decreased Bax and Caspase-3 proteins, and significantly increased Bcl-2 expression levels (P<0.01). In the modified Shenhong Tongluo prescription groups, the expression levels of Bax and Caspase-3 proteins significantly decreased with increasing dosage, while the expression level of Bcl-2 significantly increased with increasing dosage of modified Shenhong Tongluo prescription (P<0.05). ConclusionShenhong Tongluo prescription can alleviate myocardial tissue pathological damage and reduce myocardial cell apoptosis, possibly by inhibiting Caspase-3 and Bax expression and promoting Bcl-2 expression.
10.Prevention and therapeutic effects of sika deer velvet collagen hydrolysate on osteoporosis in rats by retinoic acid.
Yinqing LI ; Yu ZHAO ; Xiaodi SUN ; Xiaobo QU
China Journal of Chinese Materia Medica 2010;35(6):759-762
The objective was to evaluate the preventive and therapeutic effects of the collagen hydrolysate extracted from Sika deer velvet (CSDV) on osteoporosis rats induced by retinoicacid. Histomorphometric indices and serum biochemical parameters were measured in osteoporosis rats treated with/without antler collagen and in sham-operated rats. Our results were as follows: compared with the osteoporosis group, significant elevation in the levels of bone mineral density (BMD), Ca, P and static histomorphometric indexes and biomechanical properties, but reduction in the level of serum alkaline phosphatase (ALP) were observed in antler collagen-treated groups. However, the above function with the collagenase solution velvet material varied with the different doses. In conclusion, the extracted collagen is found to play a role in the prevention and treatment of osteoporosis rats by retinoic acid.
Animals
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Bone Density
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drug effects
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Collagen
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metabolism
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therapeutic use
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Female
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Humans
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Male
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Osteoporosis
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prevention & control
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Osteoporosis, Postmenopausal
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Rats
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Rats, Wistar
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Tretinoin
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therapeutic use