Study on regulation of CYP450 enzyme system to reduce liver toxicity through compatibility of Aconiti Kusnezoffii Radix Cocta with Chebulae Fructus and Glycyrrhizae Radix et Rhizoma / 中国中药杂志

Aconiti Kusnezoffii Radix Cocta is one of the most commonly used medicinal materials in Mongolian medicine. Due to the strong toxicity of Aconiti Kusnezoffii Radix Cocta, Mongolian medicine often uses Chebulae Fructus, Glycyrrhizae Radix et Rhizoma to reduce the toxicity, so as to ensure the curative effect of Aconiti Kusnezoffii Radix Cocta while ensuring its clinical curative effect, but the mechanism is not clear. The aim of this study was to investigate the effects of Chebulae Fructus, Glycyrrhizae Radix et Rhizoma and Aconiti Kusnezoffii Radix Cocta on the mRNA transcription and protein translation of cytochrome P450(CYP450) in the liver of normal rats. Male SD rats were randomly divided into negative control(NC) group, phenobarbital(PB) group(0.08 g·kg~(-1)·d~(-1)), Chebulae Fructus group(0.254 2 g·kg~(-1)·d~(-1)), Glycyrrhizae Radix et Rhizoma group(0.254 2 g·kg~(-1)·d~(-1)), Aconiti Kusnezoffii Radix Cocta group(0.254 2 g·kg~(-1)·d~(-1))and compatibility group(0.254 2 g·kg~(-1)·d~(-1),taking Aconiti Kusnezoffii Radix Cocta as the standard). After continuous administration for 8 days, the activities of total bile acid(TBA), alkaline phosphatase(ALP), amino-transferase(ALT) and aspartate aminotransferase(AST)in serum were detected, the pathological changes of liver tissue were observed, and the mRNA and protein expression levels of CYP1 A2, CYP2 C11 and CYP3 A1 were observed. Compared with the NC group, the serum ALP, ALT and AST activities in the Aconiti Kusnezoffii Radix Cocta group were significantly increased, and the ALP, ALT and AST activities were decreased after compatibility. At the same time, compatibility could reduce the liver injury caused by Aconiti Kusnezoffii Radix Cocta. The results showed that Aconiti Kusnezoffii Radix Cocta could inhibit the expression of CYP1 A2, CYP2 C11 and CYP3 A1, and could up-regulate the expression of CYP1 A2, CYP2 C11 and CYP3 A1 when combined with Chebulae Fructus and Glycyrrhizae Radix et Rhizoma. The level of translation was consistent with that of transcription. The compatibility of Chebulae Fructus and Glycyrrhizae Radix et Rhizoma with Aconiti Kusnezoffii Radix Cocta could up-regulate the expression of CYP450 enzyme, reduce the accumulation time of aconitine in vivo, and play a role in reducing toxicity, and this effect may start from gene transcription.

Inhibitory Effect and Mechanism of Jingulian Extract on LPS-induced RAW264.7 Cell Inflammatory Response Based on PI3K/Akt Signaling Pathway / 中国实验方剂学杂志

Objective:To explore the inhibitory effect and mechanism of Jingulian extract （JGL） on inflammation. Method:The following groups were set up in this study： a control group （10% fetal bovine serum）， a lipopolysaccharide （LPS） model group （0.5 mg·L^-1）， and JGL groups （10， 20， 40， 60， 80， 120， 160， 200， 250， 300 mg·L^-1 + 0.5 mg·L^-1LPS）. The RAW264.7 cells were cultured for 24 hours. Cell proliferation was detected by cell counting kit-8 （CCK-8） assay. Nitric oxide （NO） release was detected by Griess assay. The release of cytokines interleukin （IL）-1<italic>β</italic>， IL-6， IL-10， and tumor necrosis factor （TNF）-<italic>α</italic> was determined by enzyme linked immunosorbent assay （ELISA）. The expression of inducible nitric oxide synthase （iNOS） and intraprostaglandin peroxidase synthase 2 （PTGS2）/cyclooxygenase-2 （COX-2） was measured by real-time fluorescence-based quantitative polymerase chain reaction （Real-time PCR） and the activation of key proteins in the phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt） signaling pathway by Western blot. Result:Compared with the control group， LPS （0.5 mg·L^-1）could promote the proliferation of RAW264.7 cells after stimulation for 24 hours （<italic>P</italic><0.01）. Compared with the model group， JGL had no significant effect on cell proliferation. Compared with the control group， LPS （0.5 mg·L^-1）increased the release of NO， IL-1<italic>β</italic>， IL-6， IL-10， and TNF-<italic>α</italic> （<italic>P</italic><0.01）. Compared with the model group， JGL （20-300 mg·L^-1）inhibited the release of NO in a dose-dependent manner after stimulation for 24 hours （<italic>P</italic><0.05） and reduced IL-1<italic>β</italic>， IL-6， and IL-10 （<italic>P</italic><0.05， <italic>P</italic><0.01）， but no obvious inhibition on the release of TNF-<italic>α</italic> was observed. LPS （0.5 mg·L^-1） could induce the expression of iNOS and PTGS2/COX-2 genes as compared with the control group （<italic>P</italic><0.05， <italic>P</italic><0.01）. JGL could down-regulate the mRNA expression of iNOS and PTGS2/COX-2 genes as compared with the model group （<italic>P</italic><0.05， <italic>P</italic><0.01）. LPS （0.5 mg·L^-1） could activate the PI3K/Akt pathway （<italic>P</italic><0.01） as compared with the control group， while JGL （10， 20， 40， and 80 mg·L^-1） decreased the expression of PI3K-p110， p-p85， and p-Akt （<italic>P</italic><0.01）， and inhibited the activation of PI3K/Akt pathway. Conclusion:JGL extract could significantly inhibit the inflammatory response and activation of the PI3K/Akt pathway induced by LPS in RAW264.7 cells. The anti-inflammatory effect was related to the inhibition of the PI3K/Akt pathway.

Attenuation Mechanism of Aconti Lateralis Radix Praeparata Combined with Glycyrrhizae Radix et Rhizoma Based on Heart CYP450 Isozymes / 中国实验方剂学杂志

<b>Objective::To investigate the effect of Aconiti Lateralis Praeparata Radix combined with Glycyrrhizae Radix et Rhizoma on mRNA expression of cytochrome P450 (CYP450) enzymes in rat' s heart. <b>Method::Male Sprague Dawley rats were randomly divided into the control group, the inducer group (0.08 g·kg^-1·d^-1), the aconite group (0.5 g·kg^-1·d^-1), the licorice group (0.5 g·kg^-1·d^-1) and the compatibility group (aconite and licorice group 0.5 g·kg^-1·d^-1), with 6 rats in each group. In each group, the serum concentrations of aspartate transaminase (AST), creatine kinase (CK) andlactate dehydrogenase (LDH) were respectively measured. Hematoxylin-eosin(HE) staining was used to detect the pathologic changes in the heart tissue of the rats. The expression of several CYP genes were measured. <b>Result::After combination, the contents of AST, CK and LDH were lower than those of the aconite group. At the same time, the pathological results showed cardiac injury in rats in the aconite group, and licorice could alleviate the cardiac injury caused by aconite. At the level of gene transcription, the glycyrrhizae group could up-regulate the expressions of CYP2C11 and CYP2J3 to different degrees (<italic>P</italic><0.05), and the combination of licorice and aconite could up-regulate the expression of CYP2B1, CYP2C11 and CYP2J3 family (<italic>P</italic><0.05). It suggested that the combination could promote the formation of EETs in arachidonic acid(AA) metabolism, and the aconite could up-regulate the expressions of CYP4A1, CYP4A3, CYP4F5 and CYP4F6 in CYP4 family (<italic>P</italic><0.05), promote the formation of 20-hydroxyeicosatetraenoic acid(20-HETE). The combination of aconite and licorice can weaken the ability of aconite, down-regulate the expression of CYP4A3, CYP4F1, CYP4F5 and CYP4F6 family mRNA (<italic>P</italic><0.05), inhibit the formation of 20-HETE and reduce the cardiotoxicity caused by aconite. <b>Conclusion::The combination of licorice and aconite can regulate the expression of CYP450 enzyme in the heart, increase the production of EETs and the content of 20-HETE, and finally play a role in reducing toxicity.

Effect of Tetrandrine on Col-I and FN in TGF-β1-induced MRC-5 Cells / 中国实验方剂学杂志

Objective:To investigate the effect of tetrandrine on transforming growth factor-β1（TGF-β1）stimulated MRC-5 cells. Method:Different concentrations of TGF-β1 (0, 2.5, 5, 10, 20, 40 μg·L-1) were applied to MRC-5 cells. Proliferation toxicity of TGF-β1 to MRC-5 was detected by cell counting kit-8 (CCK-8) method. Detection of alpha smooth muscle actin (α-SMA) and Vimentin's expression levels in MRC-5 by Western blot. Detection of changes of collagen I(Col-I) and fibronectin (FN)'s expression levels in MRC-5 supernatants by enzyme linked immunosorbent assay(ELISA) kit. And the appropriate concentration of TGF-β1 activated MRC-5 cells was screened. The appropriate concentration of TGF-β1 and different concentrations of Tet (0, 2.5, 5, 10, 20, 40 μmol·L-1) were applied to MRC-5 cells, and CCK-8 method was used to screen safe concentration again. Western blot was used to detect changes in α-SMA and Vimentin expression levels in MRC-5 cells, and ELISA method to detect changes in Col-I and FN in MRC-5 cell supernatant. Result:Compared with the blank group, 20，40 μg·L-1 of TGF-β1 had toxic effects on MRC-5 cells at 24 hours (P<0.05), and 10，20，40 μg·L-1 of TGF-β1 had toxic effects on MRC-5 cells at 48 h (P<0.05).When Tet is added for 24 h, the half inhibitory concentration (IC50) value was 14.07 μmol·L-1, and when cultured for 48 h, the IC50 value was 7.51 μmol·L-1. Compared with the blank group, the relative contents of α-SMA, FN and Col-I in the 5 μg·L-1 of TGF-β1 group were obviously increased (P<0.05), and the relative contents of Vimentin were significantly increased (P<0.01), and the relative contents of FN and Col-I, α-SMA and Vimentin in 10 μg·L-1 group were significantly increased (P<0.01). 10 μg·L-1 of TGF-β1 was co-cultured with Tet at different concentrations. Compared with the TGF-β1 group, the relative levels of α-SMA, Vimentin and FN in the 5 μmol·L-1 of Tet group were significantly reduced (P<0.01), and the relative levels of Col-I were obviously reduced (P<0.05). In the Tet 10 μmol·L-1 group, the relative contents of the α-SMA, Vimentin, FN and Col-I were significantly reduced (P<0.01). Conclusion:TGF-β1 can increase the levels of Col-I, FN and other extracellular matrices in MRC-5 cells, and Tet can effectively inhibit the occurrence of this change. It is suggested that Tet may inhibit secreting extracellular matrix of fibroblasts in the formation of pulmonary fibrosis.

Toxicological Technology in Evaluation and Application of Traditional Chinese Medicine Healthcare Food / 中国实验方剂学杂志

China is rich in herbs that can be used as medicine and food. The application of this kind of traditional Chinese medicine (TCM) as raw material is the basis for guiding the development of TCM healthcare food. TCM healthcare food has advantages in keeping in good health and reducing the risk of disease. At the same time, with the implementation of " Healthy China 2030" , it is urgent to develop a series of herbal compound healthcare food with the characteristics of TCM to meet the national and social needs. With the development of scientific research, however, there is a lack of research on the scientific evaluation of the safety of TCM healthcare food raw materials, which results in insufficient risk assessment theory and technology of healthcare food at present. Therefore, the prediction and evaluation of the toxicity of important components in this kind of raw materials has become an urgent task of food safety. Toxicology plays an important role in human health risk assessment, and its sub-disciplines have been constantly emerging. Computational toxicology, monolayer culture cell model technology, in vitro induced activity screening engineering cell technology, microfluidic chip technology and in vivo toxicology technology have become important research tools for toxicity prediction and evaluation in this field. With the rapid development of toxicology technology, food safety and human life and health can be effectively guaranteed. It will effectively promote the safety production of healthcare food industry in China and guarantee the quality and safety of terminal products. It has a great social significance, plays a positive leading role in the technological progress of the whole industry, and generates considerable economic benefits. This paper reviews recent advances and applications of new toxicological methods and models for predicting and evaluating important ingredients in TCM healthcare food raw materials.

Effect of Effective Components of Salvia Miltiorrhiza on TGF-β1 Induced Interstitial Transformation of Pulmonary Epithelial Cells / 中国实验方剂学杂志

Objective::To screen out the effective components of Salvia miltiorrhiza by establishing an in vitro model of pulmonary epithelial mesenchymal transformation. Method::Different concentrations of salvianolic acid A (10, 20, 40, 80, 160 μmol·L-1), salvianolic acid B (10, 20, 40, 80, 160 μmol·L-1), tanshinol (10, 20, 40, 80, 160 μmol·L-1), tanshinoneⅡA (10, 20, 40, 80, 160 μmol·L-1) and the blank group were applied to A549 cell, cell proliferation and cytotoxicity assay (MTS) were used to detect the proliferation effect of menthol on A549 cells.After screening the safe concentration of the active ingredients of salvia miltiorrhiza by MTS, cells were divided into blank group, model group, salvianolic acid A group, salvianolic acid B group, tanshinol group and tanshinoneⅡA.Then, the inhibitory effect of the active ingredients of salvia miltiorrhiza on the proliferation of A549 cells induced by TGF-β1 was detected by MTS. Enzyme linked immunosorbent assay (ELISA) method to detect salvia miltiorrhiza effective component of fiber protein(FN), collagen type I (COL-Ⅰ) expression. Based on the above results, the active components of salvia miltiorrhiza, which have best inhibition were screened out, and their effects on the expression of E-calcium-viscosity (E-Cad) protein were detected by Western blot. Result::Compared with blank group, salvianolic acid A 40 μmol·L-1, salvianolic acid B 160 μmol·L-1, tanshinol 160 μmol·L-1 had toxic effects on A549 cells (P<0.05). In the non-toxic concentration range, compared with the model group, salvianolic acid A 10, 20 μmol·L-1, salvianolic acid B 80 μmol·L-1 showed inhibition effect after 24 h culture (P<0.05). After 72 h culture, salvianolic acid A 5, 10, 20 μmol·L-1, salvianolic acid B 40, 80 μmol·L-1inhibition effect was very significant (P<0.01). ELISA results showed that with the blank group, model group cells the expression of FN and COL-Ⅰ increased significantly (P < 0.01). Compare with model group, salvianolic acid A 20 μmol·L-1, salvianolic acid B 80 μmol·L-1 inhibited FN and COL-Ⅰ(P<0.05). Western blot results showed that salicylic acid A and salicylic acid B had protective effects on E-Cad (P<0.01). Conclusion::Salvianolic acid A and salvianolic acid B have inhibitory effects on epithelial mesenchymal transformation by TGF-β1, which may be the main effective components of salvianolic acid in the treatment of pulmonary fibrosis.

Research Progress in Pharmacology of Lianhua Qingwen Preparation / 中国实验方剂学杂志

Lianhua Qingwen preparation （LHQW） is a Chinese traditional patent medicine approved by China Food and Drug Administration （CFDA）， and divided into two dosage forms， namely capsules and granules. Based on TCM theory， its therapeutic functions are contagion-clearing， detoxification， antipyretic， and lung-ventilating regulation， with influenza as its indication. In this paper， its pharmacological activities were reviewed. LHQW had a significant anti-virus effect characterized by a broad-spectrum pattern. It was reported that it not only possessed definitely suppressive effect on a series of influenza viruses， respiratory syncytial virus， coxsackie， enterovirus， herpes simplex virus，but also displayed a significant inhibitory effect on both the new corona pneumonia virus （SARS-CoV-2） and SARS coronavirus （SARS-CoV）. Studies showed that LHQW has obvious anti-inflammatory effects on a variety of inflammation models. It can significantly increase the delayed hypersensitivity of immunocompromised mice （caused by hydrocortisone） against 2， 4-dinitrofluorobenzene， and improve their cellular immune function. It can improve the phagocytosis function of peritoneal macrophages， the serum hemolysin antibody level and the humoral immune function of mice with a low immune function， with a immunomodulatory effect. In addition， LHQW has therapeutic effects on the symptoms induced by respiratory tract infections， such as fever， cough and phlegm， so as to block the vicious circle of multiple pathological links of the disease， and bring the advantages of multi-target， multi-link and multi-approach overall treatment of TCM into play.

Commonly Used Vomiting Animal Models / 中国实验方剂学杂志

Vomiting is a common clinical symptom. Long-term severe vomiting could seriously reduce the life quality of patients, so it is necessary to be intervened with antiemetic drugs. Traditional Chinese medicine has a long history in the treatment of vomiting with remarkable a curative effect. It has the advantages of multi-component, multi-target and multi-system synergistic antagonism. However, due to the active substance and unclear mechanism, it is urgent to adopt an internationally recognized vomiting model system to evaluate the antiemetic characteristics, elucidate the vomiting mechanism, and provide reference for better clinical application. Therefore, this paper systematically introduces several vomiting animal models that are widely used at home and abroad. According to the authors' own experimental experience, this paper focuses on the rat and mice pica models for reference of relevant researchers. Specifically, ferrets are an internationally recognized ideal vomiting animal model, and the golden standard for evaluating the effects of antiemetic drugs, suncus murinus is the smallest mammalian vomiting model. Rodents have no vomiting reflexes, but studies have shown that its pica behavior is equivalent to the vomiting behavior of other species. Because the easy availability and operation, the model has been promoted and applied in mainland China. Guizhou mini-pig model is a self-developed medium-sized mammalian vomiting model with a similar anatomical structure and vomiting characteristics to human, and worthy of popularization and application. In conclusion, different vomiting models have their own characteristics that need to be optimized according to the purpose of experiments and samples.

Effect of Pudilan Xiaoyan Oral Liquid on Acute Lung Injury Rat Induced by Lipopolysaccharide / 中国实验方剂学杂志

Objective:To explore the effect and mechanism of Pudilan Xiaoyan oral liquid(PDL) on the acute lung injury rat induced by lipopolysaccharide (LPS). Method:The 72 Wistar rats were randomly divided into control group, model group, dexamethasone group, PDL 7, 3.5, 1.75 g·kg-1·d-1 group according to body weight.The acute lung injury model was made through inhalation with lipopolysaccharide in the model group, hexadecadrol group, PDL 7, 3.5, 1.75 g·kg-1·d-1 group.To examining each rat alveolar lavage fluid (BALF) of the total number of white blood cells, enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of nuclear transcription factors-kappa B (NF-κB) and interleukin-10 (IL-10).Hematoxylin-eosin(HE) staining was used to observe morphological changes of lung tissue and explore different doses of PDL effect on acute lung injury in rats. Result:Compared with model group, the account of leukocyte in BALF decreased significantly in PDL 7 g·kg-1·d-1 group and PDL 3.5 g·kg-1·d-1 group (PκB significantly decreased in PDL 7, 3.5, 1.75 g·kg-1·d-1 group (P-1·d-1 group (P-1·d-1 group, the inflammation, edema and congestion in lung tissue reduced (PConclusion:PDL has a significant protective effect on the inflammation of acute lung injury model, and its mechanism is related to the expressions of NF-κB and IL-10. PDL could also repair the injury of lung in acute lung injury model.

Pharmacodynamic Evaluation of Tanreqing Inhalation Solution / 中国实验方剂学杂志

Objective: To clarify the antitussive, expectorant, antipyretic and anti-inflammatory effects of Tanreqing inhalation solution, and provide basis and data support for further research and development of this preparation. Method: The methods of cough induced by ammonia and tracheal phenol red excretion were used to observe the antitussive and expectorant effects of Tanreqing inhalation solution in mice. The fever model of rats was established by intraperitoneal injection of bacterial lipopolysaccharide(LPS) to observe the antipyretic effect of the Tanreqing inhalation solution, the acute pneumonia model of rats was established by atomizing LPS inhalation, and the anti-inflammatory effect of Tanreqing inhalation solution was observed. Result: Tanreqing inhalation solution could reduce the number of coughs in mice induced by ammonia water, increase the amount of phenol red excretion in mouse trachea, decrease the levels of body temperature and its related regulatory factors of prostaglandin E2(PGE2) and cyclic adenosine monophosphate(cAMP) of rats induced by LPS, decrease the white blood cell(WBC) count and the neutrophil ratio(NEUT) in bronchoalveolar lavage fluid(BALF) of rats with LPS-induced acute pneumonia, and reduce the levels of nuclear transcription factor-κB(NF-κB) and interleukin-1β(IL-1β) in lung tissue. Conclusion: Tanreqing inhalation solution has obvious antitussive, expectorant, antipyretic and anti-inflammatory effects, which is worthy of further development and promotion.

Effect of Different Administration Methods with Bleomycin on Pulmonary Fibrosis in Rats / 中国实验方剂学杂志

Objective:Compare the effects of 3 administration methods (tracheal perfusion, tail vein injection and aerosol inhalation) with bleomycin (BLM) in inducing pulmonary fibrosis in rats, in order to find out the optimal administration methods. Method:Eighty sprague-dawley (SD) male rats with SPF were randomly divided into aerosol inhalation blank group, single tracheal perfusion group(10 mg·kg-1), multiple tracheal perfusion group(5 mg·kg-1), single intravenous injection group(150 mg·kg-1), multiple intravenous injection group(50 mg·kg-1), single aerosol inhalation group (30 min)and multiple aerosol inhalation group(30 min). The mortality and body weight of rats in each group were observed at 7 d, 14 d and 28 d after the administration. And 28 days later after the administration, the lung coefficients of rats in each group were observed, paraffin sections were prepared, hematoxylin-eosin staining (HE) and Masson staining were performed, and the contents of hydroxyproline (HYP) and plasminogen activator inhibitor-1 (PAI-1) in lung tissues were detected by enzyme-linked immunosorbent assay (ELISA), so as to evaluate the alveoli inflammation and pulmonary fibrosis of rats in each group. Result:Compared with the aerosol inhalation blank group, the rats in the trachea perfusion group had the highest mortality among the drug treatment groups. The pulmonary coefficients of rats in the multiple intravenous injection group and the multiple inhalation group were significantly higher than those in the blank group(PPPConclusion:Bleomycin was inhaled repeatedly to establish pulmonary fibrosis model. The pathological injury and physiological indexes of the model rats were relatively stable, which conforms with the evolution process of pulmonary fibrosis.

Anti-diarrhea Effect and Mechanism of Traditional Chinese Medicine / 中国实验方剂学杂志

Diarrhea is a common clinical digestive tract disease, and the second major cause of illness and death among children. At present, long-term irrational use of chemical drugs and antibiotics in the treatment of diarrhea will accelerate the emergence of drug resistance of intestinal pathogenic bacteria, and modern drug preparations and therapies cost very high, which therefore bring difficulties to clinical treatment. As a pure natural drug containing a variety of active ingredients, traditional Chinese medicine(TCM) has the advantages of low residue, low toxicity and low drug resistance. And the study shows that TCM preparations, single TCM and effective components of TCM have a unique efficacy in the treatment of diarrhea, which makes anti-diarrhea TCM become a hotspot in the research field of anti-diarrhea drugs. Therefore, through consultation of relevant Chinese and foreign literatures in recent years, this study summarizes relevant anti-diarrhea TCM, and systematically expounds the treatment of diarrhea with TCM in the aspects of regulating the balance of water and electrolyte, anti-inflammation, interfering with gastrointestinal hormone secretion, protecting intestinal mucosa, promoting intestinal repair. Specifically, because the excessive secretion and absorption of liquids and electrolytes by intestinal epithelial cells are two important factors in the pathogenesis of diarrhea, the reduction of intestinal fluid secretion and the promotion of intestinal fluid absorption become effective means for the treatment of diarrhea. This paper reviews the effects and mechanisms of TCM against diarrhea, in order to provide theoretical support for the study of the anti-diarrhea mechanism and the research and development of new drugs of TCM against diarrhea.

Mechanism of Dichroa Alkali Salt Inducing Vomiting Based on Pica in Mice / 中国实验方剂学杂志

Objective: To study the possible mechanism of dichroa alkali salt (DAS) in inducing vomiting. Method: Mice pica model was used to observe the antagonistic effect of the three different kinds of antiemetic drugs[dopamine receptor antagonist metoclopramide, 5-hydroxytryptamine 3 (5-HT3) receptor antagonist ondansetron and neurokinin-1 receptor antagonist aprepitant] on body mass, food intake, kaolin consumption, diarrhea and death induced by DAS to preliminarily clarify the possible pathogenic pathway of DAS. Then, the expression of 5-HT and substance P(SP) in ileum and medulla of mice induced by DAS alone at different time points was detected by enzyme-linked immunosorbent assay to confirm whether DAS could affect the changes of these two neurotransmitters. Result: After treatment with ondansetron and aprepitant, DAS-induced reduction in food intake of mice was significantly improved on the 4th day after continuous administration and on the 1st day after drug administration (Prd day after administration, DAS-induced body mass loss of mice was significantly improved (PConclusion: The mice pica model can be used to effectively characterize DAS-induced vomiting. DAS-induced pica in mice may be associated with the increase of 5-HT and SP in ileum and medulla. Ondansetron and aprepitant can effectively antagonize DAS-induced pica in mice.

Investigation of Physicochemical Properties and Determination Methods of Liposomal Entrapment Efficiency of Dichroa Alkali Hydrochloride / 中国实验方剂学杂志

Objective: To study on the physical and chemical properties of dichroa alkali hydrochloride and to establish a method for the determination of entrapment efficiency of dichroa alkali hydrochloride liposomes. Method: HPLC was used to determine the content of dichroa alkali hydrochloride with mobile phase of acetonitrile-water-triethylamine-glacial acetic acid(9:91:0.35:0.75) and detection wavelength at 265 nm.The oil-water partition coefficient of this compound in different pH range was measured by shake flask method.The stability of the dichroa alkali hydrochloride in phosphate buffer solution with different pH after sterilization at 125℃ for 30 min was investigated.Ammonium sulfate gradient method was used to prepare dichroa alkali hydrochloride liposomes.The microcolumn was prepared by dextran gel and cation exchange resin,respectively.Then the free drug and liposome were separated by centrifugation,the drug content was measured,and the encapsulation efficiency was calculated.The t-test was performed using SPSS 20.0 software,the differences between these two methods were compared. Result: In the pH 6-9,the oil-water partition coefficient of dichroa alkali hydrochloride increased with increasing of pH,which was between 0.016 and 1.44;the recovery rate of dichroa alkali hydrochloride after sterilization was 37.16%-57.91%.Between the dextran gel microcolumn centrifugation and the cation exchange resin microcolumn centrifugation,there was no significant difference in the entrapment efficiency of the liposomes. Conclusion: Dichroa alkali hydrochloride is suitable for preparation of liposomes.However,its stability is not ideal,so the experimental temperature should be strictly controlled in the preparation process.Dextran gel microcolumn centrifugation and cation exchange resin microcolumn centrifugation can be used to determine the entrapment efficiency of dichroa alkali hydrochloride liposomes,and the cation exchange resin microcolumn centrifugation is suggested after comparison.

Protective Effect of Renshen Sinitang and Its Active Ingredients on Myocardial Cell Injury Induced by Pentobarbital Sodium / 中国实验方剂学杂志

Objective: To explore the protective effect and mechanisms of Renshen Sinitang and its active ingredients on cardiomyocyte injury induced by pentobarbital sodium. Method: H9C2 cells were sub-cultured with ginsenoside Rb2 0.01, 0.1, 1 μmol ·L-1, Re 0.01, 0.1, 1 μmol·L-1, isoliquiritigenin 20, 40, 80 μmol·L-1, glycyrrhetinic acid 10, 20, 40 μmol·L-1, Renshen Sinitang, 10, 100, 400 mg·L-1, for 4 h. After treatment with 0.1% of sodium pentobarbital for 30 min, cell viability, lactate dehydrogenase (LDH), lipid peroxide malondialdehyde (MDA), Na+-K+-adenosine triphosphate(ATP) ase, Ca2+-ATPase activity, and real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) were used to detect the expressions of peroxisome proliferative activated receptor-1α (PGC-1α), B-cell lymphoma-2 associated X protein(Bax) and cysteine aspartate-specific protease-3(Caspase-3) mRNA. Result: Renshen Sinitang and its active ingredients have a protective effect on heart failure cell model. Compared with the normal group, the cell survival rate of the model group decreased significantly, while the LDH and MDA contents increased significantly, and the Na+-K+-ATPase activity increased. Ca2+-ATPase activity was significantly decreased, PGC-1α mRNA expression was down-regulated, Bax and Caspase-3 mRNA expressions indicates the modeling(P+-K+-ATPase activity, increased Ca2+-ATPase activity, up-regulated PGC-1α mRNA expression, and inhibited Bax and Caspase-3 mRNA expression (PPConclusion: Renshen Sinitang and its active ingredients have a significant protective effect on heart failure cell model, and its mechanisms of action are related to anti-oxidation, improvement of mitochondrial energy metabolism and inhibition of mitochondrial apoptosis pathway.

Study on main pharmacodynamics and underlying mechanisms of 999 Ganmaoling / 中国中药杂志

To observe synergistic effects of 999 Ganmaoling (GML) and its Chinese/Western materia medica (CMM and WMM) on pharmacodynamic action and to study underlying mechanisms, their anti-inflammatory, antipyretic effects were compared by assaying the increased capillary permeability induced by glacial acetic acid in mice, ear swelling induced by Xylene in mice, non-specific pleurisy induced by carrageenan in rats, and yeast induced fever in rats. Crystal violet (CV) and microbial activity (XTT) assay were used to evaluate the inhibition of GML and its CMM and WMM on KPN biofilm formation, and scanning electron microscopy (SEM) was applied for observing KPN biofilm morphology changes. The results showed that compared with control group, GML could reduce exudation amount of Evans-Blue and the degree of Ear swelling significantly, and CMM and WMM have no significant effects. The concentration of TNF-α and IL-1β of rat pleural effusion in GML, CMM and WMM group decreased significantly. The concentration of TNF-α, IL-1β and IL-8 in GML group, TNF-α, IL-8 in WMM group and IL-8 in CMM in rats serum decreased significantly. The body temperature in rats decreased significantly in GML and WMM group after 4-8 h of administration. CMM group showed no significant difference in rat body temperature compare with control. Compared with control group, GML (55-13.75 g•L⁻¹) could inhibit KPN biofilm formation and reduce number of viable cells in the KPN biofilm. CMM (45-22.5 g•L⁻¹) and WMM (10 g•L⁻¹) could also inhibit KPN biofilm formation and reduce number of viable cells (P<0.01). Result of SEM also showed that GML (55 g•L⁻¹) and its CMM (45 g•L⁻¹) and WMM (10 g•L⁻¹) could interfere the bacterial arrangement of KPN biofilm and extracellular matrix. GML and its CMM & WMM could inhibit the formation of KPN biofilm, CMM & WMM in GML showed synergism and complementation in inhibit KPN biofilm. Results showed that GML had obvious anti-inflammatory and antipyretic effects and could destruct KPN mature biofilm. WMM and CMM showed obvious synergistic effect against inflammation and inhibition of KPN biofilm formation and reduction of number of viable cells but no same effects against fever.

Study on toxicity of 999 Ganmaoling grain and influence of diet on hepatic toxicity / 中国中药杂志

This paper was aimed to compare the acute toxicity of 999 Ganmaoling grain and its different ingredients, and investigate the influence of routine diet on the hepatic toxicity induced by Ganmaoling in mice, so as to provide experimental basis for the clinical safety evaluation. Mice were given a single dose of Ganmaoling grain or its different ingredients respectively by gavage, and then observed for 14 days. LD₅₀ values of Ganmaoling grain or its chemical ingredient and the maximal tolerated dose of its herb ingredient were determined. Mice were divided into starvation and diet group, a single dose of Ganmaoling grain was administered by gavage. LD₅₀ values were estimated after 14 day observation. Mice were divided into starvation and diet group. At the same time,control group was set up for each. A single dose of Ganmaoling grain was given. Serum biochemical indexes were detected, liver weight index was calculated and liver tissue morphological change was observed after 6 h. LD₅₀ values were 4.42, 0.64 g•kg⁻¹ for Ganmaoling grain group and chemical ingredient group, respectively. The maximal tolerated dose of the herb ingredient group was close to 24.24 g•kg⁻¹. The toxic symptom was basically similar in the Ganmaoling grain and the chemical ingredient group. The body weight and food intake were decreased to a certain extent in both groups. There were pathological changes of liver and heart tissue in some of the surviving animals. The animals in the Ganmaoling grain group exhibited a lighter toxicity and recovered faster than that in the chemical ingredient group. LD₅₀ values of Ganmaoling grain were 2.56, 6.93 g•kg⁻¹ for starvation and diet group respectively. TD₅₀ values were 1.29, 6.31 g•kg⁻¹ for starvation and diet group respectively. The toxicity of 999 Ganmaoling was less, which may be related to the reduction of toxicity after the combination of herb and chemical ingredients. Compared with starvation group, the values of LD₅₀ and TD₅₀ of diet group was significantly increased, and toxicity was decreased. From the point of view of safety, it is safer to use Ganmaoling in the absence of hunger or after meal. The above tests provide experimental basis for the clinical safety use of Ganmaoling.

Tissue distribution of PEGylated puerarin in acute myocardial ischemia mode rats / 药学学报

The aim of this study is to explore the tissue distribution of PEGylated puerarin in acute myocardial ischemia model rats. Healthy male SD rats were randomly divided into two groups (30 each). Both were given PEGylated puerarin at a dose of 488 mg x kg(-1). After 5 min of medication, one group was normal rats, another group with acute myocardial ischemia was established by peritoneal injection of 50 mg x kg(-1) isoprenaline. After administration, the animals were executed at 30, 60, 90, 120, 150 and 180 min, then heart, liver, spleen, lung, kidney were extracted. The content of puerarin in organ tissue was determined by HPLC. The results showed that the AUC of tissue distribution of PEGylated puerarin in normal rats was liver > kidney > heart ≈ spleen > lung > brain. While the AUC of tissue distribution of PEGylated puerarin in acute myocardial ischemia model rats was liver ≈ heart > kidney > lung ≈ spleen > brain. AUC(heart) of PEGylated puerarin in acute myocardial ischemia model rats was 1.7 times than that of the normal rats, and there was significant difference (P < 0.05). Thus, PEGylated puerarin had a good heart-targeting property in early myocardial infarction area, drugs could accumulate in the ischemic myocardium. It provided important information for further study and clinic use of PEGylated puerarin.

"Re-evaluation upon suspected event" is an approach for post-marketing clinical study: lessons from adverse drug events related to Bupleuri Radix preparations / 中国中药杂志

We revisited the "Xiao Chaihu Decoction event (XCHDE)" occurred in late 1980s in Japan and the Bupleuri Radix related adverse drug reaction (ADR) reports in China After careful review, comparison, analysis and evaluation, we think the interstitial pneumonitis, drug induced Liver injury (DILI) and other severe adverse drug envents (ADEs) including death happened in Japan is probably results from multiple factors, including combinatory use of XCHDE with interferon, Kampo usage under modern medicine theory guidance, and use of XCHD on the basis of disease diagnosis instead of traditional Chinese syndrome complex differentiation. There are less ADE case reports related to XCHD preparation in China compared to Japan, mostly manifest with hypersensitivity responses of skin and perfuse perspiration. The symptoms of Radix Bupleuri injection related ADEs mainly manifest hypersensitivity-like response, 2 cases of intravenous infusion instead of intramuscular injection developed hypokalemia and renal failure. One case died from severe hypersensitivity shock. In Chinese literatures, there is no report of the interstitial pneumonitis and DILI associated with XCHDG in Japan. So far, there is no voluntary monitoring data and large sample clinical research data available. The author elaborated the classification of "reevaluation" and clarified "re-evaluation upon events" included the reaction to the suspected safety and efficacy events. Based on the current status of the clinical research on the Radix Bupleuri preparations, the author points out that post-marketing "re-evaluation upon suspected event" is not only a necessity of continuous evaluation of the safety, efficacy of drugs, it is also a necessity for providing objective clinical research data to share with the international and domestic drug administrations in the risk-benefit evaluation. It is also the unavoidable pathway to culture and push the excellent species and famous brands of TCM to the international market, in that it enhances the pharmaceutical manufactures the ability to control enterprise risk and eliminate the negative impact on traditional Chinese medicine. With the widely used and ADR report concentrated Radix Bupleuri injection as an example, the author provide a new approach to post-marketing clinical research.

Advances in studies on toxicity of aconite / 中国中药杂志

Aconite has the efficacy of reviving yang for resuscitation, dispelling cold and relieving pain, which is widely used in clinic, and shows unique efficacy in treating severe diseases. However, aconite has great toxicity, with obvious cardio-toxicity and neurotoxicity. Its toxicological mechanism main shows in the effect on voltage-dependent sodium channels, release of neurotransmitters and changes in receptors, promotion of lipid peroxidation and cell apoptosis in heart, liver and other tissues. Aconite works to reduce toxicity mainly through compatibility and processing. Besides traditional processing methods, many new modern processing techniques could also help achieve the objectives of detoxification and efficacy enhancement. In order to further develop the medicinal value of aconite and reduce its side effect in clinical application, this article gives comprehensive comments on aconite's toxicity characteristics, mechanism and detoxification methods on the basis of relevant reports for aconite's toxicity and the author's experimental studies.