Comparison of hepatotoxicity and toxic mechanisms of matrine and oxymatrine using in vivo and in vitro models / 中国比较医学杂志

Objective To compare the hepatotoxicity of matrine (MT) and oxymatrine (OMT) and explore the severity and characteristics of their toxicity, and to preliminarily elucidate their toxic mechanisms. Methods Liver cell line LO-2 cells were treated with acetaminophen (APAP), matrine and oxymatrine for 24 h, and the IC values, the contents of enzymes in the liver cells, the pathological changes, the contents of malondialdehyde (MDA) and glutathione (GSH) and the cell apoptosis rate were detected. In addition, adult zebrafish were treated with APAP, matrine and oxymatrine for 96 h, and the LC50 values, the pathological morphology of the liver cells, the contents of MDA and GSH and the apoptosis rate were detected. Meanwhile, the expression of oxidative stress-related gene, zgc: 136383, and the apoptosis-related genes, EIF4EBP3 and zgc: 123120, was also detected. Results Matrine and oxymatrine had toxic effects on liver cells in vitro. The IC50 value of matrine was 5. 3 mmol/L, and that of oxymatrine was ＞ 19 mmol/L. The contents of alanine aminotransferase (ALT), alkaline phosphatase (ALP), aspartate aminotransferase (AST) and lactate dehydrogenase (LDH) in the liver cells treated with matrine or oxymatrine were increased, and the cells appeared swollen, with an increase in the MDA level and a decrease in the GSH level. The cell apoptosis rate was also increased (P ＜ 0. 05). Furthermore, matrine and oxymatrine had toxic effects on the zebrafish. The LC50 value of matrine was 0. 41 mmol/L, and that of oxymatrine was ＞3. 8 mmol/L. The hepatocytes of zebrafish treated with matrine and oxymatrine appeared vacuolization in a mild to moderate degree, with an increase of the MDA content and a decrease of the GSH content. The cell apoptosis rate was increased (P ＜0. 05 for all). Expression of the oxidative stress-related gene zgc: 136383 (P ＜ 0. 05) and the apoptosis-resistant gene EIF4EBP3 (P ＜ 0. 05) was down-regulated by matrine, but that of the apoptosis-promoting gene zgc: 123120 was up-regulated (P ＜ 0. 05). Conclusions Results of the experiments using liver cells in vitro are consistent with those using the in vivo zebrafish model. Matrine (MT) and oxymatrine (OMT) both have hepatotoxicity, with similar toxic characteristics, and the toxicity of matrine is greater than oxymatrine. The mechanism of their hepatotoxicity is related with oxidative stress and cell apoptosis. Matrine reduces lipid transportation and activates oxidative stress reactions through down-regulation of gene zgc: 136383. In addition, matrine induces apoptosis in the liver cells via up-regulation of the apoptosis-promoting gene zgc: 123120 and down-regulation of the apoptosis-resistant gene EIF4EBP3.

Comparison of liver toxicity of matrine and oxymatrine in m ice / 中国药理学与毒理学杂志

OBJECTIVE To compare the liver toxicity of matrine and oxymatrine ,and to explore their toxic mechanism. METHODS Thirty ICR mice were randomly divided into normal control ,matrine 200 mg · kg-1 and oxymatrine 200 mg · kg-1 groups,10 mice per group. After single ig administration of corresponding drugs or water, animal mortality was calculated at the 15th day. The content of glutamic-pyruvic transami?nase(GPT),glutamic-oxalacetic transamin(GOT),alkaline phosphatase(ALP)and lactate dehydroge?nase (LDH) in serum were detected. Histopathological changes of the liver were examined by HE stain. The content of superoxide dismutase(SOD)and glutathione(GSH)in liver homogenates were detected by ELISA. Hepatocyte apoptosis was detected by Tunel stain. RESULTS The mortality rate of mice in two groups was 80% and 0,respectively. GPT,GOT and ALP contents of dead mice in matrine group were significantly higher than that in normal control group(P0.05). The content of SOD and GSH of dead mice in matrine group was lower than that in control group(P<0.05,P<0.01). The content of GSH in oxymatrine group was also decreased(P<0.05). The apoptosis rate of liver cells in dead mice in matrine group was increased(P<0.05). CONCLUSION A large dose of matrine and oxymatrine can produce liver toxicity. At an equal dosage,the liver toxicity of matrine is significantly higher than that of oxymatrine. The toxic mechanism is related to oxidative stress and apoptosis.

Metabonomic study on early biomarkers of hepatic injury induced by ethanolic extract from Rhizoma Dioscoreae Bulbiferae in rats based on 1H-NMR / 中国药理学与毒理学杂志

OBJECTIVE Dynamics of serum and urine metabolites in hepatic injury rats induced by ethanolic extract from Rhizoma Dioscoreae Bulbiferae(RDB)was investigated by 1H-NMR-based metabo?nomic methods in order to discover early biomarkers of liver toxicity induced by RDB. METHODS Rats were ig adminisetred with RDB at a dose of 5 g·kg-1 for 28 d. Rats were sacrificed 3,7,14 and 28 d af?ter RDB administration,as well as after a recovery period,respectively. Blood was taken for routine bio?chemical analysis by an automatic biochemical analyzer. Liver/body mass indexes were calculated ,and liver pathological changes were observed with hematoxylin-eosin staining. Urine samples were collected before and 3,7,14 and 28 d after RDB administration,respectively,as well as after withdrawal. Metabo?nomic analysis was carried out for serum and urine samples. Principal component analysis and orthogonal partial least squares-discriminant analysis were used for screening and identifiying early biomarkers. RESULTS Compared with the control group,total bilirubin (TB) and total cholesterol (TC) values were increased in 3-28 d in RDB group(P<0.05,P<0.01). Total bile acid(TBA)was elevated in 7-28 d (P<0.05,P<0.01). TB,TC and TBA became normal after discontinuation with RDB. There was no significant difference between RBD-treated group and control group in the activity of glutamic-pyruvic transaminase and glutamic-oxaloacetic transaminase,and the content of glucose also was not different between the two groups. The ratio of liver/body mass was elevated at 3-28 d(P<0.01)but returned to normal after withdraval of RDB. The enlargement and necrosis of hepatocytes were observed 7 d after RDB administration,and lesion degree was aggravated with the extension of RDB delivery time. Meta?bonomic analysis showed that the serum lipids (low density lipoprotein/very low density lipoprotein (LDL/VLDL),glutamic acid,choline phosphate and glycerolphosphatecholine were increased in the early stage. Pyruvate and N-acetylglutamate were decreased in urine. These metabolites became normal 7 d after discontinuation with RDB. CONCLUSION The serum lipids (LDL/VLDL),glutamic acid,glycerol phosphate choline,as well as urine pyruvic acid salt and N-acetyl glutamate may be used as the early biomarkers for liver toxicity induced by RDB.

Studies on Anaphylactoid Reactions Induced by Traditional Chinese Medicine Injections of Qingkailing and Xuesaitong / 中国中医药信息杂志

Objective To study anaphylactoid reactions induced by traditional Chinese medicine injections (TCMI) of Qingkailing (QKL) and Xuesaitong (XST) with RBL-2H3 cells;To provide some reference for improving the screening system of TCMI induced anaphylactoid reactions.Methods The IC50 induced by QKL and XST injections was determined by MTT assay. RBL-2H3 cells were stimulated with TCMI at different concentrations or with C48/80 or culture medium. Thirty minutes later, the supernatant was collected to determine the release of histamine andβ-hexosaminidase. The cell degranulation rate and the ultrastructure changes were observed. The ICR mice were given single injection of TCMI containing Evans Blue through tail vein. The number of the animal with blue ear, the total number of blue ears and the quantity of Evans Blue of extravasation were determined 30 minutes later.Results The IC50 of both QKL injection and XST injection was 12.5μL/mL. These two injections promoted RBL-2H3 cells to release histamine andβ-hexosaminidase at higher concentrations (P<0.05,P<0.01) in a dose dependent manner. Cell morphology showed a decrease of villous on the cell surface and an increase of the internal vacuolated structure. Both injections caused the blue ears of all animal with a rate of 100%. The quantity of Evans Blue of the extravasation was significantly increased (P<0.01). The results in vitro study were in close agreement with that in vivo.Conclusion Both QKL injection and XST injection may potentially cause anaphylactoid reactions. The RBL-2H3 cell model may be valuable to evaluate the anaphylactoid reactions induced by TCMI.

Advances of utilizing microRNAs as biomarkers / 中国中药杂志

MicroRNAs (miRNAs) are a new class of endogenous, single-strand, noncoding small RNAs. MiRNAs play an important regulatory role in a variety of pathological and physiological process, such as cell proliferation and apoptosis, organ development and differentiation and tumorigenesis and so on. It has been found that circulating miRNAs are also stably and specially expressed in serum or plasma and other body fluids. Circulating miRNAs could be taken as noninvasive and new biomarkers for evaluating the drug-induced target organ injury, which may play a vital role in monitoring the drug toxicity at the early stage.

Effects of external use of jiuyi dan for one month on blood and urine mercury levels and liver and kidney functions of rabbits / 中国中药杂志

<p><b>OBJECTIVE</b>To observe the changes of the blood and urine mercury (Hg) levels and liver & kidney functions of rabbits after administration of Jiuyi Dan (calcined gypsum-Sheng Dan 9: 1) for 1 month and the recovery of rabbits after the drug withdrawal.</p><p><b>METHOD</b>The rabbits were randomly divided into 2 groups: the calcined gypsum group and the Jiuyi Dan group. After 36 mg of calcined gypsum and 40 mg of Jiuyi Dan were used on the surface of wound (5 cm x 5 cm) on one side of rabbit back for 4 h, the surfaces of wound were washed by saline. The bloods were taken from the rabbit hearts before and after the drug administration for 14 and 28 days, and after the drug withdrawal for 7, 40, 71, and 92 days for determining Hg level in blood, and liver & kidney function indicators (ALT, AST, CREAT and BUN). The Hg level in urine collected from bladders was examined while rabbits were dissected after the drug withdrawal for 1, 40, 71, and 92 days.</p><p><b>RESULT</b>The Hg level in blood was significantly increased (P < 0.01) after the rabbits were administrated with drugs for 14 and 28 days and after the drug treatment was stopped for 7 and 40 days. The Hg level in urine was significantly enhanced after the drug withdrawal for 1, 40, 71 days. However, the liver & kidney indicators were not influenced.</p><p><b>CONCLUSION</b>The Hg level in rabbit blood and urine was significantly increased after the consecutive administration of double-dose Jiuyi Dan for 1 month. However, the blood Hg level and urine Hg level recover after the drug withdrawal for 71 days and 3 months, respectively. The liver & kidney indicators do not significantly change with the dose.</p>

Study on different doses of mercury-containing preparations on acute toxicity in rabbits / 中国中药杂志

<p><b>OBJECTIVE</b>To observe the effect of single administration of mercury- containing preparation Jiuyi Dan (calcined gypsum-Shengdan 9: 1) and Shengdan on acute toxicity of rabbits, in order to assess the safety of tested drugs.</p><p><b>METHOD</b>The rabbits were randomly divided into 4 groups: the calcined gypsum group (excipient control), the Jiuyi Dan group, the 90 mg Shengdan group and the 180 mg Shengdan group. After 270 mg of calcined gypsum, 300 mg of Jiuyi Dan, 90 mg of Shengdan, and 180 mg of Shengdan were used on the surface of wounds (5 cm x 5 cm) on two sides of rabbit back for 5 h, the surfaces of wound were washed by water. The bloods were taken from the rabbit hearts before and after the drug administration for 24 h, 72 h, 7 d and 14 d for determining Hg level in blood and liver & kidney function indicators (ALT, AST, CREAT, and BUN). The rabbits were dissected after the drugs treatment for 14 d, and pathological tests were made for their livers and kidneys.</p><p><b>RESULT</b>Compared with the calcined gypsum group, the 90 mg Shengdan group and the 180 mg Shengdan group showed significant increase (P < 0.01 or P < 0.05), as evidenced by increase in CREAT for 24 h and 72 h and increase in BUN for 24 h and on 7 d. AST is significantly increased as well (P < 0.01) for 24 h and 72 h compared to that of the group before drug treatment. The Hg level in blood was significantly enhanced (P < 0.01) after the rabbits were administrated with drugs for 24 h to 72 h. The pathological changes in livers and kidneys of rabbits were observed in the two doses of Shengdan treatment groups.</p><p><b>CONCLUSION</b>The Hg blood levels were increased significantly in an obvious dose-effect relationship in all drugs treatment groups. Liver & kidney function indicators were influenced by Shengdan treatment to some extent. Meanwhile, pathological changes in rabbit livers and kidneys were also caused by Shengdan, while Jiuyi Dan has no significantly effect on livers and kidneys.</p>

Mechanism studies on hepatotoxicity of rats induced by Sophorae Tonkinensis Radix et Rhizoma in rat / 中国中药杂志

<p><b>OBJECTIVE</b>To study the hepatotoxicity mechanism of rats that induced by Sophorae Tonkinensis Radix et Rhizoma.</p><p><b>METHOD</b>The SD rats were randomly divided into the control and Sophorae Tonkinensis Radix et Rhizoma (RRST) group, given distilled water and RRST 10 g x kg(-1) separately by orally for 7 days, and RRST 20 g x kg(-1) for other days until the 26th day. Blood was drawn from abdominal aorta after the rats were anesthetized by 25% urethane, and then centrifugally processed to get the serum for detection of ALT and AST. The liver tissues of control and experimental group were taken to prepare liver homogenate with cold NS, and centrifugally processed to get the supernatant. The activities of SOD and GSH, the content of gamma-GT and MDA were detected according to the methods of kit. The tumor necosis factor(TNF-alpha) was detected by ABC-ELISA. The expression of ICAM-1 was detected by immunohistochemistry.</p><p><b>RESULT</b>After the rats were given RRST by oral for 26 days, the ALT and AST activities in serum increased, the content of GSH and the ratio of SOD and MDA decreased, the content of gamma-GT and TNF-alpha, the masculine expression of ICAM-1 increased.</p><p><b>CONCLUSION</b>After the rats were given RRST, the liver can be damaged obviously, and the mechanism of hepatotoxicity perhaps related to free radical and inflammatory factor.</p>

An experimental serum pharmacological study on an application method in traditional Chinese medicine for treatment and prevention of asthma / 中西医结合学报

OBJECTIVE: To study the mechanism of herbal application along meridians for treatment and prevention of asthma by using serum pharmacological test to observe the effects of serum containing herbs against the constriction of tracheal spiral strips induced by acetylcholine chloride (Ach). METHODS: Guinea pigs were randomly divided into normal control group, normal saline (NS) application group, aminophylline application group, aminophylline injection group, 1-day herb application group, 7-day herb application group and 14-day herb application group. Asthma was induced by Hutson's method in guinea pigs except the normal control group. Guinea pigs in herb application groups were treated by external application of a compound herbal medicine 60 min once every day. Guinea pigs in NS application group were treated by external application of NS. Guinea pigs in the two aminophylline-treated groups were treated by external application and intraperitoneal injection of aminophylline at a dose of 400 mg/kg, respectively. The guinea pigs were killed and the sera were obtained after 1-day, 7-day and 14-day treatment in the three herb application groups, 7-day treatment in the NS application group, the aminophylline application and injection groups, respectively. Serum pharmacological method was used to do the experiment, the effects of different sera on the constriction of tracheal strips were observed, and the constriction rates were calculated. RESULT: The serum containing herbs had an effect in reducing the constriction of tracheal spiral strips induced by Ach, and the effect was similar to that of the serum obtained from the aminophylline injection group. The constriction rate of the tracheal spiral strips was decreased when herbal application treatment was prolonged within a period of time, and it became stable when herbal application treatment was between 7-14 days. The constriction of tracheal spiral strips induced by Ach could be reduced remarkably when it was previously treated by serum containing herbs. CONCLUSION: The anti-acetylcholine function with a time-dependent effect is one of the mechanisms of herbal application treatment along meridians for asthma, and furthermore, herbal application treatment along meridians might be useful for preventing asthma.

Studies on liver-protection and promoting bile secretion of gentiopicroside / 中草药

Object To study the liver-protection and bile promoting secretion of gentiopicroside administrated ig in rat. Methods Gentiopicroside was administrated ig to the mice with liver injury induced by CCl 4 and D-GlaN. ALT, AST levels in serum and GSH-Px activity in liver were examined. The bile flow rate and the concentration of the main components in the bile of rats after ig administration of gentiopicroside were estimated. Results Gentiopicroside administrated ig decreased the serum ALT and AST levels, increased the liver GSH-Px activity in the mice treated with CCl 4, promoted the secretion of bile and increased the concentration of bilirubin in the bile. Conclusion Gentiopicroside administrated ig exhibited significant liver-protection and promoting bile secretion.

Experimental Study of Gout Granule f or Gout / 中药新药与临床药理

Objective To investigate the effect of Gout Gra nule(GG)for gout.Methods Gout -relieving effect of GG was observed in rats with adjuvant arthritis induced by sodium urate and i n mice with hyperuricemia induced by hypoxan-thine.Anti -inflammation of GG was i nvestigated in mice with xylene -ind uced auricular swelling and in rats with edema of hind paw induced by injection of carrageenin.WBC migratory response c aused by carboxymethylcellulose(CMC)in mice was also observed.Results GG could reduce the blood level of uric acid and inhibit acute inflammatio n in model ani-mals and also tended to promote the elimination of uric acid.Conclusion GG had a remarkable effect on the acut e attack of gout.

Study of Anti-infection Effect on Jindengshangen Oral liquor / 中成药

Objective: To study the mechanism of jindengshanggen (JDSG) oral liquid's anti infectiion effect.Methods: inhibition on bacteria growth, resistance to Escherichia coli endotoxin and measurement of monocytic phagocaryosis in mice were taken. Results: The effects of JDSG oral liquid is probably not due to the inhibition on bacteria growth, but rather due to its stimulation on other anti infection mechanisms in human body. The enhancement of immune response and toxin tolerance may be one of the mechanisms for JDSG oral liquid to cure acute pharynx infection. Conclusions: These results provide a basis for clinical application of JDSG.

Preparation of Blood-Deficient Model and Effects of Tangkuei Blood-Supplementing Decoction on Hemopoiesis / 中成药

Objective: To prepare a blood deficient model and study the effect of Tangkuei Blood Supplementing Decoction on hemopoiesis in this model. Methods: This model was made in mice by i.p. an accumulate doses of 60mg/kg acetylphenylhydrazine (APH) and 160mg/kg cyclophosphamidum (CY). The RBCs, WBCs, reticulocytes and bone marrow nucleated cells (BMNC) were counted, the micro structure of bone marrow was observed. The swimming time, the body temperature and the plasma cAMP, cGMP levels were measured. The effects of Tangkuei Blood Supplementing Decoction by p.o. administration on promoting the hemopioetic function were observed with the model mice. Results: Tangkuei Blood Supplementing Decoction could remarkably increase RBC, WBC, BMNC, improve the proportion of reticulolytes in peripheral blood and the micro struture of bone marrow, prolony the swimming time, raise the body temperature and the specific value of cAMP/cGMP. Conclusion: The model exhibits the main features of blood deficiency, and can be used as one of models of blood deficiency. Tangkuei Blood Supplementing Decoction can obviously improve the dual deficiention of qi and blood of model mice by supplementing qi and blood.

Chromatography fingerprint of the constituents of Melia toosendan Sieb / 中成药

AIM:To establish the chromatography fingerprint of the constituents of Melia toosendan Sieb from different areas in order to provide a base for the identification of its quality. METHODS: A gradient separated method was applied. Column:Inertsil ODS-3 C_ 18 ,mobile phase:acetonitrile-water,detection wavelength:270 nm,flow rate:1.0 mL/min,column temperature:25 ℃. RESULTS: To establish the chromatography fingerprint of the constituents of Melia toosendan Sieb., make the technical parameters for its quality controlling,and mark 39 main peaks as its characteristic fingerprint. CONCLUSION: The distribution of constituents of Melia toosendan Sieb differ a little from different areas, but the propotion of the constituents differ greatly, with the Melia toosendan Sieb.from the same area , the distribution and propotion differ a little. This method is reproducible, simple and easy, and can be use to provide a base for the quality control of Melia toosendan Sieb.

Effect of peppermint oil on GSH and ATPase in rat liver-tissues and primary cultured rat hepatocyte / 中成药

AIM: To study the effect of peppermint oil on GSH、ATPase in rat liver-tissues and primary cultured rat hepatocyte. METHODS: The rat liver tissues were obtained to detect content of GSH and level of Na~+-K~+-ATPase、Ca~(2+)-Mg~(2+)-ATPase after being administered peppermint oil orally at 24 % concentration of peppermint oil in 36 and 48 hours.Primary rat hepatocyte was separated and cultured,then peppermint oil and the serum containing that were respectively added level of LDH,ALT,AST in the supernatant fluid was respectively determined after incubating for 12,24 and 48 hours. RESULTS: The content of GSH and level of Na~+-K~+-ATPase and Ca~(2+)-Mg~(2+)ATPase in liver tissues was low remarkably(P

The effect of Gengnianle Granule on hypothalamic-pituitary-ovarian axis in climacteric rats / 中成药

AIM: To investigate the effect of Gengnianle Granule on hypothalamic-pituitary-ovarian(HPO) axis in climacteric rats.METHODS: 12-15 months SD female rats were randomized into climacteric group and young control group was selected additionally.After being administrated for thirty days,the level of E_2、P、Te、FSH、LH in serum were examined by the radio-immunity method,the expression of GnRH in hypothalame,the expression of ER in hypothalame and pituitary appendage were examined by the immunohistochemical method. RESULTS: Compared with climacteric group,Gengnianle Granule could increase the level of E_2、P in serum(P

Study of Xiaoyao Tablet on hepatoprotective action / 中成药

Objective: To study the protective effect of Xiaoyao Tablet(XYT)(Radix Bupleuri, Radix Angelicae Sinensis, Rhizoma Atractylodis Macrocephalae, Radix Paeoniae Alba, Poria, Radix Glycyrrhizae, Herba Menthae, Roasted ginger) on liver. Methods: Using the acute hepatic injury model induced by Tetrachloride and D-galactosamine (CD-GaIN) in rats or mice, we observed the influence of XYT on ALT and AST level in the serum, MDA level and glutathione-s-transferase(GST) activity in the liver homogenates. The pain-killing effect was tested by body twisting model induced by glacial acetic acid and by hot plate model in mice. Results: The XYT reduced the serum ALT level significantly in the acute hepatic injury model in rats and mice. In the CCl 4 reduced acute hepatic injury mice model, XYT decreased the MDA level and increased the GST activity in liver significantly, also it reduced the MDA level in the serum. The XYT reduced twisting number in mice induced by glacial acetic acid. In the hot plate test, the pain threshold was increased. Conclusion: The drug has the similar hepatoprotective function to Xiaoyao Pills.

Activation of Na~+/H~+ exchange 1 and myocardial hypertrophy / 中国药理学通报

Na +/H + exchanger 1 (NHE1) is a major contributor to ischemic and reperfusion injury. It is emerging that NHE 1 also contributes to myocardial hypertrophy and heart failure due to chronic maladaptive stimulation. It appears that NHE 1 may represent a common downstream mediator for various hypertrophic factor, such as angiotensinⅡ,beta (1) and alpha (1) adrenergic receptor activation. NHE 1 inhibition may be a effective new therapeutic approach for prevention and treatment of heart failure.-

Protection of the ischemic and reperfused myocardium by sodium-hydrogen exchange inhibitor / 中国药理学通报

During ischemia, the intracellular acidosis stimulates the Na +/H + exchanger. The increase in intracellular sodium secondary to increased Na +/H + exchange is reversed through exchange for calcium, resulting in calcium overload of the cells and producing cell injury and necrosis. NHE1 inhibitors exert a protective effect on myocardium subject to ischemia and reperfusion via reduction of Na +/H + exchange, attenuation of intracellular calcium accumulation and other mechanisms.