Gleditsia Sinensis Extract On Liver Cancer Cells In Mice TGF-β/Smads Regulation Of Signal System

Objective:About 100 million people worldwide each year die from liver cancer. In recent years, studies of tumor cell signal transduction pathway into the development of mechanisms and clarify the tumor biology to find an effective treatment for hot spots. The results show that abnormal signal transduction TGF-β/Smads tumor invasion and metastasis is an important factor. TCM pathogenesis of tumor formation is complex and that the phlegm resistance is the key to the pathogenesis of tumor formation. Zhu Dan-famous medical scientist in his major works are listed with sputum door of their hair cut herbs, especially in tumors raised awareness of risk factors related with sputum. Acacia is a "phlegm dampness, Sanjiexiaozhong" anti-tumor efficacy of drugs commonly used. We hope that by studying the acacia extract (ethyl acetate extract) on liver cancer cell signaling system in the regulation of TGF-β/Smads for acacia extract (ethyl acetate extract) it has a reference against liver cancer.Methods:PartⅠ:Theory1 From the Chinese medicine theory to clarify the relationship between sputum and tumor. Stressed sputum, drugs and stasis interaction, mutual transformation, tumorigenesis and mutual stalemate is key to development. 2 raised from the "phlegm" On the feasibility of liver cancer treatment. And set out phlegm medicine "acacia" anti-cancer based on a preliminary study.PartⅡ:the literature of acaciaFrom the general properties of acacia; acacia historical study; with the prescription of acacia; acacia pharmacological research and clinical applications such as acacia acacia clarify the characteristics and research value.PartⅢ:Experimental StudyWe use the Kunming mice of clean grade 60. Through the random method (random number table) to experimental animals were divided into 6 groups, were divided into negative control group (Tween 80 solvent); honey locust extract (ethyl acetate extraction) 3 concentrations (26.0mg/d,13.0mg/d,7.8mg/d); positive control group (Jinlong Capsule group); blank control group (not in the model).10 mice in each group. Modeling using the right mouse inoculated subcutaneously with H22 liver cell suspension method. 2 days after modeling, to 16 days, the two experimental groups and requirements (amount of each administered 0.4ml, already with a good concentration) were administered.17 days after modeling, the mice were killed to collect samples of the inspection. Experiment 1: the general situation of mice; mortality of mice in each group; stripping of mice tumor weight, tumor inhibition rate of each group and Pathology. Experiment 2:detection of serum levels ofγ-glutamyl transferase (GGT), alkaline phosphatase (ALP), total protein (TP), albumin (ALB) and alpha-fetoprotein (AFP) concentration. Experiment 3:Real-time fluorescence quantitative method to detect the liver tissue of mice and Smad7mRNA Smad4mRNA expression. Experiment 4:Immunohistochemistry was used to detect the expression of Smad4 and Smad7 protein.Results:Experiment 1:acacia extract inoculated mice bearing H22 hepatoma cell biological characteristics of normal and pathological observations1 observation:control activities in the normal mice, in good spirits, normal hair, and shiny, eating normal, urine normal. Acacia extract (high dose group), acacia extract (middle dose group), acacia extract (low dose) and the positive control group (dragon capsule), hair loss in mice in each group, there are sparse feel, gloss poor, listlessness, decreased activity, diet significantly reduced compared with the control group, less drinking water than the control group decreased obvious, toilet cases less than the control group, the outside world is slow to respond. Negative control (solvent soil temperature of 80) will be similar. Right flank of the mice inoculated with H22 hepatoma cells, the inoculation site lumps of varying sizes. Some parts of mouse peritoneal swollen abdomen, palpation with the fingertips, there is obviously a sense of volatility, consider the formation of ascites.2 Kunming 21.67% mortality in mice.3 Comparison of tumor weight of mice in each group:acacia extract (high) the average tumor weight of 2.87±0.34, acacia extract (in) the average tumor weight of 3.68±0.34, acacia extract (lower) the average tumor weight of 3.64±0.36, positive control group The average tumor weight of 2.60±0.36 and the average tumor weight of negative control group decreased significantly compared to 4.73±0.40, P<0.01; the negative control group, acacia extract (middle), acacia extract (low) and the positive control group was significantly higher, P<0.01; acacia extract (high) and the positive control group no significant difference in tumor weight, P>0.05.The mice tumor inhibitory rate:45% positive control; negative control group was 0; acacia extract high-dose group 39%; acacia extracts dose group 22%; acacia extract low-dose group was 23%.4 Pathological observation:the control group's eye and liver under the microscope all appear normal, model group mice in each group overall than the control group (non-model group) have more abnormal performance.Negative control group, acacia extracts dose group and low dose of acacia extract pathological observation:visual general observation:the liver volume than the control group mice increased significantly; texture than the control group increased hardness; liver surface shows multiple nodules scattered in the gray. The most negative group, acacia extracts dose group and low dose group of acacia extract the control group slightly less than the negative. Optical microscope:hepatic lobules was not clear, liver cells arranged in irregular claims. Obviously abnormal nuclear shape, some nuclei of liver cells increases, or increases the proportion of nuclear transfer.Positive control group, high dose of acacia extract pathological observation:general observation of the naked eye:the liver volume than the control group mice increased slightly; texture and hardness than the control group slightly increased, but less than the negative control group; liver surface can be scattered in the gray nodules occasionally. Positive control group and the acacia extract high-dose group was not significantly different. Optical microscope:hepatic lobules unclear, the liver cells arranged in irregular claims. Abnormal nuclear morphology was not obvious. Speculated that the high dose group of acacia extract through the inhibition of liver cancer tumors are significant growth of cancer cells, liver cells and thus delay further liver damage caused to serve the purpose of protecting the liver.Experiment 2:inoculation of acacia extract on mice bearing H22 liver cancer cells related to indicators of liver functionlevels of serum GGT:the positive control group average of 23±2 U/L, acacia extract group (high) mean 23±2 U/L and the negative control group averaged 38±2U/L compared significantly lower, P<0.01; acacia extract group (middle) mean 27±2U/L, acacia extract group (low) average of 37±2U/L compared with the negative control group average lower P<0.05. Acacia extract group (middle), on average, acacia extract group (low), on average, the average negative control group compared with the positive control group mean decreased, P<0.01.Serum ALP level:the positive control group mean 25±5 IU/ L, acacia extract group (high) mean 34±6 IU/L, acacia extract group (middle), average 56±5 IU/L and negative control group, mean 90±8 IU/L were significantly decreased, P<0.01; acacia extract group (low) average of 75+7 levels lower in the negative control group P<0.05. Acacia extract group (high), acacia extract group (middle), acacia extract group (low), ALP level in negative control group compared with the control group decreased significantly, P<0.01.Serum TP:positive control group average of 70±2, acacia extract group (high) mean 65±6, acacia extract group (middle), mean 59±3 and acacia extract group (low) average of 58±3 of the TP level in the negative control group average of 47±4 were significantly increased, P<0.01;. Acacia extract group (middle), acacia extract group (low) and the negative control group TP level in the positive control group decreased significantly, P<0.01.Serum ALB:positive control group mean 30±2, acacia extract groups (high) mean 29±1 in the mouse ALB level in the negative control group average of 22±2 were significantly increased, P <0.01. Acacia extract group (middle), mean 25±3, acacia extract group (low) average of 23±2 and the negative control group of mice ALB level in the positive control group decreased significantly, P<0.01.Serum AFP:Results mice AFP values were not measured.Experiment 3:acacia extract inoculated mice bearing H22 hepatoma cells TGF-β/Smads signal transduction pathway in smad4 mRNA and smad7 mRNA expressionSmad4mRNA expression:positive amplification control group mean 0.566±0.092 and acacia extract high-dose group averaged 0.471±0.069 and the average negative control group was significantly higher compared 0.320±0.099, P<0.01; acacia extracts dose group was Value of 0.363±0.058 and acacia extract low-dose group averaged 0.280±0.067 compared with positive control group decreased significantly, P<0.01. Smad7mRNA expression:positive amplification control group mean 2.007±0.499, acacia extract group (high dose) mean 2.230±0.491 in the mouse liver tissue Smad7mRNA expression level in the negative control group average of 3.537±0.401 significantly lower compared, P<0.01.Acacia extract group (middle dose group) mean 3.106±0.410, acacia extract group (low dose group) average of 3.613±0.416 and the negative control group of mice the expression of liver tissue Smad7mRNA compared with the control group significantly increased, P<0.01.Experiment 4:acacia extract inoculated mice bearing H22 liver cancer cell signal transduction pathway TGF-β/Smads smad4 and smad7 protein expressionSmad4 protein expression:positive control group of mice liver protein levels of Smad4 average 2973.190±248.540 and the average of the negative control group was significantly higher compared 1196.399±837, P<0.01. Acacia extract group (high dose) average 2295.187±820.119 increased compared with the negative control group, P<0.05.Negative control group of mice liver Smad4 protein levels compared with the positive control group decreased significantly, P<0.01. Acacia extract group (middle dose group) average 1558.835±302.801 and acacia extract group (low dose) average 988.614±338.995 decreased compared with the positive control group, P <0.05.Smad7 protein expression:positive control group average of 1454.381±302.405 and acacia extract group (high dose group) average 1494.202±399.553 Smad7 protein expression in mouse liver, the level of the average negative control group was decreased significantly compared 2239.589±272.892, P< 0.01. Acacia extract group (middle dose group) average 1496.981±505.121 decreased compared with the negative control group, P<0.05. Negative control group and acacia extract group (low dose) average 2330.056±177.105 in the mouse liver Smad7 protein levels compared with the positive control group was significantly higher, P<0.01.Conclusion:The acacia extract (ethyl acetate extract) of anti-liver cancer cells may have mechanisms to achieve the following:①to improve the quality of life of mice and inhibit the tumor on the body damage.②inoculation site by inhibiting the size of the tumor play a role in cancer.③tumor-bearing mice by protecting liver function and play a role in protecting the liver④acacia extract the high dose group (26.0mg/d) through enhanced expression and reduced Smad7mRNA Smad4mRNA expression.⑤acacia extract high-dose group (26.0mg/d) through increased Smad4 protein expression and reduced Smad7 protein. Play a role in comprehensive cancer.