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15regulation, anticoagulation, conduction mitosis information, and promote cell secretion and other important biological functions. Early studies focused on annexins involved in the inflammatory mechanism in its response, in recent years, more and more studies have found that in the process of tumor development and may play an important role, such as mediated apoptosis and differentiation. And with annexin A1, annexin A2, annexin A5 related reports more, while traditional Chinese medicine for diabetes state regulation of annexin A4 no special report. Annexin A4 (anxa4), belonging to annexin family, whose gene is located on 2p13, the protein exists in various tissues and organs of animals and plants, accounting for the total amount of intracellular protein 2%. Which is a Ca2 +-dependent phospholipid binding protein, a phospholipase inhibitor, combined with calcium ions, and calcium-dependent phospholipid binding activity can promote membrane fusion, in cyst transport, a calcium ion channel-forming, cytoskeletal activity, phospholipid and regulation and function of membrane receptor endocytosis, exocytosis and secretion of other important biological functions [2]. Recently reported in the literature annexins A4 may regulate cell growth and differentiation, regulation of apoptosis and other biological functions [3], and these biological behavior disorders may be associated with diabetes incidence of myocardial injury has a strong correlation. Studies have shown that annexins A4 can inhibit the intracellular Ca2 + levels, the maintenance of intracellular Ca2 + levels of stability [4], thus avoiding myocardial contractile dysfunction, but also may increase the resistance of cells to apoptosis.
The experimental results show that annexins A4 gene chip in the diabetic state downregulated myocardial tissue, suggesting that diabetic cardiomyopathy may occur with annexins A4 gene regulation related to its mechanism may be that: (1) regulation of calcium imbalance caused by myocardial contractile dysfunction, cause or aggravate diabetic cardiomyopathy myocardial injury; (2) is not conducive to the formation of the skeleton structure of myocardial cells; (3) myocardial cell growth, differentiation or apoptosis causes abnormal myocardial injury. 5.Conclusions
Our results showed that diabetic cardiomyopathy annexins A4mRNA expression was low.However ,its expression increased after treatment indicating candy hearts. Concord diabetic rat heart by upregulating the expression of annexins A4mRNA protect cardiac tissue , delaying further development of the disease. References
[1] Gerke V, Moss SE.Annexins :From Structure to Function [J]. Physiol Rev.2002,82(2):331.
[2] Mattin I,Dominguez F,Avila S,etal. Human endometrial receptivity,gene regulation[J]. Reprod Immunol.2002,55:131-139.
[3]Piston P, Ferrari D, Rapizzi E,etal The Ca2+ concentration of the endoplasmic reticulum is a key determinant of ceramide-induced apoptosis:significance for the molecular mechanism of Bcl-2 action. [J].Embo .2001,20(11):2690-2701.
[4]OkuseK,Malik-HallM,Baker MD,etal.Annexin II light chainregulates sensory neuron-specific sodium channel expression. [J] Nature.2002,417(6889):653-656.
The Animal Model of Insulin Resistance Induced by High-fat Diet
SONG Ying-xing
(Heilongjiang University of Chinese Medicine,Harbin,150040,China)
Abstract: Insulin resistance (IR) is type 2 diabetes mellitus (T2DM), impaired glucose tolerance(IGT), hyperlipidemia, hyperuricemia, cardiovascular disease and metabolic syndrome common disease foundation. In lab studies, the establishment of animal model of insulin resistance is the primary condition, The resistance way about it can be divided into four kinds: genotype, drug injection type, diet induced type, various ways mixed. This article mainly discusses on the type high
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fat diet induced insulin resistance in animal experiments, different high fat feed formula ratio, laboratory animal age, food intake and animal strains on the influence of the model. Conclusion: fat calories than high, food intake, old animals are more likely to develop the insulin resistance model.
Keywords: High-fat Diet, Insulin Resistance (IR), Type 2 Diabetes Mellitus (T2DM), Animal Model
1 Background
Insulin resistance (IR) refers to a variety of reasons make insulin to promote glucose uptake and utilization efficiency decreased, the reactivity decreases insulin physiological function, i. e., impaired insulin to promote glucose uptake, compensatory insulin secretion grow in quantity, cause its important symbol for high insulin hematic disease [1]. Insulin resistance and type 2 diabetes, metabolic disorder is secondary low glucose tolerance, reduce, hyperlipidemia, high uric acid hematic disease, cardiovascular disease and the common basis as the onset of metabolic syndrome. Through experiment study of the disease state is a major way in this field of research. In all animal experiments, insulin resistance model is the foundation and the prerequisite of research.
Insulin resistance there are many kinds of animal model way, main ways can be divided into the genetic type, drug induced type, diet induced type [2-5]. Here we mainly talk about diet induced by high-fat diet induced in the model. This way is through pure rats fed high fat, high quantity of heat of feed, inducing the formation of rats with insulin resistance, for all kinds of the most commonly used in the study of the dies can be made.
Human induced insulin resistance there are many reasons, of which the unhealthy, high-calorie, high fat diet is one of the main risk factors [6]. High-fat diet induced rat insulin resistance mechanism is simulated human unhealthy high fat diet, more embodies the factors such as environment, bad life habits, role in the pathogenesis of insulin resistance. Research found that chronic high fat diet, high caloric intake easily induced obesity, affect the skeletal muscle cell membrane surface glucose transporter 4 (glucose from 4, GLUT4) protein expression, make insulin stimulates GLUT4 transposition to the cell membrane, affect the insulin signal transduction system, so as to promote the formation and development of insulin resistance [7]. And affect glucose stimulation of islet cell insulin secretion 4-6, and insulin resistance induced by high-fat diet in rats, the liver, adipose tissue of c-reactive protein (C - reactive protein, CRP) expression increased, while adiponectin expression of adipose tissue fall in [8]. Another study found that chronic high fat feeding rats exists endothelial dependent vasodilation function decreases, its blood free fatty acid (FFA), free fatty acid, and triglycerides (total triglyceride, TG) levels increased [9]. Reason induced by high fat feed experimental animals to establish animal model of insulin resistance, stable and reliable, simple and practical, its main performance for high blood sugar, high weight, high insulin hematic disease, etc. Characteristics.
2 The influencing factors of building
Causing a wide number of risk factors for insulin resistance, in the process of experiment, some changes will cause great change. Alone to explore a high-fat diet on rats with insulin resistance mechanism, through the experiment observation, analysis of its own. Comparison of similar experiments and other researchers found that the following factors is different, can cause the experimenter building successful time and the quality of the similarities and differences.
2.1 Feed the fat composition and amount of feed intake
High fat feed, as the name implies, its main difference formula with ordinary feed fat accounts for heat than different. Fat into saturated fat and monounsaturated fat, not similar source of fat, saturated fatty acids mostly exists in animal fat, healthy diet, fat accounts for heat ratio should be below 30%. Currently on the market fat rats a high-fat diet contains mostly from lard [10-12], there is also some safflower oil [13]. In theory, the higher the fat content of feed easier to induce rats of insulin resistance, and also because of the shorter the amount of time. But due to the high fat feed in adipose content is higher, the production, processing, and save for more difficult. And the higher the fat content in the feed, the texture is soft, the quality of a material does not conform to the rodents diet habits, so easy to influence food intake of the rats, decrease independent food intake in
rats, so as to affect the total calorie intake in rats, but unfavorable to induce insulin resistance. So although currently on the market have different high-fat forage varieties used in the building, but its fat calories in usually account for 40-65%, average feed fat calories often for 5-10%. the United States Research Diets company produces feed as an example, the feed formula is as follows (table 1)
Table 1: ordinary feed heat and high fat feed ratio_
High fat feed(D12451) WM$(D12450B)
Content(%) Energy ratio(%) Content(%) Energyratio(%)
Protein 24 20 19.2 20
Carbohydrates 41 35 67.3 70
Fat 24 45 4.3 10
The high fat feed fat calories than 45%, average feed only 10% fat calories. It is not hard to see, than ordinary fat content in the feed is much lower than the high fat feed.
Compared to a lot of experimental literature, we can see fat accounts for heat than different, and building the amount of time is not the same. Child actors such as [14] in fat calories than 38. 5% of the feed six weeks of male Wistar rats, feeding to 12 weeks, the two groups of indicators, there is no obvious difference, acquired a high fat feed with lard, reach 51. 3% of fat calories than, continue to feed after 4 weeks, weight, blood sugar data appeared difference, in the process of the experiment on an empty stomach blood sugar levels, fasting blood glucose, FBG) slight fluctuations, but was no significant difference between the two groups (P > 0.05). High fasting plasma insulin high-fat group than the control group 45. 7% (P < 0.01), and weight growth are significantly lower than 59% in fat calories than feeding rats [15]. Wu Yanli [16], such as calories as fat accounted for 56.0% of the high fat feed weight about 180 g, male Wistar rats with 5 ~ 6 weeks, 12 weeks can build associated with insulin resistance when the glucose tolerance of the animal model of low cut. By 12 weekends weight of model group was obviously higher than that of control group, insulin and HOMA IR index is significantly higher than the control group. The above experiment suggests that the proportion of the high fat, high heat affected by the induction time of rats with insulin resistance. Analysis found that levels of rat diet during the experiment, has some influence on insulin resistance in morning and evening [17], the principle of general and high fat feed used in the building of the high and low fat content. Restricted diet, less food intake in rats, the intake of total calories less. Conversely, independent diet, intake of big rats is their total calorie intake is big, so the module and the amount of time is short. ReimerM K etc. The researchers used as much as 58% fat content high fat feed, free food, experimental group in 8 weeks [18] the hyperinsulinemia. Domestic scholars use the same fat calories than feed, free food, feeding at 4 weeks FBG significantly increased (P < 0.05), 5 weeks FINS began to rise, but there was no significant difference, at 10 weeks triglycerides, total cholesterol, high-density lipoprotein cholesterol, low density lipoprotein cholesterol were higher (P < 0.01). HE staining showed moderate fatty liver. [19]. But under the same fat calories than the high fat feed, Gianluca Svegliati - Baroni foreign researchers, according to the saturated fatty acids and unsaturated fatty acids and rat body weight than column, restricted rats to eat each week, in the 3 months (12) weeks rats blood glucose, alanine aminotransferase, alanine aminotransferase, ALT) increased significantly. [20]. Also have domestic scholars in the contrast different fat content of feed's influence on the metabolic syndrome in rats, while the high-fat group rats (divided into two groups, the feed fat calories than 45% and 60%, respectively), the body weight, blood sugar than normal group (feed fat calories more than 10%) increased significantly. But is found within the group, with 60% fat calories than the fat content in the feed is high, the quality of a material soft, thus unfavorable to eat rats, affect the total calorie intake, so the blood sugar of rats eating this kind of feed, weight growth without fat calories instead of 45% rats increased significantly. When considering 45% of high fat feed for induced insulin resistance may be better than 60% of the high fat feed [21].
2.2 Building the initial weight and weeks in rats
Weeks of animal experiments in rats of insulin resistance is another influencing factor of the building, namely experimental rats used in initial weeks to induce insulin resistance index of time and also have an impact. At the same fat calories than feed, Xu Lingling [22], such as feeding 14 weeks of age, body weight at about 270-320 - g SD rats, rats fed 60 days test group appear obvious insulin resistance, the 5/8 of the rat blood sugar value is greater than 8.3 tendency/l, and a control group and no greater than this value. Leptin and the test group and the serum insulin levels are significantly higher than the control group. And feed such as BuShi 6, 8 weeks of SD rats fed on 16 weeks experimental group rats fasting plasma free fatty acid (fasting free fatty acid, FFFA) was significantly higher than the control group.
2.3 Strain
Insulin resistance model is commonly used for rats and mice, animals commonly used strains with Wistar rats, Sprague wrote Dawley (SD) rats and C57BL / 6 j mice, and in the majority with males. Wistar rat is relatively smooth and is now the most widely use the longest time, the scope of experiment with rats, widely used in all kinds of biological research trials. SD rats is more fierce and the growth development period is long, weight gain than Wistar rats quickly, has the good infectious disease resistance ability of the endocrine system developed, reason often used in endocrine class experiment, bai and other researchers in fat calories than for 66.4% of the high fat feed SD rats, 8 week ACC mRNA levels in rat skeletal muscle increased over time, but the serum insulin levels are normal diet group increase is not obvious. [26]. In addition, C57BL / 6 j mice with ob/ob genetic background, sensitive to fat induced high blood, reason often used in the study of metabolic diseases [27]. Li Kang < http://www.ncbi.nlm.nih.gov/pubmed/? Term = Kang L [auth] > 60% of researchers in high fat feed C57BL / 6 j mice 20 w which appear obvious high insulin hematic disease, models with insulin resistance in mice inflammation [28]. Therefore, in establishing the animal model of insulin resistance, should choose suitable experimental animal strains. Choose a different animal strains also has different effects on the result of the experiment.
3 Conclusion
In any animal testing, the test model is the first condition of experiment, and the key to success. In the method of established animal model of insulin resistance, drug injection of STZ method although rapid, practical, but the method of experimental animals islet function damage is bigger, the normal physiological function of the law against animals and dose not easy control [29]. Used and simple way, induced by a high-fat diet time reasonable, economical and practical, and with the human type 2 diabetes, insulin resistance induced by environment, the modeling way is better for combination of animal experiment and clinical application. Through contrast and analysis, we found that in pure animal model of insulin resistance induced by high-fat diet, during the process of the establishment of experimental animals, strains, food intake, age or even high fat feed composition ratio, etc all can affect the process. Compared with high fat feed fat than high, weeks of experimental animals, animals are more likely to induce the calories more insulin resistance. Experimental animals, in Wistar rats and SD rats and C57BL / 6 j mice are commonly used to build model animals. Animal insulin resistance model is affected by many factors, the literature reports the amount of of all kinds of experiments made mold time and indicators for the quality of the similarities and differences can be affected by the above factors.
References
[1]Li Xiujun. The insulin resistance syndrome [M]. Beijing: People's Medical Publishing House, 2001.44-57.
[2]Oana Sandu,Song K, Cai W.et al.Insulin Resistance and Type 2 Diabetes in High-Fat-Fed Mice Are Linked to High Glycotoxin Intake[J].Diabetes J 2005 Aug;54(8):2314-9.
[3]Liang JL, Feng ZK, Liu XY,et al.Effect of impaired glucose tolerance on cardiac dysfunction in a rat model of prediabetes[J].Chin Med J(Engl).2011 Mar;124(5):734-9.
[4]N.Harishankar,Vajreswari A, Giridharan NV.WNIN/GR-Ob-An insulin-resistant obese rat model from inbred WNIN strain[J].Indian J Med Res.2011 September;134(3):320-329.
[5]Bandaru P,Rajkumar H, Nappanveettil G.Altered or Impaired Immune Response to Hepatitis B Vaccine in WNIN/GR-Ob Rat:An Obese Rat Model with Impaired Glucose Tolerance.ISRN Endocrinol[J].2011;2011:980105.
[6]Ershow A.Environmental influences on development of type 2 diabetes and obesity:challenges in personalizing prevention and management.Journal of diabetes science and technology[J].2009 Jul 1;3(4):727-34
[7] Song Chunyu, bi will people. High fat diet feeding effect on rat skeletal muscle cell membrane GLUT4 content [J]. Chinese journal of pathology, physiology, 2004, 20 (10) : 1866-1866.
[8] Yuan Guoyue, Yang ling, jia, etc. High fat feeding fat tissue of the rats induced insulin resistance c-reactive protein and the expression of adiponectin change [C]. Beijing; 5 th national conference on endocrine metabolic disease with traditional Chinese and western academic BBS and diabetes, 2012.
[9] Yang Huangguang. Long-term high fat feeding on SD rat aortic endothelial dependent vasodilation function on the effects and mechanism of research [D]. Beijing, Beijing union medical college, 2006.
.[11]Reimer M K,Ahre'n B,et al.Alteredp-cell distribution of pdx-1 and GLUT-2 after a short-term challenge with a high-fat diet in C57BL/6Jmice[J].Diabetes,2002,51:S138-43.
[12] BuShi, Yang, wang, et al. Pancreatic islet long-term high fat feeding of glucose stimulated insulin secretion in rats [J]. The influence of the endocrine metabolism, 2003) : 25-8.
[13] Wang Xiaoling Jiang Lingling XingLiYong, etc. High fat feeding rats with insulin resistance induced dynamic observation [J]. Chinese journal of gerontology, 2008, 4 (28) : 664-664.
[14] child stars, Dong Jiayi WuZhiWei, etc. The long-term high fat feed effects on insulin resistance in rats [J]. Chinese journal of gerontology, 2012, 1 (32) : 70-70.
[15] Li Lanfang Guo Yu Tang Guotao, etc. A high-fat diet led to the mechanism of action of rat hepatic insulin resistance research [J]. Chinese journal of pathology, physiology, 2011, 27 (2) : 310310.
[16] Wu Yanli Zhao Baozhen, XiGuangXia, etc., impaired glucose tolerance Wistar rat model of experimental research. The Chinese medicine and clinical, 2007, 7 (4) : 281-283 [17] Conarello, S. L., et al. Mice lacking dipeptidyl peptidase IV are protected against obesity and insulin resistance. PNAS, 2003,100(11): 6825-6830
[18]ReimerM K,et al.Altered P-cell distribution of pdx-1 and GLUT-2 after a short-term challenge with a high-fat diet In C57BL/6J mice[J].Diabetes,2002,51:S138-43.
[19] Hu Jianping ,Yan Xi, Yang Zhiwei, etc. Diet induced mouse model of metabolic syndrome. Chinese medical journal, 2009, 12 (6) : 34-38.[20]Gianluca SB,Cinzia C,Stefania S,et a1.A Model of Insulin and Nonalcoholic Steatohepatitis in Rats--Role of Peroxisome Prolifertor-Activated Receptor-aand n-3 Polyunsaturated Fatty Acid Treatment on Liver Injury[J].The American Journal of Pathology,2006,169(3):846-860.
[21] Zhou Yunfeng, Li Sha Su Wen, etc. Different fat content is high in fat and purification of formula feed on mice and rats in metabolic syndrome [J]. The influence of basic medicine and clinical, 2012, 32 (3) : 273-277
[22] Xu Lingling, to red, leader, and so on. A high-fat diet induced high blood leptin and insulin in animal models of change [J]. The relationship between basic medicine and clinical, 2001, 21 (5) : 441-441.
[23] Hu Dongsheng. The adult population of type 2 diabetes, epidemiological studies in China. Beijing, Beijing union medical college, 2007.
[24] Zhu Haiqing. Age and family history of Chinese adults the influence of insulin secretion and insulin action [J]. Journal of Beijing, Beijing union medical college, 2012.
[25] Bai Zhenlian Wu Qiuhua. Insulin resistance and related hormone levels test study of middle-aged and old men's health changes along with the age [J]. Journal of taishan medical college, 2007, 28 (4) : 248-248.
[26]BAI Xiu-ping,et al.Sequence of fat partitioning and its relationship with whole body insulin resistance[J].Chinese Medical Journal,2010,123(24):3605-3611.
[27] Tian Aiping, guo shan, ShenZhuFang. High fat feed animal models with insulin resistance [J]. Chinese pharmacological bulletin, 2006, 22 (3) : 267-267.
[28]Kang L,Ayala JE,Lee-Young RS,et al.Diet-induced muscle insulin resistance is associated with extracellular matrix remodeling and interaction with integrin alpha2beta1 in mice[J].Diabetes.2011 Feb;60(2):416-26.
[29] ShenYanJu, Peng Lan Liao Zhiling, etc. STZ replication process of experimental type 2 diabetic rats model and experience summary [J]. J g journal of guangxi medical university, 2011, 28 (6) : 821-821.
A Research about the Influence of Yi Jia Decoction on Serum NO, GSH-PX Levels of Immunological Liver Injury in Mice
12 3
Sui Fangyu Qu Miao Zhang Miao
Heilongjiang University of Chinese Medicine
Chinese Herbal Teaching and Research Section Harbin China
Abstracts: Objective: To explore the functional mechanism of Yi JiaDecoction in treating immunological liver damage in mice through the test on serum NO, GSH-PX levels.
Methods: 120 mice were randomly allocated into six groups which areblank, model, western medicine, high dose of Chinese medicine, medium dose of Chinese medicine and low dose of Chinese medicine groups. Combining BCG and LPS induces liver injury in mice. After two weeks'treatment, mice were testedon serum NO, GSH-PX.
Results: Compared with the model group, serum NO levelof the treatment groups tended to decrease, especially for the medium dose of Chinese medicine group (P<0.01), and GSH-PX level of treatment groups tended to increase, especially for the high dose of Chinese medicine group (P <0.01). Conclusion: Yi Jia Decoction can treat the liver injury in mice and the functional mechanism may be related to the adjustment the serum NO, GSH-PX levels of mice.
Key words: Yi Jia Decoction; Nitric Oxide; Immunological Liver Injury
1 Introduction
Immunological Liver Injury(ILI) is a common and worldwide disease, regardless of race, region and other restrictions. At present, most academics believe that pathological mechanisms of inducing liver injury in mice by BCG and LPS are quite similar to the pathological mechanisms of human hepatitis immune dysfunction. So combination of BCG and LPS is one of the ideal methods to choose and do research on hepatoprotective medicine.[1]Yi Jia Decoction which includes raw oyster shell, raw rhizoma atractylodis macrocephalae, stir-fried barley sprout, schisandra, is a popular formula for treating immunological liver injury. The author chosemice as subjects via BCG and LPS to induce liver injury and exploredthe therapeutic mechanism of ILI.
2 Materials and methods
2.1 Animals and groups
Kunming mice, weighing 18-22g, 60 males and 60 females (All from Heilongjiang University of Chinese Medicine Experimental Animal Center). Experimental environment: Kept the room ventilated, dry, temperature 22-25 °C and humidity50-70%. Made sure that the mice can drink and eat freely. The mice were randomly divided into six groups: blank group, model group, DDB treatment group (western medicine group), high, medium and low dose of Yi Jia Decoction groups and each group included 20 mice.
