| Background: 2 diabetes is caused by a variety of causes metabolic disorders, characterized by chronic hyperglycemia. Although the current study did not fully clarify the pathogenesis of type 2 diabetes, but it is recognized as an important part of both insulin resistance and β cell secretory dysfunction, including type 2 diabetes throughout the whole process is the development of insulin resistance. According to the research results in recent years, play an important role in insulin resistance in type 2 diabetes is inflammation and lipid metabolism. Glucose or lipid simply no longer the ultimate goal of diabetes treatment for sugar, fat, represented by a cascade of problems is the top priority for the comprehensive regulation. The ultimate goal of medicine, mostly single adjustment mechanism, are more prone to the phenomenon of increased glucose and body weight(such as insulin sensitizers) simultaneously, in addition to lipid lowering, but also lead to liver damage and other defects. In this paper, experimental study, literature review, theoretical discussion and other methods of traditional Chinese medicine bitter acid through transfer method, even plum decoction prevention of type 2 diabetes and lipid metabolism were studied systematically.Relationship between obesity and inflammation, lipid metabolism and insulin resistance in type 2 diabetes: obesity and type 2 diabetes and insulin resistance are closely linked, especially central obesity. Obesity is common in patients with type 2 diabetes problem, according to the results of recent epidemiological survey, 85%-90% of patients with type 2 diabetes are overweight or obese. Obesity is closely associated with chronic inflammation, obesity, adipose tissue is the main cause of chronic inflammation of the initiation site, especially visceral adipose tissue in the inflammatory response produce and played a very important role in regulation.Of insulin signal transduction generated resistance, a variety of inflammatory cytokines are secreted by fat cells obstruct, and thus lead to the occurrence of type 2 diabetes. Thus, obesity and inflammation, lipid metabolism, insulin resistance and type 2 diabetes are closely related. So in order to be more effective in the prevention and treatment of obesity, type 2 diabetes, obesity need to be clear, the molecular mechanisms of inflammation, lipid metabolism and insulin resistance.Although a large number of in vivo experiments on by inflammation, lipid metabolism and insulin resistance, inflammation, we have preliminarily confirmed the important role of lipid metabolism in the pathogenesis of obesity, insulin resistance and type 2 diabetes have played in previous work, but not yet inflammation, lipid metabolism and insulin resistance linked.SREBPs is a class located in the endoplasmic reticulum membrane connexin, is a nuclear transcription factor, SREBPs synthesized in the endoplasmic reticulum and SREBP cleavage activating protein(SCAP) binding to the Golgi apparatus by which, in the Golgi apparatus were site 1 and site 2 protease protease sequence processing, the final release of the amino-terminal fragment into the nucleus and its target gene promoters of sterol regulatory element binding,regulate the expression of related genes.SREBP-1c play an important role in the development of lipid metabolism i.e. an accumulation of lipid in the process. SREBP-1c, also known as adipocyte differentiation factor 1, mainly in rodents and fat cells expressing human liver and involved in the regulation of fatty acid, triglyceride synthesis and expression of glucose metabolism-related gene. SREBP-1c in adipose tissue-specific expression, the liver to insulin play a role in protein synthesis is limited, so that the decline in insulin action, causing insulin resistance and hyperinsulinemia; In addition, lipid accumulation process is mature fat cell volume increasing process, with lipid synthesis(triglycerides, fatty acid synthesis) gene expression associated correlation. Regulated by SREBP-1c FAS play an important role in fatty acid synthesis. Free fatty acids(free fatty acids, FFA) as an intermediate product of fat metabolism, it can reduce the increase in insulin sensitivity, which can cause or worsen insulin resistance. Therefore, by preventing and reversing SREBP- 1c expression may be an effective way of prevention and treatment of type 2 diabetes and its complications.PART1.Side impact bitter acid pass notes on glucose and lipid metabolism in experimental diabetic ratsPurpose:Diabetes is characterized by high blood sugar, and also associated with mutual influence lipid metabolism, blood sugar and blood lipids. Therefore, the treatment of active and effective in regulating blood sugar in diabetic patients, it should also pay attention to regulate lipid metabolism. A research-based experimental basis, through the general state of experimental rats, blood glucose, lipids, insulin levels and other tests to investigate the effect of bitter acid side pass notes on glucose and lipid metabolism in diabetic rats.Method: 1. experimental animal model The 42 and 10 ZL rats ZDF rats were divided into cages, 2 per cage, free feeding, drinking water. Each group was given normal diet adaptive feeding one week later, ZL group continue to give normal diet, ZDF group was given high-fat diet. To 12 weeks, a total of 30 diabetic ZDF rats.2. The judge diabetic rats modeling success criteria Regular line tolerance test in rats, fasted before the test water 15 h, after cutting the tail blood glucose meter trace fasting glucose, 30% glucose solution according to 2g / kg orally. After glucose load were at 0.5h, 1h, 1.5h and 2h tails blood, with blood glucose meter measurements. When blood glucose peak is higher than 16.7mmol / L, and glucose loading 2h higher than 11.1mmol / L, can be diagnosed as diabetes. When any one of these, if any, for impaired glucose tolerance. Take a diagnosis of diabetes or impaired glucose tolerance in rats as experimental subjects.3. Animal grouping and method of administrationThe successful model 30 ZDF rats were randomly divided into model group(n = 10), pass notes bitter acid group(n = 10), pioglitazone group(n = 10) 3 group, 10 rats were used as blank control ZL groups. The bitter acid side pass notes and pioglitazone are configured into a suspension, after the model is successful, bitter acid side pass notes when used in accordance with 3.29 g / kg / d of body weight(preliminary work has identified the optimal dose) suspension gavage, pioglitazone group, according to 1.07 mg / kg / d of body weight orally, blank control group and model group herein are an equal volume of distilled water, the rats were intragastrically 12 weeks, 1 day. During drug treatment control group of rats fed a normal diet, the other groups were fed with high fat diet.Result: 1.general During feeding, ZL rats than in active resistance to crawl obvious, normal eating drinking, glossy coat. ZDF rats compared with ZL rats obesity, weight rapidly, activity slow reactions are blunt, dry and dull coat, each group had no deaths.2 weight change Before and after treatment changes in body weight of rats in each group as shown in Table 1. After drug treatment, the model group, pass notes bitter acid group, pioglitazone group, body weight was significantly higher than the control group(P <0.01). After four weeks of treatment, body weight each treatment group compared with the model group, although the downward trend, but not statistically significant(P> 0.05). After 12 weeks of treatment, treated rats decreased body weight compared with the model group was significantly(P <0.01), bitter acid pass notes rats body weight decreased significantly compared with the model group(P <0.05), body weight of rats pioglitazone group has decreased compared to the untreated group, but no significant difference(P> 0.05).3 blood glucose 3.1 Changes in rat blood glucose OGTT After 12 weeks of treatment, compared with the normal control group, model group, pioglitazone group, pass notes bitter acid group was significantly higher fasting glucose, a significant difference(P <0.01). Treatment group were significantly lower blood sugar than the model group(P <0.01), pass notes bitter acid blood glucose compared with pioglitazone there was a significant difference(P <0.01).3.2 serum insulin(INS)(That is, after 12 weeks of treatment) in rats at 34 weeks of age, compared with the normal control group, model group, serum insulin levels were significantly increased(all P <0.01). Compared with the model group, pioglitazone group, bitter acid pass notes serum insulin has a different degree of reduction, there were significant differences(P <0.01), but there was no significant difference(P compared to the bitter acid pass notes group and pioglitazone group > 0.05).3.3 glycated hemoglobin(Hb A1C) In the 34-week-old rats(ie, 12 weeks after treatment), compared with the normal control group, model group were significantly higher glycated hemoglobin, there are significant differences(P <0.01). Compared with the model group, pass notes bitter acid group Hb A1 C decreased significantly(P <0.01), no significant difference(P pioglitazone group compared with the model group> 0.05).4.lipids After 12 weeks of treatment test results showed that, compared with the normal group, model group, serum TG, TC, FFA levels were significantly higher(P <0.01). Compared with the model group, pioglitazone group, pass notes bitter acid group TG, TC, FFA was significantly lower(P <0.05, P <0.01). Pass notes bitter acid group compared with pioglitazone group, lower TG, TC, there was a significant difference(P <0.05, P <0.01) FFA levels. Bitter acid pass notes show party can reduce diabetic rats with experimental serum TC, TG, FFA levels, there regulate lipid metabolism.Conclusion:1.can only pass notes bitter acid significantly reduced blood sugar ZDF rats, glycated hemoglobin levels and serum insulin levels, suggesting that bitter acid pass notes can only increase insulin sensitivity, improve insulin resistance.2. can only pass notes bitter acid significantly reduced TG, TC and FFA levels ZDF rats, suggesting that bitter acid pass notes party has a better regulate the role of lipid metabolism.Part2.Side impact bitter acid pass notes to adipose tissue m RNA expression in experimental diabetic rats of FasPurpose: To observe the bitter side of the case pass notes acid-abdominal fat tissue FAS m RNA expression in experimental diabetic rats, and to explore the bitter tone Treating acid through a new target type 2 diabetes.Method: 1 Grouping and administration(with the experimental one)2 Specimen Collection Rats at 12 weeks after treatment ended gavage, fasted water 12 h, the rats were anesthetized with ether, open the abdominal cavity, rapid extraction of abdominal fat tissue, rapidly into ammonia tank after packing to move to-80 ℃ frozen in the fridge for later use.3.Expression 3 RT-PCR method to detect the Fas m RNA in adipose tissue of rats 3.1 Extraction of rat abdominal adipose tissue total RNA 3.2 RNA purity determinationAfter the adipose tissue total RNA was extracted using Trizol method, using UV spectrophotometer 260 nm, 280 nm UV absorbance, repeated three times. Each set of results as A260 / A280 between 1.8-2.0, indicating a higher purity of the extracted RNA.3.3 RNA reverse transcription reaction(1) take 0.2ml EP tube, the following reagents were added to complete the configuration of the RT reaction solution(2) The above reagent mix stand for 10 minutes, the PCR instrument reaction(45C water bath 40 min, 95 ° water bath for 5min, 5C water bath 5min) after inactivation of reverse transcriptase, stored at-20 c refrigerator spare.3.4 RT-PCR reaction 3.4.1 primer sequences(see Table 6) 3.4.2 The reaction conditions(1) GAPDH reaction conditions(2) Fas reaction conditions 3.5 PCR detection and analysis productsResult: The expression of Fas m RNA in adipose tissue of rats By RT-PCR method to detect the expression levels of abdominal fat tissue in rats of Fas m RNA. The results showed that 12 weeks of treatment, the expression of the model group, pioglitazone group, bitter acid pass notes abdominal fat tissue in rats Fasm RNA with the control group was significantly higher than(P <0.01). Bitter acid pass notes group and pioglitazone group compared with model group were significantly lower(P <0.01, P <0.05), described the bitter acid pass notes parties and pioglitazone reduce the role of Fas gene-abdominal adipose tissue of diabetic rats. Pass notes bitter acid group than pioglitazone group(P <0.05). The results shown in Figure 1.Conclusion: The expression of Fas m RNA ZDF rat adipose tissue was significantly upregulated by the expression of Fas m RNA in rat adipose tissue through Callees bitter acid significantly reduced after treatment, suggesting that side can pass notes bitter acids inhibit the expression of Fas, Fas consider suppression expression may be one of the mechanisms through bitter sour tone square regulate lipid metabolism.PART3.Side impact bitter acid pass notes to SCAP SREBP-1c m RNA and protein expression in adipose tissue of experimental diabetic rats.Purpose:Observe the bitter side of the case pass notes sour expression of experimental abdominal adipose tissue in diabetic rats SCAP, SREBP-1cm RNA and protein explore bitter acid pass notes Treatment of type 2 diabetes a new targetMethod: 1. Grouping and administration(with the experimental one)2. Specimen collection Rats at 12 weeks after treatment ended gavage, fasted water 12 h, the rats were anesthetized with ether, open the abdominal cavity, rapid extraction of abdominal fat tissue, rapidly into ammonia tank after packing to move to-80 ℃ frozen in the fridge for later use.3. RT-PCR was used to detect the expression of adipose tissue of rats SCAPm RNA, SREBP-1c m RNA in 3.1 rat peritoneal fat tissue extraction of total RNA 3.2 RNA purity determination 3.3 RNA reverse transcription reaction 3.4 RT-PCR reaction 3.4.1 The reaction conditions 3.5 PCR detection and analysis products4 Western-blot expression was detected in adipose tissue of rats SCAPm RNA, SREBP-1c protein 4.1 Protein samples were collected 4.2 Electrophoresis 4.2.1 SDS-PAGE gel formulation 4.2.2 transmembrane 4.2.3 Closed4.2.4 HybridResult: 1. The expression in adipose tissue of rats in each group SCAP, SREBP-1c proteinBy Western blot analysis to detect the expression levels of adipose tissue in rats SCAP, SREBP-1c protein. The results showed that 12 weeks of treatment, compared with the control group, model group, pioglitazone group, the SCAP adipose tissue through bitter sour tone group, the expression levels of SREBP-1c protein was significantly increased(P <0.01). Significantly reduce the pain of acid through transfer group and pioglitazone group compared with model group(P <0.01, P <0.05), described the bitter acid pass notes parties and pioglitazone reduced adipose tissue in diabetic rats SCAP, the expression levels of SREBP-1c protein. Pass notes bitter acid group than pioglitazone group(P <0.05). The results shown in Figure 2.2. expression in adipose tissue of rats in each group SCAP, SREBP-1c m RNA in By RT-PCR method to detect the expression levels of the SCAP abdominal fat tissue of rats, SREBP-1c m RNA’s. The results showed that 12 weeks of treatment, the SCAP model group, pioglitazone group, bitter acid pass notes rats abdominal fat tissue, SREBP-1c m RNA expression was significantly higher than with the normal control group(P <0.01). Significantly reduce the pain of acid through transfer group and pioglitazone group compared with model group(P<0.01, P <0.05), described the bitter acid pass notes parties and pioglitazone reduce the role of abdominal fat tissue of diabetic rats SCAP, SREBP-1c gene expression. Pass notes bitter acid group than pioglitazone group(P <0.05). The results shown in Figure 3.Conclusion: ZDF rat adipose tissue SCAP, the expression of SREBP-1c protein was significantly increased by the adipose tissue bitter acid side pass notes after treatment SCAP, the expression of SREBP-1c protein was significantly decreased, suggesting that the bitter side to suppress acid pass notes SCAP and SREBP-1c combined suggesting that inhibition of the expression of SCAP / SREBP pathway gene and protein pathway may be one of the traditional Chinese medicine. |
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