| Literature researchDiabetes mellitus is a chronic comprehensive disease which is formed under the common action of gene, eating habits, lifestyle, environment and other factors. Diabeticmacroangiopathy is a common chronic complications of diabetes mellitus, which is the main cause of death and disability in diabetes. AS is the main pathological change of diabetic macroangiopathy. Therefore, preventing and delaying the occurrence and development of diabetic macroangiopathyhave a very important social and economic value.It is considered that the NF-κB pathway is closely related to inflammatory, which is a key pathogenesis of diabetic macroangiopathy.As western medicine is of many limitations, it is necessary to figure out more effective treatments and medicines. Traditional Chinese Medicine has a long history for treating diabetes. Syndromes of diabetes and its vascular complications had already been recorded in "Huang Di Nei Jing". "Dryness heat due to deficiency of Yin" is the traditional understanding of diabetes, but when it comes to modern people’s lifestyle, we consider "Yin deficiency and Yang depression" is another pathogenesis of diabetic macroangiopathy. Xiong Manqi, professor of Guangzhou University of Chinese Medicine, with her research team,had developed a more effective decoction called Modified Taohe Chengqi for preventing complications of T2DM, which was based on traditional Taohi Cheng Qi Decoction. Moreover, part of the mechanism of Modified Taohe Chengqi Decoction had been confirmed by many studies. Experimental study: ObjectiveBase on theory of purgating Yang, my aim is to figure out the TCM mechanism of Modified Taohe Chengqi Decoction for preventing diabetic macroangiopathy. Secondly, based on inflammation and NF-κB signaling pathway, by usinganimal experiment and modern biological technology, we tried to explore the molecular mechanism of Modified Taohe Cheng Qi Decoction in treating diabetic macroangiopathy, which was to figure out the best compatibility of Modified Taohe Cheng Qi Decoction and expand the scope of application.Methods120 male SD rats were firstly fed for 1 weeks, thenlO of them were randomly divided into the normal group.which were fed with normal diet. The rest were chosen as prepared model group, which were fed with high-fat diet. After 6 weeks,21 rats were randomly chosen from the model group, and randomly divided into 3 groups. Each group was injected STZ by 35mg/kg,40 and 45 mg/kg dose.72 h after measuring tail fasting blood glucose, those FPG above 11.1 mmol/L and accompanied with DM symptoms were defined as successful T2DM model. Then through summarizing the condition of successful T2DM rats, we found out 40 mg/kg dose of STZ was the best dose. Thus we injected the rest with 40 mg/kg dose of STZ while the normal group rats were injected with equal amount of citric acid.74 T2DM model rats were finally successfully made. Then the 74 T2DM model rats were randomly divided into T2DM model group, metformin group, Modified Taohe Chengqi Decoction group, chemical party 1 group, chemical party 2 group. The Metformin group were treated by 0.156g/kg dose of metformin. Other group except T2DM model group were treated with 5.4g/kg dose of corresponding Chinese medicine. The normal group and T2DM model group were treated with distilled water. The total intervening time were 8 weeks. We checked the general situation such as weight, food intake, water intake and FPG every 4 weeks. At the end of the experiment, Fins, HOMA IR, HBA1C, TC, TG, HDL-C, LDL-C, IL-6, TNF-a, were measured by ELISA method. Aorta pathology morphology were observed by light microscopy. NF-k B, MCP-1 and VCAM-1 were measured by immuno histo chemical method.Western blotting was used to detect NF-κB, VCAM-1 protein expression. RTQ-PCR was to detect the mRNA expression of NF-κB, MCP-1 and VCAM-1.results1. Compared to the model group, after4 weeks of treatment, the interve ntional group were improved in body weight, water intake and food intake (P<0.01), andthere were no significant difference between the groups (P > 0.05); After 8 weeks of treatment, as to body weight and water intake, there were no significant difference between chemical party 2 group and metformin group (P>0.05) while there were significant difference between chemical party 2 group and original formula group, chemical party 1 grou p, positiv (P< 0.01), as well as chemical party 1 group (P< 0.01);Howe ver, there were also no significant difference among original formula gro up, chemical party 1 group and chemical party 2 group (P>0.05)2. As to glucose and lipid metabolism, after 4 weeks of treatment, compared to the model group, the interventional group were reduced in FBG (P< 0.01), but there were no significant difference between chemical party 1 group and metformin group (P>0.05).After 8 weeks of treatment, chemical party 2 group decreased significantly in FBG compared to original formula group and chemical party 1 group (P< 0.01) but where were no significant difference when compared to metformin group (P> 0.05). As to HBAlc, compared to model group, original fomula group, chemical party 1 group, chemical party 2 group and metformin group had decreased significantly (P< 0.01), but there were no significant difference between chemical party 2 group and metformin group.Both chemical party 2 group and metformin group had decreased significantly when compared to original fomula group and chemical party 1 group (P< 0.01). As to Homa-IR, compared to model group, original fomula group, chemical party 1 group, chemical party 2 group and metformin group had decreased significantly (P< 0.01) and there were no significant difference between metformin group and chemical party 1 group. But metformin group had . decreased significantly when compared to original fomula group and chemical party 2 group.As to blood lipid, compared to the model group, the interventional group were significantly decreased in TC, TG, LDL-C (P< 0.01) and increased significantly in HDL-C(P< 0.01); Compared to the normal control group, the interventional group had significantly increased in TC, TG(P< 0.01). And there were no significant difference among chemical party 2 group, chemical party 1 group and the original fomulagroup(P> 0.05). Besides, there were no significant difference between chemical party 2 group and metformin group (P> 0.05).3. As to the level of blood inflammatory factors, after 8 weeks of treatment, compared to model group, the interventional group had improved significantly in IL-6 and TNF-α(P<0.01), but there were no significant difference in IL-6, TNF-α among metformin group, chemical party 1 group and chemical party2 group (P>0.05) while they were decreased significantly when compared to the original formula group (P<0.01).4. As to aorta pathology morphology, there were no pathological changes in normal group while the model group had showed significantly proliferation in smooth muscle cell and fibrous cap in intimal surface uplift. The fourinterventional groups had showed effect on delaying pathology changes in aortaand it seemed that there were no significance between the chemical party 2 group and metformin group.5. As to VCAM-1, MCP-1, NF-κB protein expression:immunohis to chemical results showed:compared to the normal group, VCAM-1, MCP-1, NF-κB increased significantly in model group, which were presented brown granule sediments, deeper color, and higher level of IOD value (P<0.01); VCAM-1 mainly expressed in the endothelial layerand MCP-1 mainly expressed in the smooth muscle layer, while NF-κ Bexpressed both in ndothelial layer and smooth muscle layer; Compared to the model group, VCAM-1, MCP-1, NF-κB in interventional reduced significantly with a lighter color and lower IOD values (P< 0.01).As to protein expression between interventional groups:①there were significant difference between chemical party 1 groupand original formula group in IOD value (P<0.01);Compared to chemical paryt 2 group, metformin group showed no significant difference in IOD value (P> 0.05), but the two group reduced significantly when compared to original formula group and chemical party 1 group (P<0.01). ② As to MCP-1 protein expression between interventional groups, chemical party 1 group reduced significantly in IOD values when compared to original formula group (P< 0.05), Chemical party 2 group showed no significant difference when compared to metformin group (P> 0.05), but both of them reduced significantly when conmpared to chemical party 1 group and original formula group (P<0.01). ③As to NF-κB protein expression, there were no significant difference between original formula group and chemical party 1 group (P> 0.05); Chemical party 2 group and metformin group reduced significantly when compared to original formula group and chemical party 1 group (P< 0.05), but the two showed no significant difference(P>0.05). As to western blotting protein measurement, compared to normal group, model group, NF-κB, VCAM-1 in other group increased significantly(P< 0.01); Compared to model group, NF-KB, VCAM-1 protein expression in each interventional group were significantly decreased (P< 0.05); Chemical party 2 group and metformin group reduced significantly when compared to original formula group and chemical party 1 group (P< 0.05),but the two showed no significant difference(P>0.05). Besides, there were no significant difference between original formula group and chemical party 1 group (P< 0.05).6.The result of RTQ-PCR showed:Compared to normal group, VCAM-1, MCP-1, NF-KB mRNA expression in other group were significantly increased (P<0.01); Compared to model group, the interventional groups reduced significantly (P <0.01). Chemical party 2 group reduced significantly in NF-K BmRNA when compared to original formula groupand chemical party 1 group(P<0.01) and decreased (P<0.05) when compared to the metformin group. And metformin group reduced significantly when compared to original formula group and chemical party 1 group(P<0.01);Chemical party 2 group reduced significantly in MCP-1 mRNA expression when compared to original formula group, chemical party 1 groupand metformin group(P<0.01). There were no significant difference among the above three group (P>0.05); As to VCAM-1 expression, the chemical party 1 group, metformin group, chemical party 2 group, reduced significantly when compared to original formula group (P< 0.01). Chemical party 2 group reduced significantly compared to chemical party 1 group and metformin group (P< 0.01). There were no significant difference between chemical party 1 group and metformin group(P>0.05).Conelusion1. Based on purgating Yang theoryand "Yin deficiency and Yang depression" theory of the pathogenesis of diabetic macroangiopathy, Modified Taohe Chengqi Decoction caneffectively prevent diabetic vascular complications by their purgating Yang effect.2. Modified Taohe Chengqi Decoction can inhibit the inflammation of type 2 diabetes mellitus and prevent of macrovascular complications. The mechanism might be through inhibiting NF-κB expression and reducing IL-6 and TNF-α in serum, thus inhibiting the expression of MCP-1 and VCAM-1 mRNA and protein. In addition, this study also figuring that, Modified Jiawei Taohe Cheng Qi Decoction had better effect on inhibiting inflammationmainly through adding the dose of Ramulus Cinnamomi, and adding Herba asari. |
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