| Objective:Study on the model of diabetic rats after taking Bushen Tongqiao Decoction,to observe the protein expression of TSP-1 and TGF-β1 in hypothalamic neurons and hippocampus. To analyze and prove the influence of the change of TSP-1 and TGF-β1 signaling pathway according to the results. The tentative confirmation is the mechanism of activation of TSP-1 and TGF-β1 signaling pathway in diabetes mellitus complicated with cerebral injury under hyperglycemia state. The result confirmed the effect of Bushen Tongqiao Decoction on cerebral neuronal damage. To explain the mechanism and control principle of diabetic rats after taking Bushen Tongqiao Decoction from molecular biology. It’s the further study for TCM theory and the exact therapeutic effect of traditional Chinese medicine in treatment of diabetes mellitus complicated with cerebral injury. To provide the experimental basis for the treatment of diabetes with nervous system diseases. It’s the basic research for the new drug.Methods:27 clean grade SD male rats were randomly divided 3 groups:Healthy control group, Diabetic model control group, Bushen Tongqiao Decoction treatment group. The rats of Diabetic model control group and Bushen Tongqiao Decoction treatment group were fed with high fat and high glucose diet for one month. After fasting for 12 hours, two groups were treated with intraperitoneal infection 15mg/kg of streptozocin (STZ, to prepare 2% STZ injection by 0.01mmol/L citric acid-sodium citrate buffer solution, PH 4.4). After five days, the two groups were injected by STZ once. The healthy control group was fed with the normal diet and treated with intraperitoneal infection the same dose of O.Olmmol/L citric acid-sodium citrate buffer solution. The Bushen Tongqiao Decoction treatment group was taken by 16g/kg Bushen Tongqiao crude drug decoction once a day from 15 days before modeling to 4 weeks after modeling. All rats were measured glucose in blood and urine once at a fixation time after injection of the third, fifth, tenth, fifteenth, twentieth day. Every group was measured weight once at a fixation time every week. After the detection of blood glucose value for the 28th day, all rats were killed and fixative was injected intravascularly into the brain and tissues were picked up to make into continual coronary paraffin was section. The protein expression and localization of TSP-1 and TGF-β1 in the neurons of hypothalamus and hippocampus area were tested after immunohistochemical staining in immunohistochemical SABC method. Totaling the number of the positive expressions per unit area in the hypothalamus (the nucleus) neurons and hippocampal neurons as their positive cell’s density by objective micrometer. Calculating the fixed position in the groups of the same section neurons in specific site selection of hypothalamus and hippocampus area in unit area in the number of positive cells. Differences between groups were compared by one-way analysis of variance and the differences between groups were compared by t test.Results:①Compared with healthy control group, the reaction and action of the rats of diabetic model control group were slow. The rats were listlessness and the skin lost luster. Their appetite, drinking, diet and urine increased. The weight was lost (P<0.01). The fasting blood glucose levels decreased (P< 0.01).The density of TSP-1 and TGF-β1 positive cells increased in the periventricular hypothalamic nucleus(Pe), paraventricular nucleus(Pa), the anterior area(AH), the lateral area(LH)(P<0.01). The density of TSP-1 and TGF-β1 positive cells increased in hippocampus CA1, CA3(P<0.01).②Compared with diabetic model control group, the reaction and action of the rats of Bushen Tongqiao Decoction treatment group were more flexibily. The rats were good and the skin gloss was better. Their appetite, drinking, diet and urine were normal. The weight was put on (P< 0.01). The density of TSP-1 and TGF-β1 positive cells decreased in the periventricular hypothalamic nucleus(Pe), paraventricular nucleus(Pa), the anterior area(AH), the lateral area(LH)(P< 0.01). The density of TSP-1 and TGF-β1 positive cells decreased in hippocampus CA1, CA3(P<0.01).Conclusion:The expression of Lin28 was closely associated with resistance to paclitaxel. The Lin28/let-7 pathway may be an underlying mechanism of drug resistance of HCC. Our study provides a new research idea for the improvement of chemotherapy in patients with HCC. |
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