Studies On Extraction And Pharmaceuticalaction Of Active Constituent- Paeonol In Cortex Moutan

Diabetes has become the third chronic disease following the tumor and cardiovascular diseases which affected the life quality and health. More than 90% of Diabetes patients belongs to type Ⅱ diabetes. Recently, the combination of muscle insulin injections and oral medications was mainly adopted for clinical treatment.However, it inevitably hurt to the major organs(such as liver, kidney, etc.) too seriously to halt taking medicines due to the widespread adverse reactions and side effects on a body of the synthetic drugs and the requirements to take medicine in liftetime, whereas the method above can reduce the harm from high blood sugar.Natural medicines have received the widespread attention since the small side-effects and no obvious characteristics of adverse reactions in the treatment of diabetes. Paeonol is one kind of traditional Chinese medicines. The present study confirmed that paeonol has obvious therapeutic effect for the model of typeⅡdiabetes,which can not only apparently reduce of blood lipids, blood glucose and hyperinsulinemia in the diabetes model but also make certain curative effect in clinical applications. However, there are still a lot of works need to be solved, such as how to obtain paeonol with the biological activity extracted from plants efficiently and economicly, and how to clearly know the anti-diabetic mechanisms of paeonol. Therefore,this study mainly focus on possible mechanism with paeonol from optimized extraction process of diabetes by using enzyme-linked immunosorbent assay, flow cytometry technique, polymerase reaction, and western blotting methods.This experiment extracted white needle crystal from cortex moutan by combination of the salt water vapor distillation method and Recrystallization method. The extraction rate was 86.40% and the extract was identified as paeonol with purity of 95.21%.In the process of animal experiments, the type Ⅱ diabetes rat model was prepared by adopting streptozotocin in combination with high fat feeding. All of animals were randomly divided into normal group, model group, low-dose group and high-dose group.And the experiments concentrated on the aspect such as the hypoglycemic lipid-lowering,hyperinsulinemia relieving, immunological function adjustment, islet B cell proliferation and differentiation. The results were demonstrated as follows:In diabetic rats group, after different dose lavage treatment for 40 days, blood sugar reduced from the original 16 mmol/ L to 9.8 mmol/L and 7.0 mmol/L, respectively. The level of blood glucose in rats was significantly decreased(p < 0.01) in high dose and low dose groups compared with model group, respectively, serum insulin reduced from 33.75μIU/L to 25.58 and 19.96 μIU/L showing significant reduction(p < 0.01), respectively,which was closed to the value 18.97μIU/L of normal rats. Meanwhile, the weight, blood lipid, glycosylated hemoglobin(Hb Alc) and glucose tolerance of the rats were obviously improved, showing the significent therapeutic effects of paeonol on type Ⅱ diabetes.The results from treatment group and control showed that the immunological function of rats significantly improved and the concentration of characteristic factor of the IL- 2 and TNF alpha of Th1 cells increased in the serum, which stand for the overall activity of Th1 cells. The cellular immunological functionthen was enhenced for the CD8+ cells proliferation differentiation induced by cytokines mentioned above. All these alteration enhanced islet B cell’s ability to resist damage and prevent from B cell’s death.It is demonstrated that the concentration of characteristic factor of Th2 cells and the level of IL-6 gene expression were significantly reduced in the serum based on the test results of immune function changes of rats before and after treatment. It demonstrated that the humoral immunological function induced by Th2 cells is effectively suppressed,and the concentration of Ig and C3 and C4 gradually returned to normal, which reduced the damage to tissues and organs. Paeonol also can increase the serum concentration and gene expression of TGF-beta which is the characteristic factor of Treg cells. By increasing the number and activity of Treg cells and restored the ratio of CD4 + / CD8 +,paeonol improved the body’s innate immunological function.Paeonol can eliminate immune inflammatory reaction caused by diabetes in pancreatic morphology according to the results of HE staining given 40 days’ treatment.At the same time, paeonol can promote the B cell proliferation, differentiation, and participate in repairing and reconstruction of the damaged islet. It was found that the specific mechanism is paeonol can promote PPAR gamma expression significantly in thepancreas tissues, which inhibited the JNK apoptosis signaling pathways and further reduced the injury of the islet B cells. On the other hand, paeonol can enhance expression of GPL-1 in the pancreas tissues, which improves concentration and gene expression of PDX-1, which is a crucial factor to islet B cells which can enhence proliferation and differentiation. And it also participated in the reconstruction and restoration of the damaged islet tissue due to diabetes.In this paper, the anti-diabetic effects of paeonol were deeply studied on a large number of animal experiments adopting advanced molecular biological detection methods. The results provide insight into exploration of the the scientific foundation and theoretical basis of treatment in diabetes.