Mechanism Research Of Rhizoma Anemarrhenae Extracts Improving The Type 3 Diabetes

Rhizoma Anemarrhenae, the dry root and stem of Anemarrhena asphodeloides Bge, which belongs to the family of Liliaceae, the only resource of Traditional Chinese Medicine "zhimu" recorded in the Pharmacopeia of People’s Republic of China (ed.2010), is considered as Zi Yin Run Zao and eliminating heat and purging fire. Modern pharmacological studies have shown the effects of lowing blood sugar, improving cognitive disorders, anti-inflammatory effects et al. Rhizoma Anemarrhenae mainly as saponins, accounting for about 6%; this topic use effective fraction (mainly saponins), from inflammation-cellular stress pathways to explore the mechanisms, the action of lowing blood sugar and improving cognitive disorders. Meanwhile, establishing its quality standards, in order to further its development as a safe, effective and controllable hypoglycemic, improve cognitive disorders drug preparation.Based on current research and preliminary experiments, further research on extracts of Anemarrhenae to determinate methods; qualify standards established and from inflammation-cellular stress pathways to explore the mechanisms to the action of lowing blood sugar and improving cognitive disorders caused by AlCl3.Methods:1 Determination of Anemarrhenae effective fraction contentUse UV, HPLC and HPLC-ELSD, timosaponin BII and Mangiferin as control, to determinate the effective part of the total saponins, total flavonoids, timosaponin BII and Mangiferin content.2 Base on inflammation-cellular stress pathways to explore the mechanisms, the action of lowing blood sugar and improving cognitive disorders caused by AlCl3.64 male SPF SD rat, divided into 8 groups with 8 for eatch group, named as normal control group(NC), diabetes mellitus model group(DM), DM administration group(ZDM), Complex model group(AM), AM administration group(ZAM), Alzheimer’s disease group(AD), AD administration group(ZAD) and Pairacetam group(NK).Intraperitoneal injection of STZ and high fat feed to induce type 2 diabetes models, gavaging AlCl3 induced dementia model and combination of the two modeling methods to induce complex model. Continuously gavage the rats with Anemarrhena effective part for 20 weeks, determinate the serum GLU、TC、TG、INS、Aβ1-42、 TNF-α、p-JNK and other indicators in 8 and 16 week respectively, analyzes the changes and the relationship between type II diabetes and cognitive impairment. At the end of 20 weeks of administration, dissecting and preservatiing the rat brain, the hippocampus and pancreas to made into routine pathological sections and immunohistochemistry for observation.Results:1 The study of Anemarrhena extracts quality standardUse UV Initially established methods for the determination of total saponins and total flavonoids of Anemarrhenae extracts. The result of total saponins and total flavonoids of Anemarrhenae extracts possess of 44.58% and 7.26% respectively. While establishing a HPLC or HPLC-ELSD method for determination of the effective part of timosaponin BⅡI and Mangiferin, timosaponin BII content 15.12%, Mangiferin content 2.82%;2 The research results of Pharmacodynamicsa) 3 weeks later after Injection of STZ,compared with the normal group,the weight of the model group increase significantly, and FPG significantly increased (P<0.05),indicating that type Ⅱ diabetes building success.b) Administration 8 and 16 weeks, FPG, TC and TG level in rats:compared with normal group,blood glucose of DM and AM model group rats increase significantly, have the meaning of meter (P<0.01 or P<0.05);Compared with DM and AM model group,the Anemarrhena extracts has better reduce FPG, TC and TG level (P<0.01 or P<0.05); And the FPG of AD model group has no significant difference, but TC, TG show significant differences (P<0.01 or P<0.05).c) Administration 8 and 16 weeks, compared with normal group, DM,AM and AD model group rats’ blood plasma of Aβ,TNF-α and p-JNK increased significantly, have the meaning of meter (P<0.01 or P<0.05);And with the administration of Anemarrhena extracts, these indicators decreased to normal levels showed significant differences (P <0.01 or P<0.05) to the 3 model groups.d) Administration 20 weeks, compared with normal group, DM,AM and AD model group rats’ hippocampus showed significantly high positive expression of Aβ, TNF-α and p-JNK, have the meaning of meter (P<0.01 or P<0.05); And with the administration of Anemarrhena extracts, the positive expression of these indicators decreased obviously (P<0.01 or P<0.05).e) Pathological section results showed normal pancreatic tissue, hippocampus cells arranged in neat rows, and three models of pancreatic tissue group, hippocampal tissue cells arranged scattered through the administration significantly after repair.Conclusion:a) Initially established the quality standards and determined the material basis of Anemarrhena extracts;b) Anemarrhena extracts showed significant effect of regulating glucose and lipid metabolism, as well as improving cognitive impairment of the three kind of models, intraperitoneal injection of STZ and high fat feed to induce type 2 diabetes models, gavaging AICI3 induced dementia model and combination of the two modeling methods to induce complex model.c) Anemarrhena extracts can significantly reduce the accumulation of Aβ protein, and decreased the expression of TNF-a and p-JNK, indicating Anemarrhena extracts can inhibit inflammation- cell stress pathway activation, and the effect of hypoglycemic, improve recognition cognitive disorders may be through activation of TNF-α/JNK pathway inhibition achieved.