| Objective:The purpose of this study is to provide theoretical support for the quality evaluation,development and utilization of the processed P.cyrtonema Hua.,by exploring the hypoglycemic and anti-inflammatory effects of its crude polysaccharides extracted from raw material group and nine-steam-nine-bask processing group.Methods:1.The samples were treated by drying and the traditional processing method of "nine steaming and nine drying" respectively,and the standard curve of glucose was established by ultraviolet spectrophotometry,and the contents of polysaccharides in raw material group and nine-steam-nine-bask processing group were determined by anthrone-sulfuric acid method.2.The model of RAW264.7 cell inflammation was induced by 10 μg/m L of lipopolysaccharide(LPS).The grouping was as follows: blank control group;LPS model group and treated group.Different groups were treated with corresponding methods and cultured in an incubator.The content of NO was detected by Griess method.The m RNA levels of the inflammatory factors TNF-α,IL-1β,IL-6 and MCP1 were detected by qPCR,and the levels of the inflammatory factors TNF-α,IL-1β,IL-6 and MCP1 were detected by ELISA.3.The model of type I diabetes was induced by low-dose intraperitoneal injection of chain urea with streptozotocin(STZ)50 mg/kg for several times,and then the mice whose model had been built successfully were randomly divided into model group(MOG),raw P.cyrtonema Hua.low-dose(3 g/kg)polysaccharides group(RLG),raw P.cyrtonema Hua.high-dose(12 g/kg)polysaccharides group(RHG),nine-steam-nine-bask processing P.cyrtonema Hua.low-dose polysaccharides group(3 g/kg)(NLG),nine-steam-nine-bask processing P.cyrtonema Hua.high-dose polysaccharides group(12 g/kg)(NHG)with the dose calculated according to the amount of raw medicine and positive metformin group(200 mg/kg)(PMG).There wereeight mice for each group,and the administration volume was 0.1 m L/10 g.The mice of MOG were watered as usual,while the other groups’ mice were administered corresponding drugs.Random blood glucose was measured every week.After four weeks,blood samples were taken from the tail tip to measure random blood glucose.Afterwards blood was taken from the orbital vein and then centrifuged.After that serum ALT and AST levels were detected with the kit.The mice were killed by cervical dislocation and their whole livers were taken and weighed.HE,oil red O and glycogen staining sections were prepared from part of the liver.Serum lipid(TG,TC,LDL-c),glucose metabolism-related enzymes(HK,PK),antioxidant enzymes(SOD,MDA)were measured for the rest of liver tissues.Result:1 Anthrone-sulfuric acid method was used in the experiment.The results of glucose standard curve showed a good linear relationship between glucose concentration(0.01～0.1 mg/m L)and absorbance.The polysaccharide content of raw materials P.cyrtonema Hua.was 7.34% and nine-steam-nine-bask processing group3.48%.Compared with raw products,the content of polysaccharides in nine-steam-nine-bask processing products decreased significantly.The results showed that the polysaccharides content of P.cyrtonema Hua.could decrease in the steaming process with a reduction rate of 52.6%.2 The effect of polysaccharide concentration on cell viability was determined by cck-8.The results showed that in the range of 125-2000 μg/m L,125-500 μg/m L of polysaccharide of raw materials and nine-steam-nine-bask processing products had no side effects on cell survival and the cell morphology remains good.Therefore,125-500μg/m L was selected as the administration concentration.Within the range of 0.1 μg/m L ～50μg/m L LPS administrated,the results of Griess showed that compared with the normal group,the morphological changes were obvious after the cells were treated with 10 μg/m L LPS,and at the same time,the content of NO in the supernatant was the highest,so 10 μg/m L was used to establish inflammatorymodel.The levels of NO in LPS-induced RAW264.7 inflammatory cells were then detected for groups treated with different concentrations of raw materials and nine-steam-nine-bask processing P.cyrtonema.The results showed that the content of NO in the model group was significantly higher than that in the normal group.Compared with MOG,the NO contents in the group of different concentrations of polysaccharides raw materials and nine-steam-nine-bask processing P.cyrtonema were significantly reduced and showed a concentration dependence.There was NO significant difference regarding NO between nine-steam-nine-bask processing groups and raw material groups treated with same doses of polysaccharides.ELISA results showed that,compared with the normal group,the contents of TNF-α,IL-1β,IL-6 and MCP1 in the supernatant of the model group were significantly increased.Compared with MOG,the levels of TNF-α,IL-1β,IL-6 and MCP1 in groups with different concentrations of polysaccharides in raw materials and nine-steam-nine-bask processing P.cyrtonema were significantly reduced in a concentration-dependent way.There was no significant difference in trems of TNF-α,IL-1β,IL-6 and MCP1 between nine-steam-nine-bask processing groups and raw material groups treated with same doses of polysaccharide.qPCR results showed that compared with the normal group,the levels of TNF-α,IL-1β,IL-6 and MCP1 m RNA in the supernatant of MOG were significantly increased.Compared with MOG,the levels of TNF-α,IL-1β,IL-6 and MCP1 m RNA in the treated groups with different concentrations of polysaccharides were significantly reduced in a concentration-dependent way.There was no significant difference concerning m RNA levels of TNF-α,IL-1β,IL-6 and MCP1 between nine-steam-nine-bask processing groups and raw material groups treated with same doses of polysaccharides.3 Compared with the normal group,the STZ-induced diabetic mice had obvious characteristics of "three more and one less",and random blood glucose concentration was ≥ 16.7mmol/L and it indicated that the diabetes model was successfully established.Compared with MOG,the high-dose group of raw materials and nine-steam-nine-baskprocessing group had certain hypoglycemic effect.Compared with the raw material polysaccharide groups,nine-steam-nine-bask processing polysaccharide groups did not show differences with regard to inhibitory effect on blood glucose.Compared with MOG,AST and ALT levels in the high-dose serum of raw material groups and nine-steam-nine-bask processing groups decreased significantly.Compared with the raw material polysaccharide groups,there was no difference in the inhibition of ALT,AST and liver index.Compared with MOG,TG,TC and LDL-C were significantly reduced in the liver of the high-dose groups of raw and nine-steam-nine-bask processing P.cyrtonema.Compared with raw material polysaccharide groups,nine-steam-nine-bask processing groups showed no difference regarding TG and LDL-C,and significant differences concerning TC.Compared with MOG,the content of SOD and MDA in the liver of high dose group of raw material and nine-steam-nine-bask processing group increased significantly,and the content of MDA decreased significantly.Compared with raw material polysaccharide groups,nine-steam-nine-bask processing groups showed no difference with regard to SOD and MDA.Compared with MOG,the activity of HK and PK enzymes in the liver of high dose group of raw material and nine-steam-nine-bask processing group increased significantly.Compared with raw material polysaccharide groups,nine-steam-nine-bask processing groups showed no difference concerning HK and PK.Compared with the normal group,the liver steatosis was severe in MOG,and the improvement effect was obvious in the high dose group of raw material and nine-steam-nine-bask processing group.Compared with the liver glycogen content in the model group,the glycogen content in the liver of high dose group of raw material and nine-steam-nine-bask processing group increased significantly.Compared with raw material polysaccharide groups,nine-steam-nine-bask processing groups showed no difference with regard to storage capacity of liver glycogen.Conclusion:1 During processing,the content of polysaccharides decreased due to the decomposition of water-soluble polysaccharides-mucinous.2 Polysaccharides from raw materials and nine-steam-nine-bask processing P.cyrtonema demonstrated no toxicity and side effects on LPS-induced RAW264.7 cells,and showed obvious inhibitory effects on the inflammatory cytokines NO,TNF-α,IL-1β,IL-6 and MCP1 in a dose-dependent manner.Thus,it is assumable that polysaccharides from raw materials and nine-steam-nine-bask processing P.cyrtonema play an anti-inflammatory role by inhibiting the expression of related inflammatory factors.3 Polysaccharides of raw materials and nine-steam-nine-bask processing P.cyrtonema have certain protective effects on type Ⅰ diabetic liver.It may be related to the regulation of glucose and lipid metabolism(TC,TG,LDL-C,PK,HK),antioxidant(SOD,MDA),liver function(ALT,AST),etc.Moreover,the protective effect of polysaccharides from nine-steam-nine-bask processing P.cyrtonema on liver was equivalent to that of polysaccharides from raw materials.The results indicated that polysaccharides of raw materials and nine-steam-nine-bask processing P.cyrtonema could play a good hypoglycemic role by protecting the liver.4 Rhizoma polygonati is a common medicinal material for both food and medicine.The research direction should pay more attention to whether it will bring adverse effects to the body after being taken.After steaming,P.cyrtonema reduces irritation and is more conducive to taking,so it can be shown that in the practical application of anti-inflammatory and hypoglycemic,nine-steam-nine-bask processing P.cyrtonema polysaccharide has a better performance. |
PDF Full Text Request