| Objective1.HPLC fingerprint of sijunzi decoction and a method for simultaneous determination of 6 components was established.2.Evaluation of Sijunzi decoction gradient ethanol extract adjustment by high sugar and high fat diet combined with intraperitoneal injection of low dose(40 mg·kg-1)streptozotocin?STZ?induced?type diabetes mellitus?T2DM?mice glucolipid metabolic disorders.3.Evaluation of sijunzi decoction and different extracts on?type diabetes mellitus?T2DM?intervention of renal injury in rats.Methods1.Research on quality controlThe separation was performed on a chromatographic Diamonsil C188 column?250 mm×4.6 mm,5?m?,with acetonitrile-0.05%phosphoric acid as the mobile phase for gradient elution.Column temperature was 25?;Volume flow rate was 0.5 mL/min.Injection was 10?L and the detection wavelength was 220 nm.2.Resisting?diabetes mice ethanol parts selectionThe T2DM mice model was induced by high sugar and high fat diet for 4 weeks combined with low dose of STZ intraperitoneal injection.ICR mice were randomly divided into blank group?KBZ?,model group?MXZ?,metformin hydrochloride group?ERJSGZ?,Xiaokeping tablet group?XKPPZ?,Sijunzi decoction 0%ethanol extraction group?SJZT0%YCZ?,Sijunzi decoction 30%ethanol extraction group?SJZT30%YCZ?,Sijunzi decoction 60%ethanol extraction group?SJZT60%YCZ?and Sijunzi decoction90%ethanol extraction group?SJZT90%YCZ?.Measurements including body weight in mice,eating,drinking,each viscera index?heart,liver,spleen,lung,kidney,pancreas?,glucose tolerance test,fasting blood glucose?FBG?,fasting insulin?FINS?,insulin resistance index?HOMA IR?,triglycerides?TG?,total cholesterol?T-CHO?content,low density lipoprotein cholesterol?LDL-C?content,high density lipoprotein cholesterol?HDL-C?content and?LG?of hepatic glycogen content.3.Research on intervention?diabetes rat kidney damage assessmentRat were treated with ip injection of streptozocin?STZ?after having high-glucose and high-fat diet for 4 weeks to induce type diabets,rat were randomly divided into blank group?KBZ?,model group?MXZ?,positive drug group?YXYZ?,the Sijunzi Decoction water extract group?SJZTSZ?,Water extract alcohol precipitation group?SJZTSCWZ?and water extract alcohol precipitation supernatant group?SJZTCSYZ?.The indicators included general behavior,body weight,24h urine volume,renal coefficient,blood glucose?FBG?,urine protein?PRO?,blood creatinine?SCr?,urea nitrogen?BUN?and HE stained renal histopathological sections.Results1.Research on quality controlThe fingerprint of Sijunzi Decoction extractum was established.There were 24common peaks marked in total,identified uridine,liguiritin,lobetyolin,liquritigenin,glycyrrhizic acid and Atractylenolide III 6 kinds of ingredients,peak 1 from Codonopsis pilosula,Atractylodes macrocephala and Poria cocos,peak 2 from Codonopsis pilosula and Poria cocos,peaks3,4 and 6,10 from Codonopsis pilosula,peak 22 from Atractylodes macrocephala and peak 5,79,1121,23,24 from Glycyrrhiza uralensis.The method of simultaneous determination content was established of the uridine,liguiritin,lobetyolin,liquritigenin,glycyrrhizic acid and Atractylenolide III 6 kinds of components,it has a good linear relationship with the peak area in the range of 0.13381.338?g?r=0.9996?,0.0870.87?g?r=0.9999?,0.08840.884?g?r=0.9998?,0.08060.806?g?r=0.9998?,0.17761.776?g?r=0.9999?,and 0.08120.812?g?r=0.9999?,respectively;RSD values of precision,stability and repeatability were all less than 3.0%.The average recovery rates of the investigated compounds were 100.39%,99.40%,97.62%,97.00%,99.64%and99.00%,respectively,and the RSD values were 1.80%,1.15%,2.15%,1.29%,1.17%,2.01%,respectively.2.Resisting?diabetes mice ethanol parts selectionCompared with KBZ,the body weight in MXZ was no significant difference,the amount of food,drinking water,AUC,FBG,FINS,HOMA-IR,TG and T-CHO increased significantly?P<0.01?,HDL-C decreased significantly?P<0.05?,LDL-C increased significantly?P<0.05?,and LG decreased significantly?P<0.01?.The viscera coefficient increased significantly only in liver?P<0.01?,but there were no other statistical differences.Compared with MXZ,there were no significant difference in body weight and viscera coefficient in each treatment group,the amount of food,AUC,FBG and HOMA-IR were decreased?P<0.01?,LG increased significantly?P<0.01?.The amount of drinking water in ERJSGZ were significantly decreased?P<0.01?,while the other groups were significantly decreased?P<0.05?,the FINS content of SJZT0%YCZ was significantly decreased?P<0.05?,while the other groups were significantly decreased?P<0.01?.TG and T-CHO contents in ERJSGZ showed no significant difference,but decreased significantly in other groups?P<0.05?.LDL-C content of XKPPZ,SJZT60%YCZ and SJZT90%YCZ decreased significantly?P<0.05?,and there were no statistical difference between the other groups.There were no significant difference in HDL-C content between ERJSGZ and SJZT0%YCZ,but the other groups were significantly increased?P<0.05?.3.Research on intervention?diabetes rat kidney damage assessmentCompared with KBZ,the general behavior of the rats in MXZ was obviously abnormal,the weight was significantly decreased?p<0.01?,urine volume and renal coefficient were significantly increased?p<0.01?,FBG,SCr,BUN were significantly increased?p<0.01?,PRO significantly increased?p<0.05?,renal pathological damage was obvious.Compared with MXZ,the general behavior of the drug groups were significantly improved;urine volume and renal coefficient of YXYZ were significantly decreased?p<0.01?,the other drug groups were decreased to a certen extent;FBG of YXYZ and SJZTSZ were significantly decreased?p<0.01?,while other drug groups were decreased to some extent;The PRO of YXYZ and SJZTSZ were significantly decreased?p<0.05?,while the other drug groups were decreased to some extent;SCr of YXYZ significantly decreased?p<0.01?,SJZTSZ significantly decreased?p<0.05?,and other drug groups decreased to some extent;BUN of YXYZ significantly reduced?p<0.01?,while other drug groups to some extent reduced;Pathological sections of each drug group significantly intervented the diabetic renal tissue damage.ConclusionThe established quality control method has strong specificity and good repeatability,which can provide reference for the quality control of sijunzi decoction.Sijunzi decoction can regulate glucose-lipid metabolism disorder in T2DM mice,and ethanol extracts isbetter than water?0%ethanol?extracts.The Sijunzi decoction has certain intervention effect on diabetes-induced renal injury,could provide certain reference for the clinical intervention in diabetes-induced renal injury. |
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