| Objective: To observe the effect of heat-clearing turbid-expelling capsule on HOMA-IR,NO,level of ET-1,and expression of t-PA and PAI-1 in thoracic aorta in type-2 diabetic rats,to research the mechanism of its influence on islet function and action on macroangiopathy,so as to provide theoretical guidance on clinical treatment of type-2 diabetes and diabetic peripheral angiopathy.Methods: 40 healthy male adult Wistar rats weighing roughly 200 g were randomly sampled after 1-week adaptive feeding,8 of which were selected as blank controls by random number table,while the rest 32 were put in the modeling group.The rats in the normal control(NC)group were fed with basic forage,and administered intraperitoneally with normal saline at a time,the injection dose equaling the nicotinamide(NA)solution volume;the rats in the modeling group were fed by high-fat diet and administered intraperitoneally with low-dose streptozotocin(STZ)in order to build a T2 DM rat model.The rats were administered intraperitoneally with260mg/Kg NA solution,with 40mg/kg STZ(STZ dissolves in 0.1mmol/L in citric acid buffer,PH4.5)through caudal vein 15 min later,and with600000IU/kg VD3 solution in three days.Based on the above,the rats were fed by high-fat high-cholesterol diet(3% cholesterol,0.5% sodium cholate,0.2% propylthiouracil,5% sugar,10% lard and 81.3% basic forage)for 8weeks in a row.Meanwhile,the rats in all the treatment groups were administered intragastrically with different doses of different drugs.After modeling,the 32 rats were randomized into model group(8),Qingrequzhuocapsule group(8),Qingrequzhuo capsule + metformin group(8)and metformin group(8).The rats in the Qinrequzhuo capsule wereadministered intragastrically with(1.0g medicinal powder/kg/d),while the rats in the metformin control group were administered intragastrically with25 mg metformin/kg/d(the equivalent of 1500mg/d in adult people)once a day.The rats in the Qingrequzhuo capsule + metformin group were administered intragastrically with 25 mg metformin/kg/d(the equivalent of1500mg/d in adult people)and(1.0g medicinal powder/kg/d)once a day.The rats in the rest two groups were administered intragastrically with the same dose of purified water.8 weeks later,the rats fasted and underwent quick laparotomy after being administered intraperitoneally with 3% pentobarbital(80mg/kg)to be narcotized,from which the abdominal aorta was separated,from which blood was collected,and 2m L separated plasma was adopted for determination of ET-1,NO,insulin and CT peptide.When the abdominal aorta was taken,another piece was fixed by 4% paraformaldehyde and cut into slices with a thickness of 5mm after being washed,dehydrated,transparentized,waxed and encapsulated.The slices were tested by SABC method and dewaxed in a conventional way,with the endogenous peroxidase(EGPO)being blocked for 10 min in 3% H: 02,and washed for 3 times in distilled water,with the antigen retrieved by microwave;after the addition of confining liquid,rabbit antimouse PAI-1 and t-PA monoclone antibody were added;after a night at 4°C,Ig G was added on the next day,hatched for 20 min at 37°C and then SABC working solution was added.After color developing by DAB and counterstaining by hematoxylin,the slices were dehydrated,transparentized and encapsulated for testing.The color pathological image analysis system was used to calculate the positive staining area in the pathology slice tissues and the percentage of the window area.All measurement data was represented with X ±S,and paired t test was performed for an intra-group comparison;one-way ANOVA was conducted for intergroup comparisons,and Kruskal-Wallis H test was conducted for comparison among multiple independent samples in the case of variance non-homogeneity.All data was statistically processed by SPSS 17.0,and the test level ?=0.05.Results:1 Effects on the general state of rats: the rats in the normal control group were in good shape and energetic,drank water and took food as usual,and had snowy white hair,while the rats in the diabetes groups huddled together,frequently stayed still,and had a dull reaction,showing no statistical difference in water and food intake(P>0.05).Observation on the arterial histopathology in rats: Light microscope showed normal thoracic aortic tissues,closely junctional endocardial endothelial cells,narrow subendothelial spaces,densely and regularly arranged medial smooth muscle cells,light-colored plasma,and normal adventitial cells in the NC group,while pathological changes were not seen;irregular apophysis on endothelial cell surface,and a larger number of medial smooth muscle cells that invaded the intima,arranged in disorder and suffered structure disturbance in the DM group.Also,light microscope showed closely junctional endothelial cells,narrow subendothelial spaces,fewer proliferated medial smooth muscle cells,and significantly less-disordered cell arrangement in the Met,QRC and Met+QRC groups.There was no significant difference in arterial tissue structure between the three groups: both weight and urine volume increased,and hair color was dark.2 Effects on blood sugar: There was no statistical difference in blood sugar before modeling(P>0.05),compared with the NC group,the Met,QRC and Met+QRC groups saw a significant rise in blood sugar(P<0.05)before treatment,and there was no statistical difference in blood sugar between the DM group and the Met,QRC and Met+QRC groups(P>0.05).8 weeks later after intragastric administration,compared with the NC group,the Met,the QRC,QRC and Met+QRC groups saw a significant rise in blood sugar(P<0.05);there was no statistical difference in blood sugar between the Met+QRC group and Met group(P>0.05);there was a statistical difference in blood sugar between the DM group and Met group as well as Met+QRC group(P<0.05),so was between the QRC group and Met group as well as Met+QRC group(P<0.05);there was no statistical difference in blood sugarin the QRC group before and after treatment(P>0.05).There was a statistical difference in blood sugar in the Met and Met+QRC groups before and after treatment(P<0.05).3 Effects on HOMA-IR in rats: 8 weeks later after intragastric administration,compared with the NC,the Met,QRC and Met+QRC groups saw a significant rise in HOMA-IR(P<0.05).Compared with the DM group,the Met,QRC and Met+QRC groups saw a significant decline in HOMA-IR(P<0.05).There was no statistical difference in HOMA-IR between the Met group and Met+QRC group(P>0.05).Compared with the Met group,the QRC group saw a significant rise in HOMA-IR(P>0.05).4 Effects on the level of ET-1 in rats: 8 weeks later after intragastric administration,compared with the NC group,the Met,QRC and Met+QRC groups saw a significant rise in the level of ET-1(P<0.05).Compared with the DM group,the Met,QRC and Met+QRC groups saw a significant decline in the level of ET-1(P<0.05).There was no statistical difference in level of ET-1 between the Met group,QRC group and Met+QRC group(P>0.05).5 Effects on the level of NO: 8 weeks later after intragastric administration,compared with the NC group,the Met,QRC and Met+QRC groups saw a significant decline in the level of NO(P<0.05).Compared with the DM group,the Met,QRC and Met+QRC groups saw a significant rise in the level of NO(P<0.05).There was no statistical difference in level of NO between the Met group,QRC group and Met+QRC group(P>0.05).6 Effects on the expression of PAI-1 in thoracic aorta: Compared with the NC group where there was a little expression,the DM,Met,QRC and Met+QRC groups experienced a significant increase in expression in rat aorta(P<0.05).Compared with the DM group,the Met,QRC and Met+QRC groups experienced a significant decrease in expression(P<0.05).There was no statistical difference in expression between the Met group,QRC group and Met+QRC group(P>0.05).7 Effects on the expression of t-PA in thoracic aorta: Compared with the NC group where there was much expression,the DM,Met,QRC andMet+QRC groups experienced a significant decrease in expression in rat aorta(P<0.05).Compared with the DM group,the DM,Met,QRC and Met+QRC groups experienced a significant increase in expression(P<0.05).There was no statistical difference in expression between the Met group,QRC group and Met+QRC group(P>0.05).8 Observation on the arterial histopathology in rats: Light microscope showed normal thoracic aortic tissues,closely junctional endocardial endothelial cells,narrow subendothelial spaces,densely and regularly arranged medial smooth muscle cells,light-colored plasma,and normal adventitial cells in the NC group,while pathological changes were not seen;irregular apophysis on endothelial cell surface,and a larger number of medial smooth muscle cells that invaded the intima,arranged in disorder and suffered structure disturbance in the DM group.Also,light microscope showed closely junctional endothelial cells,narrow subendothelial spaces,fewer proliferated medial smooth muscle cells,and significantly less-disordered cell arrangement in the Met,QRC and Met+QRC groups.There was no difference between the Met group,QRC group and Met+QRC group.9 Drug side effects are compared: with NC group,Met group,QRC group,Met + QRC group rats all have mild diarrhea,diarrhea were similar among the three groups,did not see other reaction.compared with Met group,the Met + QRC group drug reaction is not increased.Conclusions:1 Compared with the DM group,the Qingrequzhuo capsule can reduce the HOMA-IR.2 The Qingrequzhuo capsule combined with metformin can reduce the level of ET-1 and increase the level of NO.3 The Qingrequzhuo capsule can inhibit PAI-1.4 Qingrequzhuo capsuleand promote the expression of t-PA in rat aorta.5 Qingrequzhuo capsule can Improve insulin resistance.6 This experiment suggests that the heat-clearing turbid-expelling capsule can and vascular endothelial diastolic and systolic dysfunction,adjustthe fibrinolytic system.7 Qingrequzhuo capsule +metformin Improve the insulin resistance effect is remarkable. |
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