Effects Of Berberine Hydrochloride On The Intestinal Flora And Serum Endotoxin Level In Rats With T2DM

Objective:Objective to observed the relation between serum endotoxin level and Insulin resistance in type 2 diabetes mellitus(T2DM) rats of the processing, to observe the effect of berberine hydrochlorideand on the intestinal flora、serum endotoxin level in T2 DM rats, to investigate the mechanism of berberine hydrochloride treating T2 DM rats.Methods:Sixty-six male SD rats were randomly divided into 2 groups(ages of 6 to 8 weeks,Weight is 180 g to 200 g): group NC(sham operated group, n=24), group T2DM(T2DM group, n=42). In group NC received ordinary feed. T2 DM group rats fed by the high sugar,high fat diet after 12 weeks, the overnight fast 8 hours give intraperitoneal injection of streptozotocin(STZ) 30 mg/kg, after every 2 days from the rat tail take rats fasting blood glucose(FBG), blood test tendency for more than 2 consecutive FBG > 7.8mmol/ L,namely that a building success. Feed formulation of the high sugar, high fat diet:basal feed65%, lard 10%, sucrose 15%, Dried egg yolk 10%.Randomly select NC group and T2 DM group rats all six only in the four weeks, 8weeks and after the construction of the model, these rats received intraperitoneal injection of 4ml/kg 20% urethane after 8 hours fasting. These rats were measured weight, sampled abdominal aortic blood, separated the plasma and serum. To return part of the right liverissue, physiological saline flushing after quickly into the embedding box, made into 10%formalin in the preparation of wax block dewater fixed after 24 hours.T2DM group success in the T2 DM rats 20, the 18 similar weight rats were randomly divided into High sugar and high fat diet group(HSHF group, n=6,continuing fed by the high sugar, high fat diet),Simple diet treatment group(group Ⅰ, n=6, received ordinary feed), berberine with diet therapy group(group Ⅱ, n=6, received ordinary feed and lavaged berberine hydrochloride 200mg/kg/d),. After 8 weeks, handle with sham operated group rats, these rats were measured weight, sampled abdominal aortic blood, separated the plasma and serum. To return part of the right liver issue, physiological saline flushing after quickly into the embedding box, made into 10% formalin in the preparation of wax block dewater fixed after 24 hours. To return a small amount of colon in feces, one part of fixed immediately smear, another part of the 2ml inserted in RNase EP liquid nitrogen frozen moved over 30 minutes- 80℃ refrigerator.The expression of serum endotoxin was examined by the limulus reagent chromogenic substrate method. The expression of C-reactive protein(CRP),cereal third transaminase(ALT), triglycerides(TG), fasting blood glucose(FBG) were examined by immune turbidity method. The expression of fasting serum lisulin(FINS)was examined by radioimmunoassay. Calculated the insulin resistance index(HOMA-IR),The formula: FPG(mmol/L)×FINs(m IU/L)/22.5. The proportion of cocci and coli of intestinal flora were caculated by microscope. The number of bifidobacterium, lactic acid bacteria, E. coli in intestinal flora was examined by RT-PCR. Liver wax block line of HE staining to observe the liver tissue pathological changes.Results:1.The relationship between the level of insulin resistance and the level of serumendotoxin in the course of T2 DM formation induced by high gluoose and high-fat diet combined with STZ in rats.1.1 In the fourth weekend. The levels of rats weight, ALT, TG, CRP, LPS in group T2 DM were more than that in group NC(P<0.05). There were no differences in the levels of FPG, FINS, HOMA-IR between group T2 DM and group NC. Liver tissue pathological observation in group T2DM: vesicles lipid drops of liver steatosis, ballooning degeneration.1.2 In the eigth weekend, The levels of rats weight, ALT、TG、CRP、LPS、FPG、FINS、HOMA-IR in group T2 DM were more than that in group NC(P<0.05).Liver tissue pathological observation in group T2DM: the big bubble lipid drops of liver steatosis, ballooning degeneration.1.3 The T2 DM rats’ s model establishment. Compared with group NC, the rats’ s colour lackluster, Less activity. The levels of rats weight, ALT、TG、CRP、LPS、FPG、FINS、HOMA-IR in group T2 DM were more than that in group NC(P<0.05).Liver tissue pathological observation in group T2DM: diffuse cavitation fatty degeneration,steatosis, ballooning degeneration, inflammatory cells infiltration, aggravating the degree of inflammation.1.4 The level of LPS was positively correlated with the degree of IR,r=0.740,p<0.001.1.Berberine and Simple diet treatment of T2 DM the possible mechanism:2.1 Compared with group NC,the levels of rats weight, ALT、TG、CRP、LPS、FPG、HOMA-IR in groupⅠ、in groupⅡwere more than that in NC(P<0.05).The lesion of steatosis, ballooning degeneration, inflammatory cells infiltration were lingt than that in groupⅢ.2.2Compared with group HSHF, the levels of rats weight, ALT、TG、CRP、LPS、FPG、FINS、HOMA-IR in groupⅠ、in groupⅡwere less than that in group HSHF(P<0.05). Liver tissue pathological observation in group HSHF: steatosis,VIII ballooning degeneration, inflammatory cells infiltration, dot focal necrosis.2.3 The change of intestinal microflora after T2 DM rats treated with Berberine and simple diet:2.3.1Berberine hydrochloride, simple diet therapy in T2 DM rats after the gram negative bacterium Escherichia coli belongs to the change of NC group compared to berberine hydrochloride group, simple diet group of Escherichia coli is abundance decreased(P < 0.05). Compared with group HSHF, berberine hydrochloride group, simple diet group of Escherichia coli is abundance decreased(P < 0.05); compared with diet alone group and hydrochloric acid berberine group of Escherichia coli is significantly reduced(P< 0.05).2.3.2Berberine hydrochloride, simple diet in the treatment of type 2 diabetic rats after the probiotic Lactobacillus and Bifidobacterium species changes with NC group compared to the berberine hydrochloride group, simple diet group of lactic acid bacteria,Bifidobacterium is abundance increased(P < 0.05); compared with group HSHF, berberine hydrochloride group, simple diet group of lactic acid bacteria, Bifidobacterium genus abundance increased(P < 0.05); compared with diet alone group, hydrochloride berberine group of lactic acid bacteria, Bifidobacterium genera abundances increased significantly(P< 0.05).Conclusions:(1) High glucose and high fat diet caused mild elevation of LPS in rats, and induced chronic low-grade inflammation in rats, and to IR, after STZ injection, insulin secretion was relatively insufficient, T2 DM produced.(2) Berberine hydrochloride in the treatment of T2 DM rats and its possible mechanism is through improving the structure of intestinal bacteria group, reduce bloodlevels of LPS into the body, thereby easing endotoxemia, achieve the purpose of therapy in the treatment of T2 DM.