The Effect Of Metformin On Intestinal Flora And Chronic Inflammatory State In Patients With Type2Diabetes Mellitus

Background&ObjectiveChronic Metabolic Diseases includes metabolic disorders and metabolism. It isa group of metabolic clinical syndrome, such as hypertension, diabetes mellitus,dyslipidemia, coronary heart disease, stroke and other diseases. Metabolic diseaseshas become a group of diseases that seriously threaten human health, under thebackground of genetic factors is relatively stable, environmental factors is particularlyimportant. With the development of microbial technology, more and more scientistsapply molecular microbiology in the research of intestinal flora. In human gut cavity,the colon has the most densely populated microorganism, different species, both thenumber and the species of bacteria is different in different lumen. Due to therestriction of technology and medical ethics, we can only through the research ofbacter in feces to understand intestinal flora indirectly. But whether the fecal flora canreflect mainly in the gut flora is not clear. Many studies will gut microbes to beinterpreted as environmental factors affect host nutrition, metabolism and immunefunction, to human body health is very important. The alteration of intestinal floraand low activity of chronic inflammatory state in the body may play an important rolein the development of diabetes and its complications.It has been paid much moreattention in recent years. Metformin as the first-line agent for patients with type2 diabetes the therapeutic efficacy get fully affirmed with English (UKPDS) results. Ofmetformin in patients with type2diabetes effect and mechanism of intestinal floraare reported. However, rarely research reports the metformin influence on thealteration of intestinal flora and low activity of chronic inflammatory state indiabetics. In this research, we aims to apply ERIC-PCR technology toestablishintestinal bacterial DNA fingerprint, preliminary discussion whether theMetformin can correct the alteration of intestinal flora and recrease the activity ofchronic inflammatory state in patients with T2DM.Subjects and Methods1． Subject collection The data are from patients who were diagnosised withtype2diabetes mellitus (T2DM) of the First Affiliated Hospital of ZhengzhouUniversity from February2013to June. Age from38to45years old,0.3～6years duration of diabetes. The diagnosis of the T2DM is correspond todiagnostic criteria made by Word Health Organization (WHO) in1999.2． Biochemical Collection The experimental group are given Melbine (0.85g,tid), regular application2weeks, collect fresh dejection of patients before andafter treatment, sealed in a sterile bio-bag; And extract peripheral blood beforeand after medication, take supernatant after4000rpm centrifuge for10minutes,all of the specimens are stockpile in a-80℃refrigerator less than half an hour.At the same time we observed the changes of blood glucose (FBG and2hBG),blood lipid and inflammatory factor (serum CRP、IL-6、and LPS).10healthy asa control.3． The Experiment Reagent and Instrument Intestinal flora genomic DNAextraction kit, Agarose, sterile PCR tubular; DNA marker, Dream Taq GreenPCR Master Mix, Green DNA-Dye and nucleotides colloidal Dye and primers.4． Experiment Steps1) To extracted the DNA of Intestinal flora, washing themanure with saline after freeze-thaw, taking300g of sediment into thesterilization EP tube after4000rpm centrifuge for1minute, then according tothe instruction to extract DNA.2) To amplified it by ERIC-PCR fingerprint analysis technique for amplification, design the specific primers as ERIC1(5’-ATG AAG CTC CTG GGG AAT CCA a3’), ERIC2(5’-AAG TAA GTG ACTGGG GTG ATG CG a3’).3) The PCR product was subjected to electrophoresisand the results were analyzed by gel imaging and analysis system.4) fingerprintatlas analysis5) Using Elisa kit to detection the serum inflammatory factors.6)Fasting and postprandial blood glucose was determined by glucose oxidase,biochemiacal indexes were measure in the clinical lab of our hospital.5． Statistical Methods SPSS17.0statistical software was used to analyze data.All continuous variables of the normal distribution were showed withmean±standard deviation (X±S). After homogeneity of variance test andnormality test, independent sample t test was used to compare two arrays of data.If P<0.05, there’s statistical significanc.Results1． After the drug was administered the lever of FBG、2hBG and serium TC wassignificantly lower(P＜0.05); serium TG、LDL level was lower, and the HDLwas risen, but there were no statistically significant difference (P＞0.05).Aftermedication, serium CRP、LPS、IL-6level was significantly l lower(P＜0.05)。2． After medication, serium CRP、LPS、IL-6level was significantly l lower(P＜0.05).3． ERIC-PCR fingerprinting map shows the number of DNA banding in healthyintestinal flora is more than in patients with T2DM. The brighter band isdifferent, and individual DNA lighter stripe is scattered distribution. The numberof DNA bands in intestinal flora in patients with T2DM increased after takingmetformin, and the lighter stripe is different, similar to healthy people.Conclusion1． After the drug was administered the seriun lever of FBG、2hBG、TC、CRP、TNF-a、LPS was significantly lower. It suggests that metformin helps reduceinflammatory factor levels, improve the state of patients with chronic inflammation, improved blood sugar and lipid levels. While some study showedthat the anti-inflammatory action of metformin is independent of hypoglycemiceffect.2． Through the analysis of the intestinal flora, the kind of health intestinal flora isabundant in healthy individuals, the dominant bacteria has a large number kinds.The structure of intestinal flora diversity of patients with type2diabetes isreduced, and the dominant bacteria is different from health one. After medication,the structure of intestinal flora of patients with type2diabetes was changed, theyare similar to those of health people. In this study, we can only observe thechange tendency of intestinal flora after be given menformin. However type andquantity of specific changes of intestinal flora needs to be studied further.