| A variety of microorganisms which are referred to intestinal flora live in the gastrointestinal tract of people and animals. Among these microorganisms, bacteria own the largest proportion. The intestinal flora impact on the human health and flora disorders in the host could cause many diseases, such as type2diabetes, enteritis and chronic hepatitis. Type2diabetes is a kind of endocrine and metabolic diseases in pathophysiological basis of insulin resistance and insulin secretion. Recent studies have shown that intestinal bacteria is closely associated with insulin resistance, which play an important role in the development ans long-term energy homeostasis in type2diabetes. The combination of Radix Scutellariae and Rhizoma Coptidis, so-called Scutellaria-Coptis herb couple is widely used for detoxification in clinic especially in treating type2diabetes. Main active ingredients of Radix Scutellariae are flavonoids, such as baicalin, baicalein, wogonoside and scutellarin; main active components of Rhizoma Coptidis are alkaloid, such as berberine, coptisine and palmatine. Most traditional medicines are oral adiminstrated and then exert their therapeutic effects. On the one hand, a large number of intestinal bacteria can produce abundant enzymes, resulting in the active ingredient be converted into a series of metabolites absorbed into the blood; the other hand, some of the active ingredients inhibite the growth of harmful bacteria and promote the growth of beneficial bacteria, so as to achieve the purpose of adjusting intestinal flora.Thus, the study on interactions between scutellaria-coptis herb couple and intestinal bacteria is helpful for fully understanding the mechanism of absorption, metabolism and action of scutellaria-coptis in vivo. In addition, the study also provides a preliminary theory for revealing the treatment of type2diabetes in microbiology mechanism.1Scutellaria-Coptis metabolism by human intestinal bacteriaDifferent types of bacterial colonies have been isolated from human feces. Baicalin, wogonoside, scutellarin, berberine copstisine, palmatine, Radix Scutellariae extracts, Rhizoma Coptidis extracts and Scutellaria-Coptis extracts were incubated under anaerobic condition with the intestinal mixture and the isolated bacteria, respectively. The intestinal bacteria, capable of metabolizing above compounds were screened to identify by16S rRNA. Additionally, an ultra performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UPLC/QTOF-MS) technique combined with MetabolynxTM software was established and applied for the detection and identification of the metabolites by the isolated human intestinal bacteria.Human intestinal bacteria capable of metabolizing Scutellaria-Coptis were classified to Escherichia, Bacillus and Enterococcu. About80%of the bacteria owned the demethoxy function, but a small amount of berberine, coptisine and palmatine could be metabolized by them and the types and contents of products were quite less. Nearly half of the bacteria got the function of deglycosylation, which aimed to flavonoids (baicalin, wogonoside and scutellarin). Aglycones were the dorminant products after deglycosylation. About30%-40%of the bacteria owned the function of methylation. Reducion function of intestinal bacteria in this experiment was in the performance of hudrogenation and the number of the bacteria was about20%-30%. Additionally, a small portion of bacteria, about10%owned the function of ring cleavage and demetylation. The same drugs were transformed by mixed bacteria and a single strain, the metabolic capacity of mixed bacteria was weaker relatively. After single herb compatibility, the type and content of products in mixed bacteria were reduced.2Interactions between rat intestinal bacteria and Scutellaria-CoptisA model of type2diabetes rat was established with the method of feeding high-fat diet combined with STZ injection. Radix Scutellariae extracts, Rhizoma Coptidis extracts and Scutellaria-Coptis extracts were incubated under anaerobic condition with normal rat intestinal mixture. Based on the above-mentioned monomer components, the metabolites were screened and identified. Apart from this, two types of beneficial bacteria (Bifidobacteria sp., Lactobacilli sp.) and two types of harmful bacteria (Enterococcous sp., Enterobacter sp.) were isolated from normal and type2intestinal bacteria respectively. The effects of Radix Scutellariae extracts, Rhizoma Coptidis extracts and Scutellaria-Coptis extracts on the growth of bacteria were detected by enzyme-labeled instrument.Normal rat intestinal bacteria metabolized main components of Radix Scutellariae to cleavage products, deglycosylated products, deglycosylatedand methylated products, hydrogenated products, methylated products; the ability of metabolizing alkaloids in Rhizoma Coptidis was weak. Abilities of intestinal bacteria to metabolize herb couple were relatively weak. The products of Radix Scutellariae were cleavage product, deglycosylated and methylated products, deglycosylated product and methylated products; while alkaloids in Rhizoma Coptidis were metabolized hardly. Deglycosylation was the main metabolic way of type2diabetic rat intestinal bacterial and the content of baicalein increased significantly. While the ability of type2diabetic rat intestinal bacterial metabolizing alkaloids was still quite week. The data of drug effects on bacteria growth showed that Radix Scutellariae extracts, Rhizoma Coptidis extracts and Scutellaria-Coptis extracts all stimulated the growth of beneficial bacteria and inhibited the growth of harmful bacteria. Furthermore, the ability of Scutellaria-Coptis extracts inhibiting the harmful bacteria growth was stronger. 3Study on Scutellaria-Coptis in vivoSome metabolic difference happened to type2diabetic rat intestinal bacteria lead to different absorption and metabolism in vivo. Baicalin, baicalein, wogonin, berberine, coptisine and palmatine were selected as detection index. We established a method of simultaneous determination of these six ingredients rat plasma by UPLC-MS/MS and applied to the pharmacokinetic comparative study in the rats after intragastric administration of Radix Scutellariae, Rhizoma Coptidis and their herb couple and provide some scientific basis of the compatibility of Scutellaria-Coptis herb couple. UPLC-Q-TOF/MS and MetaboLynx software based on mass defect filter (MDF) were applied to analyze the metabolites in the urine, plasma, bile and feces of normal rats after intragastric administration of Radix Scutellariae, Rhizoma Coptidis and their herb couple. Additionally, studies on the Caco-2cellular uptake properties of baicalin, baicalein, wogonin, berberine, coptisine and palmatine were of great importance for investigating the mechanism of drug absorption and improving clinical therapy clinical efficacy.Pharmacokinetic parameters displayed that the bioavailability of baicalin and berberine in type2diabetic rats significantly increased compared with normal rats. Metabolism data in vivo showed that baicalin and berberine in type2diabetic rat blood levels were more than those in normal samples. Data of Caco-2cell model showed that the permeability coefficient increased with the substrate concentration increased and decreased with time; glycosides were more difficult to absorb than aglycones; ingredients’permeabilities in a single herb was significantly higher than that in herb couples.The metabolism of Scutellaria-Coptis herb couple by intestinal bacteria was related with type2diabetes. One hand, drugs were metabolized by intestinal bacteria to easily absorbed components and these components were converted in vivo to the therapeutically effective substances; the therapeutic substance metabolism could be metabolized by bacteria and absorbed into the blood directly. On the other hand, drugs contact with gastrointestinal tract intestinal microflora after oral administration and supported beneficial bacteria and inhibited harmful bacteria, which could reduce the number of inflammatory cytokines in the blood and delaying the onset of diabetes complications. Thus, the research about the metabolism of Scutellaria-Coptis herb couple by normal and type2diabetic rat intestinal bacteria is helpful for fully understanding the mechanism of absorption, metabolism and action of this compound in vivo, guiding clinical medication and investigating new drug. |
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