[논문]Enterococcus faecium R0026 Combined with Bacillus subtilis R0179 Prevent Obesity-Associated Hyperlipidemia and Modulate Gut Microbiota in C57BL/6 Mice

Huang, Jinli; Huang, Juan; Yin, Tianyi; Lv, Huiyun; Zhang, Pengyu; Li, Huajun

doi:10.4014/jmb.2009.09005

Enterococcus faecium R0026 Combined with Bacillus subtilis R0179 Prevent Obesity-Associated Hyperlipidemia and Modulate Gut Microbiota in C57BL/6 Mice 원문보기

Journal of microbiology and biotechnology, v.31 no.2, 2021년, pp.181 - 188

Huang, Jinli (Department of Microecology, College of Basic Medical Sciences, Dalian Medical University) , Huang, Juan (Department of Microecology, College of Basic Medical Sciences, Dalian Medical University) , Yin, Tianyi (First Affiliated Hospital of Dalian Medical University) , Lv, Huiyun (First Affiliated Hospital of Dalian Medical University) , Zhang, Pengyu (Department of Microecology, College of Basic Medical Sciences, Dalian Medical University) , Li, Huajun (Department of Microecology, College of Basic Medical Sciences, Dalian Medical University)

Abstract ▼ AI-Helper

Bacillus subtilis and Enterococcus faecium are commonly used probiotics. This study aimed to identify the effect of live combined Bacillus subtilis R0179 and Enterococcus faecium R0026 (LCBE) on obesity-associated hyperlipidemia and gut microbiota in C57BL/6 mice. Forty male C57BL/6 mice were divided into four groups: normal group (N group), model group (M group), low-dose group (L group), and high-dose group (H group). Mice were gavaged with LCBE at 0.023 g/mice/day (L group) or 0.23 g/mice/day (H group) and fed with a high-fat diet for 8 weeks. In vitro E. faecium R0026 showed an ability to lower the low-concentration of cholesterol by 46%, and the ability to lower the high-concentration of cholesterol by 58%. LCBE significantly reduced the body weight gain, Lee index, brown fat index and body mass index of mice on a high-fat diet. Moreover, LCBE markedly improved serum lipids (including serum triglyceride, total cholesterol, low-density lipoprotein and high-density lipoprotein) while also significantly reducing liver total cholesterol. Serum lipopolysaccharide and total bile acid in L and H groups decreased significantly compared with M group. PCR-DGGE analysis showed that the composition of gut microbiota in the treatment groups was improved. Akkermansia muciniphila was found in H group. The PCA result indicated a similar gut microbiota structure between LCBE treatment groups and normal group while the number of bands and Shannon diversity index increased significantly in the LCBE treatment groups. Finally, qPCR showed Bifidobacterium spp. increased significantly in H group compared with M group, LCBE alleviated liver steatosis and improved brown adipose tissue index.

주제어

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문제 정의

This study aimed to evaluate the influence of LCBE on obesity-associated hyperlipidaemia and gut microbiota in C₅₇BL/6 mice.

제안 방법

CN₁₀4371958 A). LCBE orally or alone (1.0×10⁸ CFU/kg/day) for 3 days was able to normalize the Treg/CD4+ ratio in the blood and mesenteric lymph nodes in a female Wistar rat model. LCBE also normalized the smooth muscle contractility in post inflammation rats [20].
ACC GCG GCT GG-3') targeting the V3 region. PCRs were conducted in a total volume of 50 μl systems, 25 μl of Master Mix, 1 μM 338f and 518r primers, and 20 μl of RNase-free water. Three microliters of DNA was used as a template.

대상 데이터

B. subtilis R0179 and E. faecium R0026 were from probiotic product Medilac-S (Hanmei Pharmaceuticals, China), which was composed of 4.4×10⁹ colony-forming units (CFU) of B. subtilis R0179 and 6.0 × 10¹⁰ CFU of E.faecium R0026.
Forty male C₅₇BL/6 mice (four-week-old) were bought from Dalian Medical University. Mice were housed five individuals per cage and bred in pairs in an SPF laboratory animal house (Model Animal Research Centre, Dalian Medical University, China).

데이터처리

The real-time PCR data were analyzed by MxPro-Mx3005P System and software (Agilent, USA). The unpaired Student’s t-test was used to compare two groups and one-way analysis of variance was used to compare the varieties between all groups. A statistically significant probability value was p < 0.

이론/모형

The relative genes expression of gut microbiota normalized to 16S rRNA expression and internal reference in fecal samples using the comparative CT (threshold cycle) method [33]. All results were normalized and calculated by using the 2^−ΔΔCT method.
Next, a visible light homogeneous phase removal method based on the selective action of mixed surfactants and a chemiluminescence homogeneous phase removal method were used to determine serum HDL and LDL [25]. Serum total bile acid (TBA) was measured by the enzymatic colorimetric method. (TBA concentration = ΔA determination/ΔA standard × standard concentration (μmol/l)) [26].
The specific 16S rRNA primers amplifying genes melting curve analysis of products was used to confirm that fluorescence signal originated from the specific amplification product. The relative genes expression of gut microbiota normalized to 16S rRNA expression and internal reference in fecal samples using the comparative CT (threshold cycle) method [33]. All results were normalized and calculated by using the 2^−ΔΔCT method.

성능/효과

LCBE modulated gut microbiota in rats with ALD [40]. In this study, parameters related to dyslipidaemia (TC, TG, LDL) were reduced by LCBE. High-dose LCBE significantly improved the HDL.
Our results demonstrated that live Bacillus subtilis R0179 and Enterococcus faecalis R0026 reversed obesity in mice. There is a great interest for identifying novel probiotics with anti-obesity function.

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표제어: PCR

동의어: Packet Collision Rate

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