利用培养组学技术分离鉴定体外降解胆固醇肠道细菌及其功能评价

Isolation and identification of cholesterol-degrading intestinal bacteria by culturomics and evaluation of their functions

摘要:

高胆固醇是诱发心脑血管疾病的重要因素之一。目前国内外降低胆固醇的主要方式是药物治疗，但其存在费用高及副作用多的弊端。研究表明，肠道细菌对机体的胆固醇代谢有重要影响，但降胆固醇肠道细菌的筛选方式及功能评价却少有报道。本研究通过培养组学方法，使用牛胆汁酸或人工混合胆汁酸作为筛选条件，从健康人体肠道筛选出36种耐胆汁酸细菌。以鼠李糖乳杆菌GG株 (Lactobacillus rhamnosus GG，LGG) 作为阳性对照，设置0 g/L、0.3 g/L、3 g/L三种胆汁酸浓度组，对耐胆汁酸细菌进行体外降胆固醇能力评估，确定奇异变形菌、斯氏普罗威登斯菌、普通变形菌等10种细菌为降胆固醇优势菌。随后对其中6种降胆固醇优势菌——奇异变形菌、斯氏普罗威登斯菌、普通变形菌、潘氏变形菌、污蝇解克杆菌、雷氏普罗威登斯菌进行体外降甘油三酯能力评估以及人工胃液耐受能力评估。结果显示，上述6株细菌体外降甘油三酯能力均优于LGG。伴随人工胃液pH值的下降和作用时间的延长，6株细菌的生存率均下降。上述筛选实验及功能评价为进一步开发潜在的降胆固醇细菌制品提供研究基础。

Abstract:

High cholesterol is one of the important factors inducing cardiovascular and cerebrovascular diseases. Drug therapy is the main method for reducing cholesterol, but has the disadvantages such as high cost and side effects. Studies have shown that intestinal bacteria play important roles in cholesterol metabolism. However, there are few reports on the screening and functional evaluation of cholesterol-lowering intestinal bacteria. In this study, 36 bile-tolerant bacteria were screened from healthy people stool through culturomics using bovine bile acid or artificial mixed bile acids as substrates. Taking Lactobacillus rhamnosus GG (LGG) as a positive control, three bile acid concentration groups (0 g/L, 0.3 g/L, 3 g/L) were set up to evaluate the cholesterol-lowering ability of bile-tolerant bacteria in vitro. Ten bacteria (including Proteus mirabilis, Providencia stuartii, Proteus vulgaris et al) were identified as the dominant cholesterol-lowering bacteria. Six of the above bacteria, Proteus mirabilis, Providencia stuartii, Proteus vulgaris, Proteus penneri, Wohlfahrtiimonas chitiniclastica, Providencia rettger, were evaluated for their ability to reduce triglycerides in vitro and tolerance to artificial gastric juice. Comparing with strain LGG, the six bacteria showed better triglyceride-lowering ability in vitro. With the decrease of pH value of artificial gastric juice and the increase of treatment time, the survival rate of six bacteria decreased. The above screening experiments and functional evaluation provide a basis for further development of potential cholesterol-lowering bacterial products.