Department of Biotechnology, Amity University Jharkhand, Ranchi, 834 002, India.
Department of Bio-Engineering, Birla Institute of Technology, Mesra, Ranchi, 835 215, India.
Indian J Gastroenterol. 2020 Oct;39(5):473-480. doi: 10.1007/s12664-020-01055-6. Epub 2020 Nov 17.
In our previous study, we have isolated different genera of bacteria from gallstones and this intrigued us to study their role in gallstone formation. The isolates exhibited certain biliary activities like urease activity, slime production, and β-glucuronidase production. We aimed to investigate the role of these factors in the formation of gallstone in in vitro conditions at a supersaturated concentration of cholesterol.
To mimic bile in in vitro state, Brilliant Green Bile Broth (BGBB) media having a composition similar to human bile was used. Four different experimental sets were prepared, each having nine flasks with varying concentrations of cholesterol and CaCO3 (calcium carbonate). Test sets I, II, III, and IV were inoculated with Salmonella, Enterococcus, Helicobacter, and Neisseria respectively, which were isolated from gallstone itself. Out of these four bacteria, only Helicobacter did not possess slime activity. A control set was also established which was devoid of bacteria. The control also had nine flasks with different concentrations of cholesterol and CaCO3. All the sets were incubated in the incubator shaker at 37 °C and 80 revolution per minutes (RPM) for 20 days.
It was observed that the sets having bacteria had a less nucleation time as compared to the control (F = 5.274; p < 0.001). Solidification of gallstone was observed only in the set with bacteria having slime activity (sets I, II, and IV).
The slime activity of bacteria leads to solidification of gallstones, whereas the other activities accelerate the nucleation of gallstone formation enhancing the severity of the disease.
在我们之前的研究中，我们从胆结石中分离出不同属的细菌，这引起了我们研究它们在胆结石形成中的作用。这些分离株表现出某些胆汁活性，如脲酶活性、黏液产生和β-葡萄糖醛酸酶产生。我们旨在研究这些因素在胆固醇过饱和浓度的体外条件下在胆结石形成中的作用。
为了模拟胆汁的体外状态，使用了组成类似于人胆汁的亮绿胆汁肉汤（BGBB）培养基。制备了四个不同的实验组，每个实验组有九个含有不同胆固醇和碳酸钙（碳酸钙）浓度的烧瓶。实验组 I、II、III 和 IV 分别接种了从胆结石本身分离出的沙门氏菌、肠球菌、幽门螺杆菌和奈瑟菌。这四种细菌中，只有幽门螺杆菌没有黏液活性。还建立了一个不含有细菌的对照实验组。对照组也有九个含有不同胆固醇和碳酸钙浓度的烧瓶。所有实验组都在 37°C 和 80 转/分钟（RPM）的摇床孵育器中孵育 20 天。
与对照组相比，发现含有细菌的实验组有更短的成核时间（F = 5.274；p < 0.001）。只有含有黏液活性的细菌的实验组（实验组 I、II 和 IV）观察到胆结石的固化。
细菌的黏液活性导致胆结石的固化，而其他活性则加速胆结石形成的成核，从而加重疾病的严重程度。
