灭活的双歧杆菌抗肠源性感染的实验研究

发布时间：2018-03-29 10:09

本文选题：双歧杆菌　切入点：菌群失调　出处：《延边大学》2007年硕士论文

【摘要】： 双歧杆菌作为人和动物肠道中最重要的生理性细菌之一，对机体无毒、无害、并且具有广泛的生态效应与生理作用。双歧杆菌参与了宿主的消化、营养、代谢、吸收、免疫及抗感染过程，被认为是微生态学研究的核心或重心。现有研究表明，双歧杆菌制剂可恢复肠道内菌群失调的生态平衡，保护并修复肠道黏膜，促进肠道益生菌增殖，因而在防治肠源性感染中起到了良好的治疗效果。目前国内外研制开发的双歧杆菌制剂采用的都是活菌，其有效期不过一年左右，这样就限制了此类产品的应用范围与条件。如果我们将灭活的双歧杆菌制成微生态调节剂，进行工业化生产，通过提高产品质量来增强其竞争力，，就可以打破国外此类产品对国内市场的垄断，因此本研究对推动微生态学基础理论研究有着重要意义。本实验将双歧杆菌接种至BS肉汤培养基中，37℃厌氧培养48h。所得菌悬液分为活菌组和死菌组。将活、死两组菌悬液用NS洗3次，离心弃上清，用SCS重悬活、死双歧杆菌沉淀，使菌悬液浓度均达到1×10~(11) CFU／mL。将昆明种小鼠随机分为活菌组、死菌组、自然恢复组、正常对照组4组，每组10只。除正常对照组外，其余3组小鼠分别造成抗生素致菌群失调、环磷酰胺致免疫功能低下及烧伤动物模型。从第4d开始，活菌组采用双歧杆菌活菌悬液灌胃；死菌组采用双歧杆菌灭活菌悬液灌胃；自然恢复组采用NS灌胃，连续3d。4组小鼠分别摘眼球采血处死，利用日本OLYPUS-AU400全自动生化仪对临床血清学指标进行测定。实验结果如下： 1．灭活的双歧杆菌治疗小鼠菌群失调致肠源性感染采用腹腔注射青霉素造成抗生素菌群失调小鼠动物模型，分别以双歧杆菌灭活菌液、活菌液对小鼠进行灌胃治疗后，对血清总蛋白(TP)、血清白蛋白(ALB)、血清球蛋白(GLO)、y-谷氨酰氨转肽酶(GGT)、血清尿素氮(BUN)、乳酸脱氢酶(LDH)、血肌酐(CRE)以及组织匀浆内的示踪菌进行测定。结果显示，死菌组与自然恢复组比较，TP、GLO明显升高，GGT、BUN则显著降低。死菌组与活菌组、正常对照组比较，LDH明显降低，死菌组与自然恢复组比较，肾、肝、肺组织匀浆内肠杆菌数量均明显下降。 2．灭活的双歧杆菌治疗免疫功能低下致小鼠肠源性感染采用腹腔注射环磷酰胺造成免疫功能低下小鼠动物模型，分别以双歧杆菌灭活菌液、活菌液对小鼠进行灌胃治疗后，检测临床血清学指标与组织匀浆内示踪菌。结果显示，活菌组、死菌组同自然恢复组相比，临床血清学指标水平与组织匀浆内示踪菌数量恢复至正常对照组水平。 3．灭活的双歧杆菌治疗烧伤致小鼠肠源性感染制作Ⅲ度烫伤(烧伤)小鼠动物模型，分别以双歧杆菌灭活菌液、活菌液对小鼠进行灌饲后，检测组织匀浆内示踪菌数量与血清蛋白水平。结果显示，活菌组、死菌组同自然恢复组相比，组织匀浆内示踪菌数量与血清蛋白水平恢复至正常对照组水平。结论： 1．双歧杆菌灭活菌悬液具有与活菌悬液相同的抗菌群失调致小鼠肠源性感染作用。 2．双歧杆菌灭活菌液具有与活菌液相同的抗免疫功能低下致小鼠肠源性感染作用。 3．双歧杆菌灭活菌液具有与活菌液相同的治疗小鼠烧伤感染作用。
[Abstract]:Bifidobacterium as one of the most important physiological bacteria in human and animal intestinal tract, harmless to the body, non-toxic, and has ecological effects and extensive physiological functions. Bifidobacteria participate in host digestion, nutrition, metabolism, absorption, immunity and anti infection, is considered to be the core of micro ecology research or existing focus. Studies show that bifidobacteria can restore intestinal flora ecological balance, protection and repair of the intestinal mucosa, promote the proliferation of probiotics, which played a good therapeutic effect in the prevention of enterogenic infection. At home and abroad developed by Bifidobacterium are viable, the period of validity but a year or so, thus limiting the application scope and conditions of such products. If we made the inactivated Bifidobacterium micro ecological regulator, industrial production, to increase by improving the quality of products Strong competitiveness can break the monopoly of foreign products to the domestic market. Therefore, this study is of great significance to the research of the basic theory of microecology.
The bifidobacteria were inoculated in BS broth, 37 C anaerobic bacteria suspension culture of 48h. income divided into viable group and death group. The live bacteria, dead bacteria suspension with two were washed 3 times with NS, centrifuged supernatant, with SCS suspending live Bifidobacterium bacteria to death precipitation. The suspension concentration reached 1 * 10~ (11) CFU / mL. Kunming mice were randomly divided into group of live bacteria, dead bacteria group, natural recovery group, normal control group 4 groups, 10 rats in each group. Except the normal control group, the remaining 3 groups of mice were caused by antibiotic induced dysbacteriosis, cyclophosphamide induced immune hypofunction and burn animal model. From the beginning of 4D, bacteria group with Bifidobacterium suspension gavage; dead bacteria group using Bifidobacterium inactivated bacteria suspension gavage; natural recovery group gavaged with NS, continuous 3d.4 mice were eyeball blood were killed, the clinical blood indexes using OLYPUS-AU400 automatic biochemical analyzer Determination.
The experimental results are as follows:
1. inactivated bifidobacteria in the treatment of intestinal dysbacteria induced intestinal infection in mice
By intraperitoneal injection of penicillin antibiotics dysbacteriosis caused by the animal model of mice were Bifidobacterium inactivated and live bacteria into the stomach of the mice after treatment, the serum total protein (TP), serum albumin (ALB), serum globulin (GLO), y- glutamine transpeptidase (GGT), serum urea nitrogen (BUN), lactate dehydrogenase (LDH), serum creatinine (CRE) and tissue homogenate in tracer bacteria were determined. The results show that the death group, bacteria group and natural recovery of TP, GGT, GLO increased significantly, BUN decreased significantly. The dead bacteria group and normal bacteria group. Compared to the control group, LDH decreased significantly, the death group, bacteria group and natural recovery of kidney, liver and lung homogenate in enterobacteria decreased significantly.
2. inactivated bifidobacteria in the treatment of intestinal infection in mice induced by immunodeficiency
By intraperitoneal injection of cyclophosphamide caused by low immune function mice animal model, respectively Bifidobacterium inactivated and live bacteria into the stomach of the mice after treatment, detection of clinical serum markers and tracer bacteria within tissue homogenates. The results showed that the living bacteria group, dead bacteria group and natural recovery group compared to the level of clinical and serological indicators the number of bacteria in the tissue homogenate tracer recovered to the normal level of the control group.
3. inactivated Bifidobacterium for the treatment of intestinal infection in mice caused by burn
Production of scald (burn) animal model of mice were Bifidobacterium inactivated and live bacteria liquid on mice after intragastric administration, the number of bacteria detection tracer homogenate and serum protein level. The results showed that the living bacteria group, dead bacteria group and natural recovery group compared to tissue homogenate of cytoplasm of tracer bacteria the number and serum protein levels returned to the normal level of the control group.
Conclusion:
1. the suspension of Bifidobacterium inactivated bacteria has the effect of intestinal infection in mice caused by the same antibacterial group as the living bacteria suspension.
2. Bacillus Bifidobacterium inactivated bacteria has the same anti immune function as the living bacteria, which induces intestinal infection in mice.
3. Bifidobacterium inactivated bacteria have the same effect as living bacteria solution in the treatment of burn infection in mice.

【学位授予单位】：延边大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R371

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