粪便细菌移植对肝性脑病大鼠肝功能及血氨的影响

Effects of fecal microbiota transplantation on liver function and blood ammonia in hepatic encephalopathy rats

目的建立大鼠肝性脑病及FMT细菌模型,确定粪便细菌移植对肝性脑病大鼠肝功能及血氨的影响,初步了解粪便细菌移植(fecal microbiota transplantation,FMT)的临床意义。方法选择32只SD大鼠,采用CCL4+酒精的方法建立大鼠肝性脑病模型及肠道埋管,分为4组(A组:正常对照组;B组:肝性脑病模型组;C组:益生菌组;D组:FMT组),对C组进行益生菌移植,200μL/d(200μL益生菌溶液中含益生菌2.7×10~9),持续2周;对D组进行FMT移植,200μL/d(200μL粪便细菌溶液中含粪便细菌2.7×10~9),持续2周。记录大鼠一般生存情况及大便情况,对移植前后体重进行比较,测定移植前、后肝功能(ALT、AST、ALB、TB及DB)和门静脉及尾静脉血氨。结果所有实验大鼠均未出现死亡;粪便细菌移植后肝性脑病大鼠体重移植前明显增加[D组移植前(225.18±14.81)g,移植后2周(266.83±33.40)g,P<0.05];移植后大鼠肝功能较前明显好转[移植后2周ALT:D组与B组分别为(610.49±4.98)U/L和(990.07±4.80)U/L,P<0.05;AST:D组与B组分别为(72.46±4.42)U/L和(101.58±2.19)U/L,P<0.05;ALB:D组与B组比较ALB明显上升,分别为(24.48±0.12)g/L和(17.53±0.53)g/L,P<0.05;TB:D组为(13.45±0.77)μmol/L,B组为(27.20±0.77)μmol/L,P<0.05;DB:D组为(10.04±0.09)μmol/L,B组为(16.04±0.51)μmol/L,P<0.05];门静脉及尾静脉血氨较移植前明显降低[D组门静脉及尾静脉血氨分别为(26.26±0.30)μmol/L和(23.10±0.35)μmol/L,B组分别为(36.28±1.32)μmol/L和(32.90±0.35)μmol/L,P<0.05]。结论对大鼠进行人体粪便细菌移植是可行的;粪便细菌移植可改善肝性脑病大鼠的肝功能,降低血氨。

Objective To establish a rat model of hepatic encephalopathy （HE） and fecal microbiota transplantation （FMT） , investigate the effects of FMT on liver function and blood ammonia in order to explore the clinical significance of FMT. Methods Totally 32 rats were divided into 4 groups with 8 animals in each group, that is, normal control group, HE group, probiotics group, and FMT group. The SD rats model of HE was established through CCL4 ＋ alcohol injection, and the intestinal pipe was buried. The rats of the probiotics group were given probiotics transplantation （200 μL/d, containing probiotics of 2.7 × 10^9 ）, and those of the FMT group were given FMT （200 μL/d, containing fecal microbiota of 2.7 × 10^9 ）. After 2 weeks＇ treatment, the survival, defecation, and body weight, liver function （ALT, AST, ALB, TB and DB） and blood ammonia of portal vein and caudal vein before and after transplantation were recorded and compared. Results No death was observed in all experimental rats. When compared with the indexes before transplantation, FMT resulted in obvious increase in the body weight （266.83±33.40 vs 225.18 ±14.81 g,P 〈0.05 ） , significantly improved liver function （ FMT group vs HE group, ALT： 610.49 ±4.98 vs 990.07 ± 4.80 U/I., P 〈0.05; AST： 72.46 ±4.42 vs 101.58 ±2.19 U/L, P〈0. 05; ALB： 24.48 ±0.12 vs 17.53 ± 0. 53 g/L, P 〈0.05; TB： 13.45 ±0.77 rs 27.20 ±0.77 μmol/L, P 〈0.05; DB： 10.04±0.09 vs 16.04 ± 0.51 μmol/L, P 〈 0. 05 ）. The ammonia contents in the portal and tail venous blood were significantly decreased in the rats after than before FMT treatment （26.26 ±0.30 and 23. 10 ＋0.35 μmol/L respectively in the portal and tail venous blood in the FMT group, and 36.28 ± 1.32 and 32.90 ±0. 35 μ mol/L in the HE group, P 〈0.05）. Conclusion Human fecal bacteria transplantation is feasible in rats. FMT treatment can improve liver function and reduce the blood ammonia in HE rats.