肝硬化合并腹水患者的发病危险因素分析

Investigation of risk factors and application analysis of 16S rDNA in patients with partial cirrhosis combined with ascites

目的探究肝硬化合并腹水患者的发病危险因素,为肝硬化合并腹水的防治提供依据。方法选取健康人68名(健康对照组)、肝硬化代偿期患者70例(肝硬化代偿组)和肝硬化失代偿期患者70例(肝硬化失代偿组)。采用酶联免疫吸附测定法(ELISA)试剂盒检测各组研究对象肝功能指标,采用流式血细胞分析仪检测各组研究对象白细胞计数(WBC)、中性粒细胞计数(NE)、血小板计数(PLT),采用16S rDNA分析各组研究对象肠道菌群情况,采用ELISA试剂盒测定各组研究对象炎症因子及内毒素水平,采用蛋白印记法检测各组研究对象腹水相关蛋白表达水平,对上述指标进行多因素Logistic回归分析,分析部分肝硬化合并腹水患者的发病危险因素。结果与健康对照组比较,肝硬化代偿组患者肠杆菌和肠球菌计数较高,双歧杆菌和乳杆菌计数较低(P<0.05);与肝硬化代偿组比较,肝硬化失代偿组患者肠杆菌和肠球菌计数较高,双歧杆菌和乳杆菌计数较低(P<0.05);与健康对照组比较,肝硬化代偿组患者丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)、总胆红素(TBIL)较高,白蛋白(ALB)水平较低(P<0.05);与肝硬化代偿组比较,肝硬化失代偿组患者ALT、AST和TBIL水平较高,ALB水平较低(P<0.05);与健康对照组比较,肝硬化代偿组患者WBC、PLT和NE水平较低(P<0.05);与肝硬化代偿组比较,肝硬化失代偿组患者WBC、PLT、NE水平较低(P<0.05);与健康对照组比较,肝硬化代偿组患者透明质酸(HA)和Ⅲ型前胶原肽(PⅢP)水平较高(P<0.05);与肝硬化代偿组比较,肝硬化失代偿组患者HA和PⅢP水平较高(P<0.05);与健康对照组比较,肝硬化代偿组患者白细胞介素6(IL-6)、白细胞介素8(IL-8)、肿瘤坏死因子α(TNF-α)和脂多糖(LPS)水平较高(P<0.05);与肝硬化代偿组比较,肝硬化失代偿组患者IL-6、IL-8、TNF-α、LPS水平、Toll样受体4(TLR4)、核因子κB(NF-κB)、c-Jun氨基末端激酶(JNK)、p38丝裂原活化蛋白激酶(p38 MAPK)表达水平均升高(P<0.05)。肠杆菌、肠球菌、双歧杆菌、乳杆菌、LPS和TLR4是肝硬化合并腹水的危险因素。结论肝硬化患者临床特征与肠道菌群存在一定的关联,其中炎性、免疫等指标、肠道菌群均与肝硬化腹水的发生发展密切相关,肠杆菌、肠球菌、双歧杆菌、乳杆菌、LPS和TLR4是肝硬化合并腹水的危险因素。

Objective To investigate the risk factors for cirrhosis complicated by ascites and provide a basis for the prevention and treatment of cirrhosis with ascites.Methods A total of 68 healthy individuals(healthy control group),70 patients with compensated cirrhosis(compensated cirrhosis group),and 70 patients with decompensated cirrhosis(decompensated cirrhosis group) were selected.Liver function indicators of each group were detected using enzyme-linked immunosorbent assay(ELISA) kits.White blood cell count(WBC),neutrophil count(NE),and platelet count(PLT) were detected using a flow cytometer.The intestinal flora of different groups was analyzed using 16S rDNA analysis.Inflammatory factors and endotoxin levels were measured using ELISA kits.The expression levels of ascites-related proteins were detected using western blotting.Multivariate logistic regression analysis was performed on these indicators to identify the risk factors for cirrhosis with ascites.Results Compared with healthy control group, the counts of Enterobacter and Enterococcus were higher, and the counts of Bifidobacterium and Lactobacillus of the patients in compensated cirrhosis group were decreased(P<0.05).Compared with compensated cirrhosis group, the patients in decompensated cirrhosis group had higher Enterobacter and Enterococcus counts and lower Bifidobacterium and Lactobacillus counts(P<0.05).Compared with healthy control group, the patients in compensated cirrhosis group had higher levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),and total bilirubin(TBIL),and lower levels of albumin(ALB)(P<0.05).Compared with compensated cirrhosis group, the patients in decompensated cirrhosis group had higher ALT,AST,and TBIL levels and lower ALB levels(P<0.05).Compared with healthy control group, the patients in compensated cirrhosis group had lower WBC,PLT,and NE levels(P<0.05).Compared with compensated cirrhosis group, the patients in decompensated cirrhosis group had lower WBC,PLT,and NE levels(P<0.05).Compared with healthy control group, the patients in compensated cirrhosis group had higher levels of hyaluronic acid(HA) and type Ⅲ procollagen peptide(PⅢP)(P<0.05).Compared with compensated cirrhosis group, the patients in decompensated cirrhosis group had higher HA and PⅢP levels(P<0.05).Compared with healthy control group, the patients in compensated cirrhosis group had higher levels of interleukin-6(IL-6),interleukin-8(IL-8),tumor necrosis factor-α(TNF-α),and lipopolysaccharide(LPS)(P<0.05).Compared with compensated cirrhosis group, the patients in decompensated cirrhosis group had higher levels of IL-6,IL-8,TNF-α,LPS,Toll-like receptor 4(TLR4),nuclear factor-kappa B(NF-κB),c-Jun N-terminal kinase(JNK),and p38 mitogen-activated protein kinase(p38 MAPK)(P<0.05).Enterobacter, Enterococcus, Bifidobacterium, Lactobacillus, LPS,and TLR4 were risk factors for cirrhosis complicated by ascites.Conclusion The clinical characteristics of patients with cirrhosis are associated with intestinal flora.The indicators such as inflammation, immunity, and intestinal flora are closely related to the development of cirrhosis with ascites.Enterobacter, Enterococcus, Bifidobacterium, Lactobacillus, LPS,and TLR4 are the risk factors for cirrhosis complicated by ascites.