“香胃联合方组”对肝硬化大鼠血浆内毒素水平的影响

Effect of Xiangwei comined decoction on plasma endotoxin level in cirrhotic rats

目的观察“香胃联合方组”对肝硬化大鼠血浆内毒素水平的影响,探讨“香胃联合方组”降低血浆内毒素的作用机制。方法 85只Wistar大鼠随机抽取10只为正常组,10只为香胃联合预防组(预防组),10只为乳果糖预防组(预对组),55只为纯造模组。采用四氯化碳复合因素制造肝硬化模型。造模期间,预防组给予香砂六君子汤6.875 g·kg^-1·d^-1、胃苓汤7.188 g·kg^-1·d^-1隔日交替灌胃;预对组给予乳果糖口服溶液2.083 g·kg^-1·d^-1灌胃,每日一次;正常组和纯造模组每日一次给予等量生理盐水1 ml/100 g灌胃。造模持续8周。造模过程中大鼠死亡19只。造模成功后,正常组8只,预防组9只,预对组8只,纯造模组随机分为模型组9只,香胃联合方组(香胃组)8只,香砂六君子汤组(香砂组)8只,胃苓汤组(胃苓组)8只,乳果糖对照组(治对组)8只。预防组、预对组继续给予相应的药灌胃;正常组和模型组继续给予等量生理盐水灌胃;香胃组给予香砂六君子汤6.875 g·kg^-1·d^-1、胃苓汤7.188 g·kg^-1·d^-1隔日交替灌胃;香砂组给予香砂六君子汤6.875 g·kg^-1·d^-1灌胃,每日一次;胃苓组给予胃苓汤7.188 g·kg^-1·d^-1灌胃,每日一次;治对组给予乳果糖口服溶液2.083 g·kg^-1·d^-1灌胃,日一次;12周后观察各组血浆内毒素、血清肠脂肪酸结合蛋白(IFABP)、D-乳酸及小肠组织分泌型免疫球蛋白A(sIgA)、Occludin mRNA水平;苏木精-伊红(HE)染色观察小肠组织病理表现。结果血浆内毒素、血清D-乳酸、IFABP水平,正常组<预防组<预对组<香胃组<香砂组、胃苓组<治对组<模型组,除胃苓组和香砂组之间比较无统计学差异外(P>0.05),其余各组间两两比较差异均有统计学意义(P<0.05,P<0.01)。小肠组织sIgA、Occludin mRNA水平,正常组>预防组>预对组>香胃组>胃苓组>香砂组>治对组>模型组,除胃苓组和香砂组之间比较无统计学差异外(P>0.05),其余各组间两两比较差异均有统计学意义(P<0.05,P<0.01)。结论香胃联合方组通过减少肝硬化大鼠血清D-乳酸、小肠IFABP水平,降低肠黏膜通透性,增加紧密连接Occludin蛋白表达,提高sIgA的分泌水平,从而降低肝硬化大鼠血浆内毒素水平。

Objective To observe the influence of Xiangwei combined decoction on plasma endotoxin level in cirrhotic rats, and to investigate the mechanism of Xiangwei combined decoction in reducing plasma endotoxin. Methods Eightyfive Wistar rats were randomly selected, 10 as normal group, 10 as Xiangwei combined prevention group ( prevention group), 10 as lactulose prevention group ( precontrol group),and 55 as pure model group. The model of liver cirrhosis was established by carbon tetrachloride combined factors. During the modeling period, the preventive group was given 6. 875 g·kg^-1·d^-l Xiangsha Liujunzi decoction and 7. 188 g·kg^-1·d^-l Weiling decoction alternately every other day;the pretreated group was given 2. 083 g·kg^-1·d^-l lactulose oral solution once a day;the normal group and the pure model group were given 1 ml/100 g normal saline once a day. The model lasted for 8 weeks. Nineteen rats died during the process of establishing the model. After successful modeling,there were 8 rats in the normal group,9 in the prevention group and 8 in the pre-pair group,and the 41 rats in the pure model group were randomly divided into the model group (n =9),Xiangwei combined prescription group ( Xiangwei group,n =8),Xiangsha Liujunzi decoction group ( Xiangsha group,n =8), Weiling decoction group ( Weiling group, n = 8), and the lactulose control group (treatment-control group, n = 8). The prevention group and the pre-control group continued to be given corresponding medicines;the normal group and the model group continued to be given the same amount of saline;the Xiangwei group was given Xiangsha Liujunzi decoction 6. 875 g·kg^-1·d^-l , Weiling decoction 7. 188 g·kg^-1·d^-l alternately every other day;the Xiangsha group was given Xiangsha Liujunzi decoction 6. 875 g·kg^-1·d^-l ,once a day;the Weiling group was given Weiling decoction 7. 188 g·kg^-1·d^-l,once a day. The treatment-control group was given lactulose oral solution 2. 083 g·kg^-1·d^-l by gastric perfusion once a day. After 12 weeks, the levels of plasma endotoxin, serum intestinal fatty acid binding protein (IFABP), D-lactic acid and secretory immunoglobulin A ( slgA) and Occludin mRNA in small intestine tissues were observed, and the pathological features of small intestine tissues were observed by hematoxylin-eosin ( HE) staining. Result The levels of plasma endotoxin, serum D-lactic acid and IFABP in normal group, prevention group, pre-control group, Xiangwei group, Xiangsha group/Weiling group, treatment-control group and model group increased in order, and there were significant differences between the other two groups, except that there was no significant difference between Weiling group and Xiangsha group (P >0. 05 ). The levels of slgA and Occludin in small intestine tissues were decreased in the order of normal group, prevention group, pre-control group, Xiangwei group, Weiling group, Xiangsha group, treatment-control group and model group, and there were significant differences between the other two groups, except that there was no significant difference between Weiling group and Xiangsha group (P >0. 05). Conclusion Xiangwei combined decoction can reduce the level of serum D-lactic acid and small intestinal IFABP,reduce intestinal mucosal permeability,increase the expression of tight junction Occludin protein, and increase the secretion level of slgA in cirrhotic rats, thereby reducing the level of plasma endotoxin in cirrhotic rats.