Translated Abstract
Objectives:
The objectives of this experiment are to study the prevention and treatment effect of Xiaochaihu decoction on Acute-on-chronic liver failure and the dose-effect relationship, to discuss the mechanism of this preventive effect and to provide theoretical and experimental basis for the clinical application of Xiaochaihu decoction in the treatment of Acute-on-chronic liver failure.
Methods:
1. 130 Wistar rats of healthy and clean grade were chosen and randomly divided into 7 groups A, B, C, D, E, F and G. Normal feeding without intraperitoneal injection was seen in the normal control group (Group A). Intraperitoneal injection of normal saline with the same dose was given to the rats in sham experimental group (Group B). Intraperitoneal injection of 40% mixed solution of carbon tetrachloride and vegetable oil was given to the rats in Acute-on-chronic liver failure model group (Group C), Xiaochaihu decoction groups (Group D, E, F) and polyene phosphatidyl choline group (Group G). once every three days and for 6 weeks altogether.
2. During the intraperitoneal injection, low, medium and large doses of Xiaochaihu decoction were lavaged, respectively, to rats in group D, E and F for intervention. Polyene phosphatidyl choline solution was lavaged to the rats in group G. Nothing was lavaged to the rats in group A and normal saline of the same dose was lavaged to group B and group C. The above intervention was carried out once every day and lasted till the end of the experiment.
3. Five rats in each group were randomly chosen three days after the last intraperitoneal injection. And then blood was drawn to detect ALT AST and TBIL. Livers were gathered to get HE-staining and Masson-staining,in order to know whehter the liver injury model is successful or not.
4. To build the animal model of liver failure, intraperitoneal injection of D-Gal (700mg/kg) and LPS (10 ug/kg), was given to the remaining groups of rats.
5. The serum of rats was collected at different periods of time to test their liver functions and TNF-α levels. Liver pathology specimens were drawn from the rats that had been put to death and the changes in liver tissues were observed under light microscope after HE-staining.
6. TUNEL test was conducted on the liver pathological specimens to obtain the data on hepatocyte apoptosis index and carry out a correlation analysis between apoptosis index (AI) and TNF-α.
Results:
1. The preventive effect of Xiaochaihu decoction on experimental liver injury and the dose-effect relationship
1) The liver pathological specimens in rats which get carbon tetrachloride intraperitoneal injection 6 weeks showed liver fibrosis, indicating the successful building of a chronic liver injury model.
2) In comparison to the model group, the percentage differences of liver fibrosis areas in the Xiaochaihu decoction treatment group were statistically significant( P ＜ 0.05).The pair-wise comparison between Xiaochaihu decoction treatment groups showed that the percentages of liver fibrosis areas were statistically different (P ＜ 0.05). Besides, the liver fibrosis area decreased with the increase of Xiaochaihu decoction doses, displaying the existence of certain dose-effect relationship.
3) In contrast with the model group, ALT, AST and TBIL in the Xiaochaihu decoction treatment group and the polyene phosphatidyl choline treatment group were statistically different (P ＜ 0.05).As indicated by the pair-wise comparison between Xiaochaihu decoction treatment groups, the ALT and AST levels were statistically different(P ＜ 0.05). Values of the above observational indexes decline gradually with the increase of Xiaochaihu decoction doses, illustrating the existence of dose-effect relationship. In comparison to group E and F, the TBIL level in group D was statistically different(P ＜ 0.05). But the TBIL level difference between group E and group F was not statistically significant(P = 0.161).
2. The preventive effect of Xiaochaihu decoction on Acute-on-chronic liver failure and the dose-effect relationship
1) After the intraperitoneal injection of D-Gal and LPS on the rats which suffered from experimental liver injuries, the liver pathology specimens of the rats displayed different degrees of liver apoptosis. It indicated the successful building of the Acute-on-chronic liver failure model.
2) The comparison between treatment groups and model groups showed that the coefficients of liver apoptosis were statistically different(P ＜ 0.05). And the coefficients exhibited a gradual decline with the increase of Xiaochaihu decoction doses, indicating the existence of a certain dose-effect relationship.
3) In contrast with model groups, ALT, AST, TBIL and TNF-α in treatment groups were statistically different(P ＜ 0.05). The above indexes declined gradually as the dose of Xiaochaihu decoction dose increased, implying the existence of a dose-effect relationship.
4) The TNF-α level of rats in each group showed a liner correlation with the apoptosis index of liver(r = 0.63, P ＜ 0.0001).
Conclusions:
1. The Xiaochaihu decoction exerts a protective effect on the liver cells of rats with experimental liver injuries induced by carbon tetrachloride and mitigates liver fibrosis to a minor degree. Besides, certain dose-effect relationship exists between this protective effect and Xiaochaihu decoction doses. The best therapeutic effect can be achieved with the Xiaochaihu decoction dose recorded in the classical prescription.
2. The Xiaochaihu decoction can significantly reduce the liver injury degree of Acute-on-chronic liver failure and alleviate liver apoptosis. This effect shows a correlation with the doses of Xiaochaihu decoction. The best therapeutic effect can be achieved with the Xiaochaihu decoction dose recorded in the classical prescription.
3. Decrease the release of TNF-α by inhibiting the activation of mononuclear macrophages, thereby reducing liver apoptosis. This is the possible mechanism through which the Xiaochaihu decoction exerts a protective effect on Acute-on-chronic liver failure.
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