| BackgroundIntrahepatic cholestasis is an important risk factor of the speeding up of cirrhosis of the liver. There are significant changes in serum biochemical parameters and in organizational structure of liver. It was found by molecular biology technology that the bile salt export pump(BSEP) in liver cells and bile duct cell membrane, and the membrane transporter NTCP, are important molecular pathways for regulation of bile secretion and excretion. The abnormal regulation of BSEP and NTCP led to the intrahepatic cholestasis directly. Cholestatic jaundice are equal to the jaundice in Traditional Chinese Medicine (TCM). Treatments in TCM have showed unique effect and advantages. In clinical work normalizing the gallbladder function to treat jaundice is an effective way. But the mechanism of normalizing the gallbladder function was not illuminated in reported researches. Most of these researches were simple clinical observation. The number of experimental reports and the reported indicators were relatively little. Most of the clinical observations showed a lack of rigorous design, and of control group. The data and statistics method were relatively poor. And most of them were non-multi-center studies. The criteria of diagnosis and efficacy was not standardized.PurposeTo make the intrahepatic cholestasis rat model with dampness-heat syndrome by the composite factors as hot and damp environment and fat dietary and etiopathogenisis. By the combination of disease and syndrome animal model, the effect of herbal compound, a single herb and the active ingredients of herb could be studied by the expressions of BSEP(bile salt export pump) and NTCP. The serum biochemical parameters and structural changes in the liver were observed at the same time. This could provide a theoretical basis for the treatments of jaundice by normalizing the gallbladder function, and so to discuss the targets.Subjects and MethodsExperiment on rats:1 modeling:kept the rats in hot and humid boxes in the long summer festival, and fed them with the mixture of glucose and pig fat to form damp-heat syndrome. Then usedα-naphthyl ester isothiocyanate (ANIT) to form the intrahepatic cholestasis rat model.2 grouping and intervention:150 three-month-old male SD rats were randomly divided into 5 groups, each group had 15 rats. Here were the five groups:normal control group, model group, YCHT group, single-flavor capillaris group, and matrine groupCompound YCHT Group:Lavaged the rats with 100% Yin Chen Hao Decotion 2mL /100g everyday from the 8th afternoon of modeling to the 28th days after modeling.Single herb Artemisia Group:Lavaged the rats with 100% Artemisia 2mL/ 100g everyday from the 8th afternoon of modeling to the 28th days after modeling.Matrine:Lavaged the rats with 100% Matrine 2mL/100g everyday from the 8th afternoon of modeling to the 28th days after modeling.Normal control group and model group were lavaged with the 0.9% saline 2mL/100g.3 indicatrix and methods:3.1 Analyzed the ALT, AST, ALP, TB, DB in rats serum by automatic biochemical analyzer.3.2 Got the liver tissue to do routine pathological examination in HE staining, to observe the structural changes in liver tissue by microscopy.3.3 detected the NTCP and BSEP in the liver tissue by the immunohistochemical staining.3.4 detected the BSEP and NTCP in liver tissue by fluorescent real-time quantitative reverse transcription polymerase chain reaction (Real-time RT-PCR). Clinical research:Clinical research on efficacy of normalizing the gallbladder function to treat jaundice:Collected 60 patients who met relevant standards from Digestion Department in First Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine, Guangzhou Eighth People's Hospital. This research used a randomized, positive control method. All patients were divided into two groups. The control group were treated with conventional symptomatic treatment, intravenous think Mattel and glutathione 1.0g 1.2g; and took ursodeoxycholic acid 250mg/time,2 times/d orally. The herb group took Yin Chen Hao Decotion Jia Wei. (The basic formula is capillaris, gardenia, rhubarb, red peony root, turmeric, money grass, Polygonum etc) orallyChanges in clinical symptoms and signs, tongue, pulse, liver function (TB, DB, ALT, GGT), clinical comprehensive efficacy, and TCM syndromes were observed. Evaluated the clinical efficacy of normalizing the gallbladder function to treat jaundice on intrahepatic cholestasis patients and provided clinical data.Result1 After modeling, the symptoms and signs of cholestasis and damp-heat syndrome, the bilirubin, total bile acid, serum liver enzymes and the pathological changes in the tissue could be improved in the herb groups.2 Research on the changes of BSEP and NTCP in normalizing the gallbladder function to treat jaundice showed that:expression of BSEP and NTCP in model group decreased significantly,while the ones'expression in herb groups increased but still not recover to the nomal level. There were significant differences between the herb groups and the model group by statistics.3 The result of clinical research showed that the improvements of clinical efficacy in the herb group were similar to the control group. The serum bilirubin and liver enzymes in both groups improved and there was no difference between them. None of them showed side effects In the improvements of syndromes'score and syndromes'efficacy, the herb group showed better efficacy.Conclusion1 According to the causes of disease, symptoms, signs, laboratory indicators and treatment efficacy, the rat model in this reseach fit both syndrome in Chinese Medicine and intrahepatic cholestasis. It was more satisfying integrative medicine model of disease and syndrome. It was confirmed that intervention by normalizing the gallbladder function to treat jaundice could effectively improve symptoms, signs and bilirubin, total bile acid and liver function of the rats model, and reduce the pathological damage of liver tissue.2 In this research, the regulation of NTCP and BSEP's expression in rats' liver tissue was explored by immunohistochemistry, RT-PCR analysis and other techniques. It was found that expression of BSEP and NTCP was significantly reduced in intrahepatic cholestasis rat model. The intervention of normalizing the gallbladder function to treat jaundice in Chinese Medicine could help increase the expression of BSEP and NTCP, increase bile salt uptake and excretion in liver cell, and in particular, promote functional recovery of bile salt transport system. The result provided the clinical theory basis for treatment of normalizing the gallbladder function to treat jaundice in intrahepatic cholestasis patients.3 The representative decotion of normalizing the gallbladder function to treat jaundice was Jia Wei Yin Chen Hao Decotion. The clinical efficacy of Jia Wei Tin Chen Hao Decotion to intrahepatic cholestasis patients was satisfactory in this research. Compared with western medicine control group, the scores of syndrome and efficacy were improved better in herb group. This provided reliable evidence for application of normalizing the gallbladder function to treat jaundice...
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