Qinggan Huoxue Recipe suppresses epithelial-to-mesenchymal transition in alcoholic liver fibrosis through TGF-ß1/Smad signaling pathway.

AIM: To investigate the mechanism by which Qinggan Huoxue Recipe (QGHXR) inhibits epithelial-to-mesenchymal transition (EMT) in rats with alcoholic liver fibrosis (ALF). METHODS: A total of 75 male SD rats were used to induce ALF. Serum biochemical indicators, including alanine aminotransferase, aspartate aminotransferase, laminin and hyaluronidase, were measured. Liver histopathological changes were evaluated using hematoxylin-eosin and Sirius red staining. EMT was examined by analyzing the expression of the epithelial marker E-cadherin and the mesenchymal markers vimentin and fibronectin using RT-PCR and Western blot. The inhibitory effect of QGHXR on EMT markers, as well as its effect on molecules associated with the transforming growth factor (TGF)-ß1/Smad signaling pathway, including TGF-ß1, Smad3, snail, occludin, ZO-1 and claudin, was also examined. RESULTS: Compared with normal control rats, ALF rats exhibited a decrease in E-cadherin levels (mRNA: ALF 0.16 ± 0.05 vs control 1.00 ± 0.08; protein: ALF 0.09 ± 0.05 vs control 0.70 ± 0.17, P < 0.01) and an increase in vimentin and fibronectin levels (mRNA: 11.43 ± 0.39 vs 1.00 ± 0.19 and 9.91 ± 0.34 vs 1.00 ± 0.44, respectively, P < 0.01; protein: 1.13 ± 0.42 vs 0.09 ± 0.03 and 1.16 ± 0.43 vs 0.09 ± 0.00, respectively, P < 0.01). This indicates that EMT occurred in ALF rats. In addition, the TGF-ß1/Smad signaling pathway was activated in ALF rats, as evidenced by the increase in TGF-ß1 and snail levels (mRNA: 1.76 ± 0.12 vs 1.00 ± 0.05 and 6.98 ± 0.41 vs 1.00 ± 0.10, respectively, P < 0.01; protein: 1.43 ± 0.05 vs 0.12 ± 0.03 and 1.07 ± 0.29 vs 0.07 ± 0.02, respectively, P < 0.01) and the decrease in Smad3 levels (mRNA: 0.05 ± 0.01 vs 1.00 ± 0.12, P < 0.01; protein: 0.06 ± 0.05 vs 0.89 ± 0.12, P < 0.01). Furthermore, levels of the tight junction markers occludin, ZO-1 and claudin decreased in ALF rats compared with healthy control rats (mRNA: 0.60 ± 0.09 vs 1.00 ± 0.12, 0.11 ± 0.00 vs 1.00 ± 0.12 and 0.60 ± 0.01 vs 1.00 ± 0.08, respectively, P < 0.01; protein: 0.05 ± 0.01 vs 0.87 ± 0.40, 0.09 ± 0.05 vs 0.89 ± 0.18 and 0.04 ± 0.03 vs 0.95 ± 0.21, respectively, P < 0.01). In ALF rats treated with QGHXR, E-cadherin levels increased (mRNA: QGHXR 0.67 ± 0.04 vs ALF model 0.16 ± 0.05, P < 0.01; protein: QGHXR 0.66 ± 0.21 vs ALF model 0.09 ± 0.05, P < 0.01), and vimentin and fibronectin levels decreased (mRNA: 6.57 ± 1.05 vs 11.43 ± 0.39 and 1.45 ± 1.51 vs 9.91 ± 0.34, respectively, P < 0.01; protein: 0.09 ± 0.03 vs 1.13 ± 0.42 and 0.10 ± 0.01 vs 1.16 ± 0.43, respectively, P < 0.01). In addition, QGHXR inhibited the expression of TGF-ß1 and increased the expression of Smad3 (mRNA: 1.03 ± 0.11 vs 1.76 ± 0.12, 0.70 ± 0.10 vs 0.05 ± 0.01, respectively, P < 0.05 and P < 0.01; protein: 0.12 ± 0.03 vs 1.43 ± 0.05 and 0.88 ± 0.20 vs 0.06 ± 0.05, respectively, P < 0.01). QGHXR treatment also reduced the levels of the EMT-inducing transcription factor snail (mRNA: 2.28 ± 0.33 vs 6.98 ± 0.41, P < 0.01; protein: 0.08 ± 0.02 vs 1.07 ± 0.29, P < 0.01) and increased the occludin, ZO-1 and claudin levels (mRNA: 0.73 ± 0.05 vs 0.60 ± 0.09, 0.57 ± 0.04 vs 0.11 ± 0.00 and 0.68 ± 0.03 vs 0.60 ± 0.01, respectively, P < 0.01, P < 0.01 and P < 0.05; protein: 0.92 ± 0.50 vs 0.05 ± 0.01, 0.94 ± 0.22 vs 0.09 ± 0.05 and 0.94 ± 0.29 vs 0.04 ± 0.03, respectively, P < 0.01). The effects of QGR and HXR on the TGF-ß1/Smad signaling pathway were similar to that of QGHXR; however, the QGR- and HXR-induced changes in vimentin mRNA levels, the QGR-induced changes in fibronectin mRNA levels and the HXR-induced changes in snail and TGF-ß1 mRNA levels were not significant. CONCLUSION: Qinggan Huoxue Recipe inhibits EMT in ALF rats by modulating the TGF-ß1/Smad signaling pathway, suggesting that the mechanism underlying the amelioration of ALF induced by QGHXR is associated with this pathway.

Chinese medicinal formula, Qinggan Huoxue Recipe protects rats from alcoholic liver disease via the lipopolysaccharide-Kupffer cell signal conduction pathway.

The Chinese medicinal formula, Qinggan (QG) Huoxue (HX) Recipe (R) exerts a range of pharmacological effects, including reversible steatosis, decreased levels of inflammatory cytokines and lipid peroxidation resistance. The aim of the present study was to determine the specific mechanisms of QGHXR hepatoprotection through the lipopolysaccharide-Kupffer cell (LPS-KC) signal conduction pathway in rats with alcoholic liver disease (ALD). ALD rats were exposed to the compound factors, QGR and HXR. Hematoxylin and eosin staining was conducted to evaluate the pathological changes in the liver following QGHXR treatment and an enzyme-linked immunosorbent assay was performed to measure the content of tumor necrosis factor (TNF)-α in the plasma. Immunohistochemical staining was conducted to examine the expression of cell differentiation antigen (CD) 68 and 14. In addition, western blot analysis and reverse transcription-polymerase chain reaction were used to measure the expression of Toll-like receptor 4 (TLR4), phosphorylated-extracellular regulated protein kinases (p-ERK), nuclear factor (NF)-κB, CD14 and TNF-α. Following stimulation with the compound factors, the rats exhibited increased alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels, as well as marked pathological changes. Furthermore, the related molecules in the LPS-KC pathway were upregulated and QGHXR was identified to be effective in the LPS-KC signal conduction pathway in the ALD rats. QGHXR was superior to QGR and HXR in reducing the serum ALT and AST levels, regulating CD14, TLR4, NF-κB, ERK and TNF-α as well as improving the pathological changes. The results indicated that QGHXR therapy may provide a novel strategy for treating ALD via regulation of the related molecules in the LPS-KC signaling pathway.

Chinese medicine formula lingguizhugan decoction improves Beta-oxidation and metabolism of Fatty Acid in high-fat-diet-induced rat model of Fatty liver disease.

Lingguizhugan decoction (LGZG), a classic traditional Chinese medicine (TCM) formula, has been used to treat obesity and hyperlipidemia in recent years, but the related mechanisms underlying the regulation of lipid metabolism by LGZG are not clear yet. Here, we reported the effectiveness and possible mechanisms of LGZG on rats with fatty liver disease induced by high-fat diet (HFD). Our results demonstrated that LGZG significantly attenuated HFD-induced fatty liver disease, as measured by body weight, liver index, epididymal fat pad-body weight ratio (EFP/BW), liver injury, and hepatic triglycerides (TG) probably through increasing serum thyroid hormone levels, improving beta-oxidation (via modulation of TR ß 1 and CPT1A expression), metabolism and transport (through modulation of SREBP-1c, ACSL and ApoB100 expression) of fatty acid. In addition, we discovered the herbal combination with the properties of warming yang to relieve water retention in the formula and proposed the biological basis of LGZG conventional effect via further study on disassembled formula. This study, for the first time, revealed the mechanisms through which LGZG regulates lipid metabolism. Furthermore, our study suggested that it might be feasible to understand the scientific implications of TCM from the perspective of classic formulas' conventional efficacy.

Berberine reducing insulin resistance by up-regulating IRS-2 mRNA expression in nonalcoholic fatty liver disease (NAFLD) rat liver.

This study was performed to investigate the molecular mechanism and the therapeutic effect of berberine on nonalcoholic fatty liver disease (NAFLD). Rat models were given a high-fat diet (42% kcal) until they developed NAFLD, then were given normal saline (n=10), berberine (n-=10) at 187.5mg/kg/day, or pioglitazone (n=10) at 10.0mg/kg/day intragastrically for 4 weeks, respectively, and evaluated by hyperinsulinemic euglycemic clamping for insulin sensitivity. Serum biochemical markers and liver triglyceride (TG) were analyzed, real-time RT-PCR for mRNA expression and western blotting for protein expression of insulin receptor (IR) and insulin receptor substrate-2 (IRS-2) in liver tissues were performed, and hepatic histopathology in the rat models with NAFLD at the end of treatment was compared with normal controls (n=10). The NAFLD rats developed insulin resistance, showing increased fasting blood glucose and insulin levels, decreased glucose infusion rate, increased weight of epididymal fat (g/100g body weight), obvious hepatic steatosis and inflammation, and down-regulated IRS-2 mRNA and protein levels compared with normal controls (all P<0.05). In comparison with those treated with saline, model rats treated with berberine or pioglitazone underwent significant recovery, including up-regulated IRS-2 mRNA and protein (all P<0.05). Our results indicate that berberine may improve insulin resistance of NAFLD by up-regulating mRNA and protein levels of IRS-2, a key molecule in the insulin signaling pathway, suggesting that berberine may be used to treat NAFLD.

[Regulatory effects of Qinggan Huoxue Recipe on matrix metalloproteinases of alcoholic liver fibrosis rats].

OBJECTIVE: To study the action mechanism of Qinggan Huoxue Recipe (QGHXR) and its disassembled recipes for treatment of alcoholic liver fibrosis (ALF) by observing their regulation on the expressions of matrix metalloproteinases (MMPs) and type 1 tissue inhibitor of metalloproteinase (TIMP-1). METHODS: 130 male SD rats were randomly divided into the blank control group (n=10), the CCI4 group (n=10), and the modeling group (n=110). Rats in the modeling group were intervened by complex factors dominated as alcohol. Eight weeks later they were randomly divided into 4 subgroups, i.e., the model group (n=25), the QGHXR group (n= 15), the Qinggan Recipe (QGR) group (n=15), and the Huoxue Recipe (HXR) group (n=15). Eight model rats were selected for pathological analysis to monitor the development of the modeling at the 4th, 8th, and 10th week of the experiment. The rest rats died during the modeling. Corresponding medicines were given to these treatment groups (at the dose of 4.75 g/kg, 1.50 g/kg, and 3.25 g/kg). All rats were killed at the end of the 12th week. The protein and mRNA expressions of MMP-2, MMP-9, and TIMP-1 were detected using Western blotting, fluorescent quantitative polymerase chain reaction, and immunofluorescence method. RESULTS: Compared with the blank control group, the expressions of MMP-2, MMP-9, and TIMP-1 significantly increased in the model group (1.81 +/- 0.28 versus 0.53 +/- 0.04, 1.60 +/- 0.16 versus 0.45 +/- 0.05, 1.20 +/- 0.02 versus 0.35 +/- 0.07, P < 0.01). Compared with the model group, QGHXR and its disassembled recipes all could decrease the protein and mRNA expressions of TIMP-1 (0. 56 +/- 0.05, 0.67 +/- 0.02, 0.70 +/- 0.02 versus 1.20 +/- 0.02, P < 0.05), increase the expressions of MMP-2 and MMP-9 (4.18 +/- 0.53, 2.70 +/- 0.40, 2.38 +/- 0.22 versus 1.81 +/- 0.28, 3.31 +/- 0.06, 2.56 +/- 0.20, 1.87 +/- 0.05 versus 1.60 +/- 0.16, P < 0.05, P < 0.01). QGHXR was superior to its disassembled recipes (P < 0.05, P < 0.01). CONCLUSION: The action mechanisms of QGHXR and its disassembled recipes might possibly be correlated with regulating MMPs.

[Relationship between the phenotypes and functions of peripheral blood dendritic cells and the different spleen deficiency syndrome types in patients with chronic hepatitis B].

OBJECTIVE: To study the phenotypes and functions of dendritic cells (DCs) derived from peripheral blood monocytes of chronic hepatitis B (CHB) patients with different traditional Chinese medicine (TCM) syndrome types, and to explore the relationship between TCM syndrome type and DC functions. METHODS: Sixty CHB patients were included in this study. All the CHB patients were divided into spleen deficiency and liver stagnation, spleen deficiency and dampness-heat and deficiency of both spleen and kidney groups according to TCM syndrome diagnosis standard. There were 20 cases in each group, and ten healthy people were included as normal control. The volunteer's peripheral blood was collected for monocyte separation, biochemical test and hepatitis B virus DNA loads detection. DCs were induced and isolated from peripheral blood monocytes, and then the expressions of surface markers CD80, CD86, CD1a and HLA-DR were detected by flow cytometric analysis method. Interleukin-10 (IL-10) production of the DCs was quantified by enzyme-linked immunosorbent assay. RESULTS: The proliferation of DCs in the CHB patients was slower than that in the healthy volunteers (P<0.05). The expressions of DC surface molecules such as CD80, CD86, and CD1a were obviously decreased in the CHB patients as compared with those in the healthy volunteers (P<0.05). More over, expressions of DC surface molecules were different among CHB patients with different TCM syndrome types. The positive expressions of CD80, CD1a, and HLA-DR in the CHB patients with spleen deficiency and liver stagnation were obviously higher than those in the CHB patients with deficiency of both spleen and kidney (P<0.05), and the CD1a expression in the CHB patients with spleen deficiency and dampness-heat was higher than that in the CHB patients with deficiency of both spleen and kidney (P<0.05). In DC culture supernatant, the IL-10 concentration of the CHB patients with deficiency of both spleen and kidney was higher than that of the CHB patients with spleen deficiency and liver stagnation (P<0.05), and the IL-10 concentrations of the CHB patients with different TCM syndrome types were higher than that of the healthy volunteers (P<0.05). CONCLUSION: During the pathogenic course of CHB, the phenotypes and functions of DCs are different in CHB patients with different TCM syndrome types. It suggests that there is a correlation between TCM syndrome type and body immunity function.

[Effects of Qinggan Huoxue Recipe and its separated recipes on urokinase-type plasminogen activator and plasminogen activator inhibitor-1 fibrinolytic system in rats with alcoholic liver fibrosis].

OBJECTIVE: To study the action mechanisms of Qinggan Huoxue Recipe (QGHXR), a compound traditional Chinese herbal medicine, and its separated recipes by observing their effects on expressions of urokinase-type plasminogen activator (uPA) and plasminogen activator inhibitor-1 (PAI-1) in rats with alcoholic liver fibrosis (ALF). METHODS: A total of 150 SD rats were divided into three groups: blank group, carbon tetrachloride (C(4)) group and ALF-inducing group. Rats in the ALF-inducing group were administered with a mixture diet (56% alcohol 10 mL/kg, corn oil 2 mL/kg, pyrazole 25 mg/kg) once daily and intraperitoneally injected with 0.3 mL/kg 25% solution of C(4) in olive oil twice weekly. The C(4) group was intraperitoneally injected with equal volume of C(4) and olive oil as the ALF-inducing group and ingested normal saline (12 mL/kg per day). The blank group was intraperitoneally injected with and ingested saline in equal volumes of the above. At the end of the eighth week, the survived rats in the ALF-inducing group were divided into four subgroups: untreated group, QGHXR group, Qinggan Recipe (QGR) group and Huoxue Recipe (HXR) group. The three treated groups were given corresponding drugs respectively (4.75, 1.5, 3.25 g/kg). The blank group, CCl(4) group and untreated group were given normal saline in equal volume (5 mL/kg per day). All rats were anaesthetized and killed at the end of the twelfth week. The activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in the serum were analyzed. Pathological changes in liver tissues were observed by hematoxylin and eosin staining and Masson staining. The expressions of uPA and PAI-1 were evaluated with Western blotting, immunofluorescence, and real-time polymerase chain reaction. RESULTS: There existed obvious liver fibrosis in liver tissues in the untreated group as compared with the blank group (P<0.01), and the activities of ALT and AST and the expressions of uPA and PAI-1 also increased in the untreated group. QGHXR and its separated recipes could improve the degree of liver fibrosis (P<0.01). QGHXR and its separated recipes could degrade the activity of ALT as compared with the untreated group; QGHXR and its separated recipes advanced the expression of uPA, and decreased the expression of PAI-1 significantly as compared with the untreated group. The effect of QGHXR was the best among the three recipes. CONCLUSION: QGHXR and its separated recipes may improve ALF in rats by decreasing the expression of PAI-1 and advance the expression of uPA. The effect of QGHXR is the best among them.

[Distribution pattern of traditional Chinese medicine syndromes in 793 patients with fatty liver disease].

OBJECTIVE: To study the distribution pattern of traditional Chinese medicine (TCM) syndromes in fatty liver disease. METHODS: A multicenter and large sample survey was carried out by adopting the model of "combining disease with syndrome". A TCM syndrome information database was established by EPidata 3.1 software. The distribution pattern of TCM syndromes in fatty liver was studied by factor analysis and cluster analysis methods with SPSS 13.0 software. RESULTS: The basic syndromes of fatty liver included insufficiency of liver and kidney, flaring fire due to yin deficiency, liver-qi stagnation and spleen deficiency, spleen deficiency, spleen deficiency and dampness stagnation, mild syndrome of internal accumulation of damp-heat, blood stasis, severe syndrome of internal accumulation of damp-heat, and internal stagnation of phlegm-dampness. Single syndrome and combination of two to four basic syndromes were common in fatty liver disease. The syndrome of spleen deficiency and dampness stagnation was the most frequent one when its pathogenesis was simple, while the syndrome of insufficiency of liver and kidney was most frequent one when the pathogenesis was complicated. A total of 108 patients (13.6%) had no obvious symptoms, 46 patients (5.8%) were classified into the pattern of non-categorization, and the other patients were classified into five syndromes including phlegm accumulating with stagnation due to spleen deficiency (11.5%, 91/793), yin deficiency of liver and kidney (18.5%, 147/793), retention of phlegmatic dampness due to spleen deficiency (32.0%, 254/793), internal accumulation of damp-heat due to spleen deficiency (10.2%, 81/793), and damp obstruction due to liver-qi stagnation and spleen deficiency (8.3%, 66/793). CONCLUSION: Multi-element analysis reveals the distribution pattern of TCM syndromes in fatty liver disease, which is worthy of further study. The basic pathogenesis is spleen deficiency, and has a close correlation with the liver and kidney. The main pathogenesis factors are phlegm, dampness, blood stasis, heat and liver-qi stagnation. Yin deficiency of liver and kidney is a typical syndrome in fatty liver disease.

[Effects of Qinggan Huoxue Recipe and its separated recipes on the expression of tumor necrosis factor-alpha in rats with alcoholic liver injury].

OBJECTIVE: To study the effects of Qinggan Huoxue Recipe (QGHXR), the compound traditional Chinese herbal medicine, and its separated recipes on the expression of tumor necrosis factor-alpha (TNF-alpha) mRNA and serum TNF-alpha content in rats with alcoholic liver injury (ALI). METHODS: One hundred male Wistar rats were randomly divided into normal control group (n=10), carbon tetrachloride (CCl4) group (n=10) and ALI group (n=80). Rats in the ALI group were intragastrically administered mixed liquor twice a day and intraperitoneally injected with CCl4 twice a week for 6 weeks, rats in the normal control group were intragastrically administered normal saline, and rats in the CCl4 group were intraperitoneally injected with CCl4 and olive oil twice a week continuously. Two rats in the ALI group were sacrificed for histological observation per week. After 4-week modeling, the rats in the ALI group were randomly divided into QGHXR group, Qinggan Recipe (QGR) group, Huoxue Recipe (HXR) group (15 rats in each group), and the others belonged to the untreated group. After 2-week suitable drugs treatment, the activities of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were analyzed. Pathological changes in liver tissues were observed by HE staining. The content of plasma TNF-alpha was assayed by enzyme linked immunosorbent assay, and expression of TNF-alpha mRNA in the liver tissue was detected by reverse transcription polymerase chain reaction (RT-PCR). RESULTS: QGHXR and its separated recipes improved liver steatosis and inflammation, and in this regard, the QGHXR was superior to the QGR. QGHXR decreased the activity of serum ALT in rats with ALI, but QGR and HXR did not show significant effect in that. The three recipes decreased the activity of AST as compared with the untreated group, but there were no significant differences among the three treated groups. HXR and QGHXR down-regulated the expression of TNF-alpha mRNA in the liver tissue, but QGR did not show significant effect. HXR and QGHXR also decreased the content of plasma TNF-alpha, but QGR did not show significant effect in that. CONCLUSION: QGHXR and HXR may provide protection against ALI in rats through decreasing the production of TNF-alpha.

[Effects of puerarin on expressions of leptin receptor mRNA, phosphorylated Janus kinase 2/phosphorylated signal transducers and activators of transcription 3 proteins in the liver of rats with non-alcoholic fatty liver].

OBJECTIVE: To observe the effects of puerarin on the expressions of leptin receptor mRNA and phosphorylated Janus kinase 2 / phosphorylated signal transducers and activators of transcription 3 (P-JAK2/P-STAT3) proteins in the liver of rats with non-alcoholic fatty liver (NAFLD). METHODS: A rat model of NAFLD was successfully established by feeding high-fat diet. All SD rats were randomly divided into blank control group, untreated group, simvastatin-treated group and puerarin-treated group. After four-week treatment, the levels of hepatic triglyceride and total cholesterol were analyzed by using an automatic biochemical analyzer. The pathology of the liver tissue was observed by light microscopy. Serum leptin level was detected by enzyme-linked immunosorbent assay, and the expressions of leptin receptor mRNA and P-JAK2/P-STAT3 proteins in the liver of NAFLD rats were quantified by reverse transcription polymerase chain reaction (RT-PCR) and Western blot analysis respectively. RESULTS: Puerarin significantly decreased the levels of hepatic triglyceride and total cholesterol in NAFLD rats. Fat degeneration and inflammatory reaction in liver tissues of NAFLD rats were ameliorated after puerarin treatment. The serum leptin level was increased and the expressions of leptin receptor mRNA and P-JAK2/P-STAT3 proteins were up-regulated in puerarin-treated group. CONCLUSION: Puerarin can effectively attenuate liver lipid disorder and inflammation by improving the leptin resistance and enhancing the expressions of leptin receptor mRNA and P-JAK2/P-STAT3 proteins.

Effectiveness of Danning Tablet in patients with non-alcoholic fatty liver of damp-heat syndrome type: a multicenter randomized controlled trial.

OBJECTIVE: To evaluate the efficacy and safety of Danning Tablet (DNT) in patients with non-alcoholic fatty liver disease (NAFLD) of damp-heat syndrome type. METHODS: A multicenter, randomized, double-blinded and positive drug parallel controlled trial was performed. One hundred and thirty-five patients were enrolled into the study and divided into two groups: DNT-treated group (n=102) and ursodeoxycholic acid (UDCA)-treated group (n=33). Body mass index (BMI), principal symptoms, liver function, blood lipids, iconographic, and compositional parameters were measured before and after treatment, respectively. RESULTS: In the two groups, BMI, distress in hepatic region, fatigue, anorexia, liver function, blood lipids and iconographic parameters were significantly improved, and the improvements of BMI, distress in hepatic region were better in DNT-treated group than in UDCA-treated group. The histological study also showed that DNT had positive effect in treatment of NAFLD. CONCLUSION: DNT is an effective drug to treat patients with NAFLD of damp-heat syndrome type and is more effective than UDCA.

[Effects of Qinggan Huoxue Recipe and its decomposed formulas on CD14, Toll like receptor 4 and nuclear factor-kappaB expressed by Kupffer cells].

OBJECTIVE: To investigate the effects of Qinggan Huoxue Recipe (QGHXR), a compound Chinese herbal medicine, and its decomposed formulas Qinggan Recipe (QGR) and Huoxue Recipe (HXR) on expressions of CD14, Toll like receptor 4 (TLR(4)) and nuclear factor-kappaB (NF-kappaB) in Kupffer cells. METHODS: The isolated primary rat Kupffer cells were treated with lipopolysaccharide (LPS) for a certain period of time, a series of concentrations of drug-containing serums of QGHXR and its decomposed formulas were added, the expressions of NF-kappaB, tumor necrosis factor-alpha (TNF-alpha), CD14, and TLR(4) of the Kupffer cells were detected in different culture conditions by using Western blot, real-time reverse transcription polymerase chain reaction (RT-PCR) and ELISA methods respectively. RESULTS: QGR could down-regulate the expression of membrane receptor CD14, but the expression of NF-kappaB and TNF-alpha were not significantly decreased after QGR treatment. HXR could down-regulate the expression of membrane receptor TLR4 and inhibit the expressions of NF-kappaB and TNF-alpha. QGHXR could down regulate the expressions of membrane receptors CD14 and TLR(4) and inhibit the expressions of NF-kappaB and TNF-alpha. CONCLUSION: QGHXR can protect liver cells by down regulating the expressions of CD14, TLR(4) and NF-kappaB and inhibiting TNF-alpha expression.