Protective effects of Liuweiwuling tablets on carbon tetrachloride-induced hepatic fibrosis in rats.

BACKGROUND: Liuweiwuling tablets (LWWL) are an herbal product that exerts remarkable effects on liver protection and aminotransferase levels, and they have been approved by the Chinese State Food and Drug Administration (CFDA). Clinical studies have found that LWWL can inhibit collagen production and reduce the levels of liver fibrosis markers in the serum. Thus, LWWL is expected to have beneficial effects in the treatment of liver fibrosis. The purpose of this study was to evaluate the pharmacological effects of LWWL. METHODS: Hepatic fibrosis was induced in rats via carbon tetrachloride (CCl4) treatment. The rats were treated twice weekly for 8 weeks with either 2 mL·kg- 1 body weight of a 50% solution of CCl4 in olive oil or olive oil alone by oral gavage. A subset of rats received daily intraperitoneal injections of either colchicine (0.2 mg/kg per day), LWWL (0.4, 1.6, or 6.4 g/kg per day), or vehicle (N = 12 for all groups) during weeks 9-12. The rats were sacrificed after 12 weeks. Pathological changes in hepatic tissue were examined using hematoxylin and eosin (H&E) and Sirius Red staining. Immunohistochemistry was performed to observe α-smooth muscle actin (α-SMA) and collagen type I (collagen I) protein expression. Western blotting was also used to detect α-SMA protein expression. Real-time quantitative reverse-transcription polymerase chain reaction (RT-qPCR) was used to detect transforming growth factor-1 (TGF-ß1), platelet-derived growth factor (PDGF), tissue inhibitor of metalloproteinase-1 (TIMP1), and tissue inhibitor of metalloproteinase-2 (TIMP2) mRNA expression. RESULTS: LWWL significantly reversed histological fibrosis and liver injury, reduced the hydroxyproline content in liver tissue, and decreased α-SMA and collagen I expression. LWWL also suppressed hepatic stellate cell (HSC) activation by reducing the expression of the profibrogenic factors TGF-ß1 and PDGF. The expression levels of TIMP1 and TIMP2, which regulate extracellular matrix (ECM) degradation, were decreased after CCl4 injury in LWWL-treated rats. CONCLUSIONS: These data suggest that LWWL may serve as a promising therapeutic agent to reduce fibrogenesis.

Liuweiwuling tablets attenuate BDL-induced hepatic fibrosis via modulation of TGF-ß/Smad and NF-κB signaling pathways.

ETHNOPHARMACOLOGICAL RELEVANCE: Liuweiwuling (LWWL) tablets contain a six-herb Chinese formula and are commonly prescribed to facilitate nourishment of the liver and kidneys, clear away toxic materials and activate blood circulation. Administration of LWWL is well known for its protective effects on the liver and its capacity to confer long-term stability in patients exhibiting reduced transaminase levels. Clinical studies have reported that LWWL can also be used for the treatment of liver fibrosis with associated treatment regimens resulting in a concomitant reduction in transforming growth factor ß1 (TGF-ß1) levels in the serum of patients with hepatic fibrosis. TGF-ß1 plays a prominent role in stimulating liver fibrogenesis and this effect is mediated by myofibroblasts (MFB) derived from hepatic stellate cells (HSCs). It is likely that this phenomenon underpins the antifibrotic effects associated with LWWL. AIM: The purpose of this study was to investigate the antifibrotic effects and mechanisms pertaining to LWWL. METHODS: Hepatic fibrosis was induced in rats following bile duct ligation (BDL). Rats that underwent BDL received daily gavage administration of colchicine (0.2mg/kg per day), LWWL (0.4, 1.6, 6.4g/kg per day) or PBS (for the control group). Pathological changes in hepatic tissue were examined using hematoxylin and eosin (HE) and sirius red staining. Immunohistochemical analysis was performed to monitor α-SMA and type I collagen (Collagen I) protein expression. Real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR) and Western blot analyses were used to monitor the expression of genes and proteins in the TGF-ß/Smad signaling pathway, including TGF-ß1, bone morphogenic protein and activin membrane-bound inhibitor (Bambi), Smad3, phosphorylated Smad3 (p-Smad3) and Smad7. We also monitored the expression of genes and proteins in the nuclear factor-κB (NF-κB) signaling pathway, including NF-κB p65, IκBα and phosphorylation of IκBα (p-IκBα), tumor necrosis factor-alpha (TNF-α), interleukin 6 (IL-6) and interleukin 1ß (IL-1ß). RESULTS: LWWL dose-dependently inhibited BDL-induced liver injury and hepatic fibrosis in rats. Furthermore, LWWL reduced liver tissue collagen deposition, hydroxyproline content, liver dysfunction and α-SMA expression in BDL-induced hepatic fibrosis rats. Moreover, LWWL markedly prevented activation of the TGF-ß/Smad signaling pathway by inhibiting expression of Smad2/3 and phosphorylation of Smad3, and upregulating the expression of Bambi and Smad7. In addition, LWWL regulated the expression of the inflammatory cytokines IL-1ß, TNF-α and IL-6 by inhibiting the activation of NF-κB p65 and the phosphorylation of IκBα, and increasing the expression of IκBα. CONCLUSIONS: LWWL can attenuate BDL-induced hepatic fibrosis in rats, and this effect may be due to modulation of the NF-κB-dependent inflammatory response and activation of HSC and TGF-ß/Smad-mediated synthesis and degradation of Collagen I.

Protective effect of Liuweiwuling Tablets on idiosyncratic liver injury induced by Polygonum multiflorum in rats based on immunomodulation / 中草药

Objective: To investigate the protective effect of Liuweiwuling Tablets of Polygonum multiflorum (PM)-induced idiosyncratic hepatotoxicity. Methods: Eighty Sprague-Dawley (SD) rats were randomly divided into control group, LPS group, PM group, LPS + PM group, LPS + PM + Liuweiwuling Tablets low, medium, and high dose (0.4, 0.8, and1.6 g/kg) groups, and LPS + PM + positive drug Biphenyl group (2 mg/kg). All groups were orally given Liuweiwuling Tablets and Biphenyl once daily for consecutive 2 d according to the different dosages. On day 3, except the normal group and LPS group oral administration of distilled water, all groups were given PM (crude drug 2.16 g/kg), 3 h after the group by the tail vein injection LPS (2.8 mg/kg) according to the different dosage; After 7 h, the rats were anesthetized with sodium pentobarbital, and the inferior vena cava was used to collect blood and liver tissue samples were collected. HE staining was used to observe the pathological changes of liver tissue, and the contents of ALT, AST, TNF-α, IL-1β, IL-6, and IFN-γ in rat plasma were determined. TUNEL assay analyzed hepatocyte apoptosis, and immunohistochemical staining was performed to detect the liver tissue expression activity of NF-κB p65. Results: Liuweiwuling Tablets could decrease ALT and AST levels of PM-induced idiosyncratic liver injury rat plasma, reduce liver pathological injury and hepatocyte apoptosis, inhibit the expression of NF-κB p65, and decrease plasma TNF-α and other inflammatory cytokines. Conclusion: Liuweiwuling Tablets can reduce inflammation through the inhibition of NF-κB signaling pathway, and prevent of PM induced idiosyncratic liver injury.

The protection effects of Liuweiwuling Tablets against concanavalin A-induced acute immunological liver injury in mice / 中国药理学通报

Aim To explore the protective effects and underlying mechanisms of Liu weiwuling Tablets (LW-WL)in concanavalin A (ConA)induced acute immu-nological liver injury in mice.Methods Mice were randomly divided into control,model,Bicyclol,LW-WL low dose (8 g·kg-1 )and LWWL high dose (16 g ·kg-1 )group.The medicattion was performed once daily for seven consecutive days,then the model of im-munological liver injury was prepared by intravenous injection of ConA (15mg·kg-1)in the tail of mice in each group except for the control group one hour after the last treatment.The pathological changes of liver tissues of mice were evaluated by HE staining with, and the levels of alanine amino transferase (ALT),as-partate aminotransferase (AST),and total bilirubin (TBIL)in serum were analyzed by colorimetric meth-od;the level of interleukin 12 (IL-12 ),interferon-γ(IFN-γ),tumor necrosis factor-α(TNF-α),interleu-kin 4 (IL-4)and interleukin 10 (IL-10)in liver was measured by real-time quantitative polymerase chain reaction (RT-qPCR);the changes of Th1 (IFN-γ) and Th2 (IL-4)cells were observed by flow cytometric (FCM)analysis;the expression of Th1/Th2 transcrip-tion factor T-bet/GATA-3 in liver tissue was detected by Western blot.Results Compared with normal con-trol group,the serum ALT,AST and TBIL were signif-icantly increased in model group, the pathological damage of the liver tissue was severe,and the necrosis and apoptosis of hepatic cells were large, which showed that the model was successful .Compared with model group,both low and high dose of LWWL could significantly reduce ALT,AST,TBIL levels in serum induced by ConA;Th1 cells in the spleen decreased, while Th2 cells increased;the expressions of IL-12, IFN-γand TNF-αmRNA were significantly inhibited with IL-4 and IL-10 mRNA expression elevated in mouse liver tissue;the expression of GATA-3 protein was up-regulated,T-bet protein expression showing no significant changes.Conclusion LWWL could regu-late Th1/Th2 balance,thus inhibiting the acute immu-nity hepatic injury induced by ConA.

Systematic evaluation of Liuweiwuling Tablets for treatment of liver fibrosis resulting from chronic hepatitis B / 中草药

Objective: To systematicly evaluate the efficacy and safety of Liuweiwuling Tablets combined with nucleotide drugs for the treatment of liver fibrosis resulting from chronic hepatitis B. Methods: Major databases (such as Pubmed, Embase, Cochrane Library (Issue 9, 2015), Chinese Biomedical Database (CBM), China National Knowledge Infrastructure (CNKI), VIP, and WanFang Data were searched from their inception to September 9, 2015 to collect comprehensively randomized controlled trials (RCT) of Liuweiwuling Tablets combined with nucleotide drugs on the treatment of liver fibrosis resulting from chronic hepatitis B. After two reviewers separately screened literature according to the inclusion and exclusion criteria, extracted data, and assessed the methodological quality of the included studies, data were analyzed with RevMan 5.3 software. Results: The total seven RCTs were included. Among all 1 021 patients involved, 529 cases were in the treatment group, while the other 492 cases were in the control group. The findings showed that compared with using nucleotide drugs alone for antiviral treatment, Liuweiwuling Tablets combined with nucleotide drugs were more efficiently in downregulating serum liver fibrosis index HA [SMD = -1.26, 95% CI (-1.92, -0.59), P = 0.000 2], LN [SMD = -1.55, 95% CI (-2.27, -0.83), P < 0.000 1], PC-III [SMD = -1.39, 95% CI (-2.21, -0.58), P = 0.000 8], IV-C [SMD = -1.49, 95% CI (-2.25, -0.72), P = 0.0001]; improved the liver function ALT [SMD = -1.17, 95% CI (-1.41, -0.94), P < 0.000 01], AST [SMD = -1.25, 95% CI (-1.54, -0.96), P < 0.000 01], TBIL [SMD = -0.87, 95% CI (-1.21, -0.53), P < 0.000 01]; increased total clinical effective rate [OR = 4.59, 95% CI (2.05, 10.25), P = 0.000 2]. While no advantage was found in reducing the incidences of HBV DNA overcast [OR = 1.51, 95% CI (0.96, 2.39), P = 0.08]; No significant adverse reactions were reported. Conclusion: Liuweiwuling Tablets combined with nucleotide drugs could be effective in improving liver function, controlling liver fibrosis, and increasing total clinical effective rate while its antiviral efficacy approximate with using the nucleotide drugs alone, less adverse reaction, and better tolerability; Although in recent years, the quantity and quality of RCTs research have a rising trend year by year, but still need to be from the clinical design of scientific, the standardization of the report, and many other links to improve research level.

Liuweiwuling tablets attenuate acetaminophen-induced acute liver injury and promote liver regeneration in mice.

AIM: To explore the mechanism of protection against acetaminophen-induced acute liver injury by Liuweiwuling tablets. METHODS: Intraperitoneal injections of acetaminophen (250 mg/kg) were used to induce acute liver injury in male C57BL/6 mice. A total of 24 healthy mice were randomly assigned to two groups: an acute liver injury group (control group) and a Liuweiwuling tablet group. Mice were given Liuweiwuling tablets or a vehicle (PBS) orally prior to the administration of acetaminophen. Serum alanine aminotransferase (ALT) and aspartate aminotransaminase (AST) levels were measured at different time points within one week, and pathological examinations of liver tissues were performed 36 h after induction of acute liver injury. Serum inflammatory cytokines, such as high mobility group box protein B1 (HMGB1), tumor necrosis factor (TNF)-α and interleukin IL-1ß, were detected using an ELISA method according to the manufacturer's instructions. Hepatic morphological changes at 36 h were assessed by hematoxylin and eosin staining. Expression of proliferating cell nuclear antigen (PCNA) in liver tissue was determined by Western blot analysis. The mRNA levels of hepatocyte proliferation markers (PCNA, CyclinD1 and p21) were detected by real-time quantitative reverse transcription-polymerase chain reaction. RESULTS: The levels of ALT/AST in the Liuweiwuling tablet group were decreased significantly at 6, 12 and 24 h compared to that of the control group (654.38 ± 120.87 vs 1566.17 ± 421.64, 1154.18 ± 477.72 vs 4654.84 ± 913.71 and 935.13 ± 252.34 vs 4553.75 ± 727.37, P < 0.01). Serum HMGB1 levels at 6 and 12 h for the Liuweiwuling tablet group were significantly lower than those of the control group (23.49 ± 3.89 vs 58.6 ± 3.65, 61.62 ± 13.07 vs 27.32 ± 5.97, P < 0.01). Furthermore, serum TNF-α and IL-1ß levels at 12 h in the Liuweiwuling tablet group were also significantly lower than those of the control group (299.35 ± 50.61 vs 439.03 ± 63.59, 57.42 ± 12.98 vs 160.07 ± 49.87, P < 0.01). Centrilobular necrosis was evident in liver tissue of mice with acetaminophen-induced acute liver injury, but was almost abolished in the Liuweiwuling tablet group. The expression levels of PCNA and CyclinD1 were up-regulated in liver tissue in the Liuweiwuling tablet group (321.08 ± 32.87 vs 157.91 ± 21.52, 196.37 ± 25.39 vs 68.72 ± 11.27, P < 0.01); however, expression of p21 in liver tissue was down-regulated compared to that of the control group (40.26 ± 9.97 vs 138.24 ± 13.66, P < 0.01). CONCLUSION: Liuweiwuling tablets can attenuate acute liver injury by decreasing inflammatory cytokine (HMGB1, TNF-α and IL-1ß) levels and promoting liver regeneration.