Puerarin ameliorates experimental alcoholic liver injury by inhibition of endotoxin gut leakage, Kupffer cell activation, and endotoxin receptors expression.

Puerarin, an isoflavone component extracted from Kudzu (Pueraria lobata), has been demonstrated to alleviate alcohol-related disorders. Our study examined whether puerarin ameliorates chronic alcoholic liver injury through inhibition of endotoxin gut leakage, the subsequent Kupffer cell activation, and endotoxin receptors expression. Rats were provided with the Liber-DeCarli liquid diet for 8 weeks. Puerarin (90 mg/kg or 180 mg/kg daily) was orally administered from the beginning of the third week until the end of the experiment. Chronic alcohol intake caused increased serum alanine aminotransferase, aspartate aminotransferase, hepatic gamma-glutamyl transpeptidase, and triglyceride levels as well as fatty liver and neutrophil infiltration in hepatic lobules as determined by biochemical and histologic assays. A significant increase of liver tumor necrosis factor α was detected by enzyme-linked immunosorbent assay. These pathologic effects correlated with increased endotoxin level in portal vein and upregulated protein expression of hepatic CD68, lipopolysaccharide-binding protein, CD14, Toll-like receptor 2, and Toll-like receptor 4. Meanwhile, the intestinal microvilli were observed to be sparse, shortened, and irregularity in distribution under the transmission electron microscope in conjunction with the downregulated intestinal zonula occludens-1 protein expression. These hepatic pathologic changes were significantly inhibited in puerarin-treated animals as were the endotoxin levels and hepatic CD68 and endotoxin receptors. Moreover, the pathologic changes in intestinal microvillus and the decreased intestinal zonula occludens-1 were also ameliorated with puerarin treatment. These results thus demonstrate that puerarin inhibition of endotoxin gut leakage, Kupffer cell activation, and endotoxin receptors expression is involved in the alleviation of chronic alcoholic liver injury in rats.

[Diagnostic value of clinical indices in syndrome differentiation of chronic hepatitis B: an exploration based on receiver operating characteristic curves and stepwise discriminant analysis].

OBJECTIVE: To explore the diagnostic value of 75 commonly used clinical laboratory markers for differentiation of traditional Chinese medicine syndromes such as liver and gallbladder damp-heat and liver depression and spleen deficiency in patients with chronic hepatitis B. METHODS: A total of 422 patients with chronic hepatitis B (CHB) were enrolled, including 300 patients with damp-heat in liver and gallbladder syndrome, and 122 patients with liver depression and spleen deficiency syndrome. Seventy-five commonly used clinical markers were selected, including liver and kidney function, clotting function, the quantitative detection of hepatic B virus (HBV) markers, HBV-DNA, blood count, hormones levels, cellular immunity indicators, humoral immunity indicators, lipid panel, protein electrophoresis, alpha-fetoprotein and liver fibrosis indicators. Receiver operating characteristic (ROC) curve was used to detect the diagnostic efficiency of single differential indicators, and stepwise discriminant analysis model was used to explore the discrimination efficiency of differential indices between two TCM syndromes in CHB. RESULTS: The differential indices between two CHB Chinese syndromes were albumin, prothrombin time, immunoglobulin A, immunoglobulin M, blood urea nitrogen, blood uric acid, basophils, basophil percentage and mean platelet volume. The area under ROC curve (AUC) of these indices was between 0.42 and 0.62, and the total false positive rate of own validation of stepwise discriminant analysis model, which was established by differential indices combination, was 35.3%, and the jackknife total error rate was 35.3%. CONCLUSION: Neither single differential index nor multiple differential indices determinant models provided appropriate determination of the TCM syndromes of patients with chronic hepatitis B, suggesting that clinical indicators have limited value in determining traditional Chinese medicine syndromes.

Effects of Puerariae Radix Extract on Endotoxin Receptors and TNF-α Expression Induced by Gut-Derived Endotoxin in Chronic Alcoholic Liver Injury.

Kudzu (Pueraria lobata) is one of the earliest medicinal plants used to treat alcohol abuse in traditional Chinese medicine for more than a millennium. However, little is known about its effects on chronic alcoholic liver injury. Therefore, the present study observed the effects of puerariae radix extract (RPE) on chronic alcoholic liver injury as well as Kupffer cells (KCs) activation to release tumor necrosis factor alpha (TNF-α) induced by gut-derived endotoxin in rats and macrophage cell line. RPE was observed to alleviate the pathological changes and lipids deposition in liver tissues as well as the serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), and hepatic gamma-glutamyl transpeptidase (GGT) activity. Meanwhile, RPE inhibited KCs activation and subsequent hepatic TNF-α expression and downregulated the protein expression of endotoxin receptors, lipopolysaccharide binding protein (LBP), CD14, Toll-like receptor (TLR) 2, and TLR4 in chronic alcohol intake rats. Furthermore, an in vitro study showed that RPE inhibited the expression of TNF-α and endotoxin receptors, CD14 and TLR4, induced by LPS in RAW264.7 cells. In summary, this study demonstrated that RPE mitigated liver damage and lipid deposition induced by chronic alcohol intake in rats, as well as TNF-α release, protein expression of endotoxin receptors in vivo or in vitro.

[Analysis of major herbs in Chinese herbal formula Jianpi Huoxue Decoction for improving intestinal permeability based on uniform design].

OBJECTIVE: To investigate the herbal medicines which play a main role in Chinese herbal formula Jianpi Huoxue Decoction for improving intestinal permeability and protect alcohol-induced liver injury and intestine damage, and to explore the analysis method for the material base of pharmacological effects of the Chinese herbal compound. METHODS: Sprague Dawley rats were given Lieber-DeCarli ethanol liquid diet once daily for 6 weeks to induce alcoholic liver injury. In step one, U(17)(17(16)) table of uniform design was adopted to design the experiment and the eight herbs of Jianpi Huoxue Decoction were screened to seek the herbs which play the main role. Three and a half hours before the rats were killed, each rat was administered lipopolysaccharide once, then blood sample was collected from portal vein and endotoxin content in plasma was detected as the index of intestinal permeability. The data were analyzed by stepwise regression to find the herbal drugs which had the best effects and the compatibility ratio of these drugs. In step two, the rats with Lieber-DeCarli liquid diet-induced liver injury were divided into four groups to test and verify the results. RESULTS: According to the obtained regression equation, Rasux Paeonia Alba (Baishao), Rhizoma Alismatis (Zexie) and Fructus Schisandrae Chinensis (Wuweizi) were the main herbal drugs in Jianpi Huoxue Decoction in improving intestinal permeability, and the doses for rats were 1.33, 0.50 and 0.17 g/kg respectively. In the verification experiment, combination of Baishao, Zexie and Wuweizi significantly decreased the endotoxin level in plasma of rats with Lieber-DeCarli-induced liver injury and showed reliability. CONCLUSION: Baishao, Zexie and Wuweizi are the major herbs of Jianpi Huoxue Decoction for improving intestinal permeability. Uniform design is efficient in screening the major herbs or their optimal combination in a certain Chinese compound.