Mechanistic studies on the alleviation of ANIT-induced cholestatic liver injury by Polygala fallax Hemsl. polysaccharides.

ETHNOPHARMACOLOGICAL RELEVANCE: Polygala fallax Hemsl. is a traditional folk medicine commonly used by ethnic minorities in the Guangxi Zhuang Autonomous Region, and has a traditional application in the treatment of liver disease. Polygala fallax Hemsl. polysaccharides (PFPs) are of interest for their potential health benefits. AIM OF THIS STUDY: This study explored the impact of PFPs on a mouse model of cholestatic liver injury (CLI) induced by alpha-naphthyl isothiocyanate (ANIT), as well as the potential mechanisms. MATERIALS AND METHODS: A mouse CLI model was constructed using ANIT (80 mg/kg) and intervened with different doses of PFPs or ursodeoxycholic acid. Their serum biochemical indices, hepatic oxidative stress indices, and hepatic pathological characteristics were investigated. Then RNA sequencing was performed on liver tissues to identify differentially expressed genes and signaling pathways and to elucidate the mechanism of liver protection by PFPs. Finally, Quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting were used to verify the differentially expressed genes. RESULTS: Data analyses showed that PFPs reduced the levels of liver function-related biochemical indices, such as ALT, AST, AKP, TBA, DBIL, and TBIL. PFPs up-regulated the activities of SOD and GSH, down-regulated the contents of MDA, inhibited the release of IL-1ß, IL-6, and TNF-α, or promoted IL-10. Pathologic characterization of the liver revealed that PFPs reduced hepatocyte apoptosis or necrosis. The RNA sequencing indicated that the genes with differential expression were primarily enriched for the biosynthesis of primary bile acids, secretion or transportation of bile, the reactive oxygen species in chemical carcinogenesis, and the NF-kappa B signaling pathway. In addition, the results of qRT-PCR and Western blotting analysis were consistent with those of RNA sequencing analysis. CONCLUSIONS: In summary, this study showed that PFPs improved intrahepatic cholestasis and alleviated liver damage through the modulation of primary bile acid production, Control of protein expression related to bile secretion or transportation, decrease in inflammatory reactions, and inhibition of oxidative pressure. As a result, PFPs might offer a hopeful ethnic dietary approach for managing intrahepatic cholestasis.

The efficacy and mechanism of Angelica sinensis (Oliv.) Diels root aqueous extract based on RNA sequencing and 16S rDNA sequencing in alleviating polycystic ovary syndrome.

BACKGROUND: Polycystic ovary syndrome (PCOS) leads to persistent anovulation, hyperandrogenism, insulin resistance, and polycystic ovary, and is mainly characterized by menstrual disorders, and reproductive dysfunction. Angelica sinensis (Oliv.) Diels root has been used in many classical formulas of traditional Chinese medicine, and is commonly used to treat various gynecological diseases. PURPOSE: To investigate the protective effect of water extract of A. sinensis root (WEA) on PCOS rats, and the mechanism by RNA sequencing, and 16S rDNA sequencing. METHODS: The PCOS rat model was established by letrozole combined with high-fat diet (gavage; 2 months), and treated with WEA (gavage; 2 g/kg, 4 g/kg or 8 g/kg; 1 month). To evaluate the therapeutic effect of WEA on PCOS rats, vaginal smear, hematoxylin-eosin staining, and biochemical indicators detection were performed. The rat ovarian tissue was analyzed by RNA sequencing, and the results were verified by qRT-PCR, and Western blot. 16S rDNA sequencing was used to analyze the gut microbiota of rats. RESULTS: The results of the vaginal smear, and hematoxylin-eosin staining showed that WEA improved estrous cycle disorder, and ovarian tissue lesions. WEA (4 g/kg or 8 g/kg; 1 months) alleviated hormone disorders, insulin resistance, and dyslipidemia. RNA sequencing showed that WEA intervention significantly changed the expressions of 2756 genes, which were enriched in phosphatidylinositol3-kinase/phosphorylated protein kinase B (PI3K/AKT), peroxisome proliferator-activated receptor (PPAR), mitogen-activated protein kinase (MAPK), AMP-activated protein kinase (AMPK), and insulin signaling pathways. 16S rDNA sequencing found that WEA increased the species diversity of gut microbiota, and regulated the abundance of some microbiota (genus level: Dubosiella, Bifidobacterium, Coriobacteriaceae (UCG-002), and Treponema; species level: Bifidobacterium animalis, Lactobacillus murinus, and Lactobacillus johnsonii). CONCLUSION: WEA regulated hormone, and glycolipid metabolism disorders, thereby relieving the PCOS induced by letrozole combined with high-fat diet. The mechanism was related to the regulation of PI3K/AKT, PPAR, MAPK, AMPK, and insulin signaling pathways in ovarian tissues, and the maintenance of gut microbiota homeostasis. Clarifying the efficacy and mechanism of WEA in alleviating PCOS based on RNA sequencing and 16S rDNA sequencing will guide the more reasonable clinical use of WEA.

Ethanolic extract of Pereskia aculeata induces Anti-Inflammatory Responses through P38/MK2/TTP-mediated signaling pathway.

Pereskia aculeata Miller, a member of the Cactaceae family, is a plant with pharmacological potential due to its containing compounds with various biological activities, which include anti-inflammatory, anti-cancer and analgesic activities. In this study, we evaluated the anti-inflammatory effects of an ethanolic extract of P. aculeata Miller (EEPA) and the signalling pathways by which it exerts these effects. In vitro, EEPA inhibited the secretion of inflammatory factors NO, IL-6 and PGE2 in ipopolysaccharide-stimulated RAW264.7 macrophages (P<0.05). Treatment of RAW264.7 cells with EEPA also significantly decreased the levels of P-P38 and P-MK2, while upregulating the expression of TTP (P<0.05). In vivo anti-inflammatory activity assays revealed that EEPA reduced the degree of foot and joint swelling, the splenic index and the serum concentrations of TNF-α and IL-6 in in adjuvant-induced arthritis rats (P<0.05). Similarly, EEPA treatment of mice inhibited the acetic acid-induced exudation of Evans blue dye from peritoneal capillaries and significantly prolonged heat-stimulated pain response time (P<0.05). Taken together, these results suggest that EEPA exerts anti-inflammatory effects in vitro and in vivo. Thus, this study provides experimental and technical support for the development of a novel anti-inflammatory treatment based on P. aculeata Miller.

Optimizing heterotrophic nitrification process: The significance of demand-driven aeration and organic matter concentration.

Heterotrophic nitrification and aerobic denitrification (HNAD) sludge were successfully acclimated. The effects of organics and dissolved oxygen (DO) on nitrogen and phosphorus removal by the HNAD sludge were investigated. The nitrogen can be heterotrophically nitrified and denitrified in the sludge at a DO of 6 mg/L. The TOC/N (total organic carbon to nitrogen) ratio of 3 was found to result in removal efficiencies of over 88% for nitrogen and 99% for phosphorus. The use of demand-driven aeration with a TOC/N ratio of 1.7 improved nitrogen and phosphorus removal from 35.68% and 48.17% to 68% and 93%, respectively. The kinetics analysis generated an empirical formula, Ammonia oxidation rate = 0.08917·(TOC·Ammonia)0.329·Biomass0.342. The nitrogen, carbon, glycogen, and poly-ß-hydroxybutyric acid (PHB) metabolism pathways of HNAD sludge were constructed using the Kyoto Encyclopedia of Genes and Genomes (KEGG). The findings suggest that heterotrophic nitrification precedes aerobic denitrification, glycogen synthesis, and PHB synthesis.

Electrochemical iron production to enhance anaerobic membrane treatment of wastewater.

Although iron salts such as iron(III) chloride (FeCl3) have widespread application in wastewater treatment, safety concerns limit their use, due to the corrosive nature of concentrated solutions. This study demonstrates that local, electrochemical generation of iron is a viable alternative to the use of iron salts. Three laboratory systems with anaerobic membrane processes were set up to treat real wastewater; two systems used the production of either in-situ or ex-situ electrochemical iron (as Fe2+ and Fe2+(Fe3+)2O4, respectively), while the other system served as a control. These systems were operated for over one year to assess the impact of electrochemically produced iron on system performance. The results showed that dosing of electrochemical iron significantly reduced sulfide concentration in effluent and hydrogen sulfide content in biogas, and mitigated organics-based membrane fouling, all of which are critical issues inherently related to sustainability of anaerobic wastewater treatment. The electrochemical iron strategy can generate multiple benefits for wastewater management including increased removal efficiencies for total and volatile suspended solids, chemical oxygen demand and phosphorus. The rate of methane production also increased with electrochemically produced iron. Economic analysis revealed the viability of electrochemical iron with total cost reduced by one quarter to a third compared with using FeCl3. These benefits indicate that electrochemical iron dosing can greatly enhance the overall operation and performance of anaerobic membrane processes, and this particularly facilitates wastewater management in a decentralized scenario.

Marchantia polymorpha L. Flavonoids Protect Liver From CCl4-Induced Injury by Antioxidant and Gene-Regulatory Effects.

Marchantia polymorpha L. (MPL), a common type of liverwort, has been used as herbal medicine to improve liver function in China for many years. Although modern studies revealed that MPL contains various polyphenols, terpenoids, and bis[bibenzyls], its biological effects on liver function have never been systemically studied in any animal model. In this study, flavonoids were extracted from MPL and the components in the MPL flavonoids as well as the antioxidant capacity of MPL flavonoids were analysed. A rat model of liver injury was induced by intraperitoneal injection of 10% carbon tetrachloride (CCl4). MPL flavonoids were administered daily at a dose of 50, 100, and 200 mg/kg body weight to the rats for 2 weeks prior to injection of CC14. Treatment with MPL flavonoids, especially at a dose of 200 mg/kg, attenuated CCl4-induced increases in alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, γ-glutamyl transpeptidase, nitric oxide, malondialdehyde, tumour necrosis factor-α, interleukin-1ß, and interleukin-6, as well as reductions in superoxide dismutase and glutathione peroxidase. Microarray analyses showed that co-treatment with MPL flavonoids and CCl4 up-regulated many antioxidant and anti-apoptotic genes, but down-regulated several pro-inflammatory genes, compared to treatment with CCl4 alone. PCR and western blot assays further identified that MPL flavonoids increased GPX1, TMX1, TXN, and XIAP expression, but decreased IL-1 and IL1RAP expression and inhibited Jak/stat3 signalling. In conclusion, MPL flavonoids exerted hepatoprotective effects via antioxidant and gene regulatory mechanisms. (Altern Ther Health Med.

Phosphate recovery from the P-enriched brine of AnMBR-RO-IE treating municipal wastewater via an innovated phosphorus recovery batch reactor with nano-sorbents.

Municipal wastewater is a very unique pool full of energy and useful substances. Though the innovative integrated anaerobic membrane bioreactor and reverse osmosis-ion exchange (AnMBR-RO-IE) process can produce high-grade reclaimed water with high energy efficiency, phosphorus resources recovery in the WWTPs has been rarely reported thus far. This study evaluated the feasibility of a phosphorus recovery batch reactor (PRBR) as an approach for the phosphate production from the P-enriched brine from AnMBR-RO-IE. With operating PRBR for 162 cycles, high to 85% of P recovery rate was obtained for 145 cycles, leading to a P production rate of 6.17 g/m3 domestic wastewater with nano-sorbents (NSs) consumption rate of 10.2 g/m3. Acidification pretreatment efficiently improved the adsorption capacity and reduced the NSs renewing frequency. High adsorption selectivity of NSs contributed to low impurities (<0.3%) in the P-enriched reclaimed solution. Moreover, the integrated AnMBR-RO-IE-PRBR process saved 47% of energy consumption compared to the present NEWater production process in Singapore. The innovative PRBR reactor was competitive compared to the commonly-used chemical precipitation methods in conventional WWTPs in terms of phosphorus recovery/loss and energy balance. It is expected that the proposed integrated process can offer new insights into the direction of phosphorus reclamation in the future WWTPs.

The effects of Gentiana dahurica Fisch on alcoholic liver disease revealed by RNA sequencing.

ETHNOPHARMACOLOGICAL RELEVANCE: The root of Gentiana dahurica Fisch (called Qin-Jiao in China), a traditional Chinese medicine, is used in China to treat alcoholic liver disease (ALD), but there has been no scientific report on the treatment of ALD. AIM OF THE STUDY: To investigate the therapeutic effects of Gentiana dahurica Fisch ethanol extract (GDEE) on ALD and to reveal its possible mechanism of action using RNA sequencing. MATERIALS AND METHODS: The model of ALD was established by continuous gavage with alcohol in mice, and GDEE was used to treat ALD. Pathological observation (HE staining, oil red O staining) and biochemical indicators were performed to evaluate liver tissue lesions and efficacy of GDEE. RNA sequencing analysis of liver tissues was carried out to elucidate the pathogenesis of ALD and the mechanism of hepatoprotective effect by GDEE. The RNA sequencing results were verified by detecting mRNA and protein expressions of acetyl coenzyme A carboxylase α (Acacα), fatty acid synthase (Fasn) and carnitine palmitoyltransferase 1A (Cpt1a) by quantitative real-time polymerase chain reaction (PCR) and Western blot. RESULTS: Measurements of biochemical parameters showed that GDEE could inhibit the increased transaminase activities in the serum and lipid levels in the liver caused by alcohol. It was observed that GDEE could alleviate fatty degeneration, edema and cell necrosis caused by alcohol in the liver tissue. RNA sequencing analysis of liver tissues found that 719 genes and 1137 genes were significantly changed by alcohol and GDEE, respectively. GDEE reversed most of the changes in triglycerides synthesis-related genes up-regulated by alcohol. GDEE up-regulated most of the genes involved in the fatty acid degradation in ALD mice, while alcohol had little effect on them. In addition, GDEE suppressed most of the genes involved in cholesterol synthesis that were up-regulated by alcohol. GDEE up-regulated genes related to bile acid synthesis in ALD mice, and down-regulated genes related to bile acid reabsorption, while alcohol had no significant effect on genes related to bile acid metabolism. In the validation experiments, the Acacα, Fasn and Cpt1a expressions quantified by real-time PCR and Western blot were consistent with the RNA sequencing results. CONCLUSIONS: GDEE can alleviate liver damage and steatosis in ALD mice, and its mechanism of action may be related to the process of regulating triglycerides and cholesterol.

The effect of intermittent drying and wetting stormwater cycles on the nutrient removal performances of two vegetated biofiltration designs.

Vegetated biofiltration systems (biofilters) are now a well-established technology for treatment of urban stormwater, typically showing high nutrient uptake. However, the impact of high temporal variability of rainfall events (further exacerbated by climate change) on nitrogen and phosphorus removal processes, within different biofiltration designs, is still unknown. Hence, a laboratory-based study was conducted to uncover mechanisms behind nutrient removal in biofilters across different drying and wetting regimes. Two sets of experimental columns were based on (1) the standard biofiltration design (unsaturated zone only), and (2) combination of unsaturated and saturated (submerged) zone (SZ) with additional carbon source. Columns were watered with synthetic stormwater according to three drying and wetting schemes, exploring 1, 2, 3, 4 and 7-week drying. Hydraulic performance, soil moisture and pollutant removal were monitored. The results show that hydraulic conductivity of SZ design experiences less change over time compared to standard design, due to slower media drying, crack formation and lower plant die-off. Varied drying lengths challenged both designs differently, with 2-week drying resulting in significant drop of performance across most pollutants in standard design (except ammonia), while SZ design was able to retain high performance for up to four weeks of drying, sustaining microbial and plant uptake. Increased oxygenation of SZ columns during short-term drying was beneficial for ammonia and phosphorus removal. While SZ design showed better performance and quicker recovery for nitrogen removal, in regions with inter-rain event shorter than two weeks, the standard design (no saturated zone, no carbon source) can achieve similar if not better results.

Study on the protective effect and mechanism of Dicliptera chinensis (L.) Juss (Acanthaceae) polysaccharide on immune liver injury induced by LPS.

The purpose of this study is to use Dicliptera chinensis (L.) Juss (Acanthaceae) polysaccharide (DCP) to act on the NF-κB inflammatory pathway and Fas/FasL ligand system, in order to find a new method to improve immune liver injury. Lipopolysaccharide (LPS) was used to establish an injury model in vivo (Kunming mice) and in vitro (LO2 cells). In this experiment, hematoxylin-eosin (H&E) staining and related biochemical indicators were used to observe the pathological changes of liver tissues, oxidative stress and inflammatory reactions. Immunohistochemistry, ELISA, RT-PCR and Western blot were used to detect protein or mRNA expressions associated with inflammation response and apoptosis. The experimental results show that the model group has obvious liver cell damage and inflammatory infiltration. After DCP intervention, it could significantly reduce the levels of ALT, AST, ALP, TBIL and MDA in serum, and increase the content of SOD and GSH-Px. In addition, DCP can reduce the expression level of NF-κB in the liver and reduce the release of downstream inflammatory factors TNF-α, IL-6 and IL-1ß, thereby reducing the inflammation. At the same time, DCP can significantly inhibit the expression of Fas/FasL ligand system and apoptosis related-proteins and mRNA, which in turn can reduce cell apoptosis. In conclusion, DCP can alleviate liver injury by inhibiting liver inflammation and apoptosis, which provides a new strategy for clinical treatment of immune liver injury.

Mechanism of phosphate adsorption on superparamagnetic microparticles modified with transitional elements: Experimental observation and computational modelling.

Microparticle sorbents (MPSs) containing nano-Fe3O4 core, nano-layered double hydroxides shell modified with different transitional elements (TE-MPSs) are effective for capturing phosphate from wastewater. In present article, different TE-MPSs loaded with Ce, La, Zr and Hf were synthesized and the phosphorous removal performances were compared. The molecular-level mechanism of phosphate adsorption was successfully simulated by pseudo-atom model based on virtual crystal approximation method, which was in line with the spectroscopic data. The results confirmed the inter-sphere complexation reaction between phosphate and TE-MPSs under neutral pH. And the adsorption mechanism probably changed during cycles of adsorption/regeneration. As revealed by elemental composition analysis, ion exchange mechanism probably played an important part in phosphate adsorption in Cycle 1 while the ligand complexation became dominant after regenerated with NaOH in the following cycles. This work indicates that the highly effective, easily separable and repeatedly usable TE-MPSs has the potential for phosphate sequestration and recovery from sewage wastewater.

Hepatoprotective effects of Dicliptera chinensis polysaccharides on dimethylnitrosamine-induced hepatic fibrosis rats and its underlying mechanism.

ETHNOPHARMACOLOGICAL RELEVANCE: Dicliptera chinensis is a traditional herbal medicine used anciently in China for hepatopathy treatment, especially in south areas. Our several studies have demonstrated that dicliptera chinensis polysaccharides (DCP), which has a markedly protective effects on chemistry-induced models of acute liver injury in rats. In this study, we further investigated the potentially hepatoprotective effect of dicliptera chinensis polysaccharides (DCP) on hepatic fibrosis (HF) rats induced by dimethylnitrosamine (DMN). MATERIAL AND METHODS: The 96 rats were randomly divided into six groups (n=16, per group), the normal control group intragastrically administrated normal saline, model control group intraperitoneally injected with 0.5% DMN solution at 1.6mL per kg (three times a week); colchicine intragastrically administrated group (0.2mgkg(-)(1)d(-1))+DMN-treated rats; DCP intragastrically administrated groups (100mgkg(-)(1)d(-)(1), 200mgkg(-1)d(-1), 300mgkg(-1)d(-1))+DMN-treated rats. At the end of 8 weeks, all rats were sacrificed. RESULTS: Pathological examination showed that high and medium doses of DCP presented remarkable effect in ameliorating hepatic fibrosis, alleviate the inflammation, necrosis and reduced collagen deposits. DCP effectively improved the liver function, as revealed in being lowered sero-enzyme levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBIL) while increased albumin (ALB), and being reduced sero-concentrations of tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6) in the HF rats. Additionally, the contents of hyaluronic acid (HA), collagen type Ⅳ (Ⅳ-C), type III precollagen (PCIII) and laminin (LN) in the hepatic tissue of HF rats were markedly decreased, whereas the expressions of transforming growth factor-ß l (TGF-ß l), collagen type I (Col- I), metal protease-1 (TIMP-1), nuclear factor-kappa B (NF-κB) expression in the hepatic tissue were notably down-regulated. CONCLUSION: DCP exerts effectively antagonistic activity on DMN-caused hepatotoxicity in HF rats, which the anti-fibrotic mechanisms are associated with regulating functionally serous enzymes, improving metabolic function and inhibiting inflammatory reaction in liver tissue.

Model development for prediction of soil water dynamics in plant production.

Optimizing water use in agriculture and medicinal plants is crucially important worldwide. Soil sensor-controlled irrigation systems are increasingly becoming available. However it is questionable whether irrigation scheduling based on soil measurements in the top soil could make best use of water for deep-rooted crops. In this study a mechanistic model was employed to investigate water extraction by a deep-rooted cabbage crop from the soil profile throughout crop growth. The model accounts all key processes governing water dynamics in the soil-plant-atmosphere system. Results show that the subsoil provides a significant proportion of the seasonal transpiration, about a third of water transpired over the whole growing season. This suggests that soil water in the entire root zone should be taken into consideration in irrigation scheduling, and for sensor-controlled irrigation systems sensors in the subsoil are essential for detecting soil water status for deep-rooted crops.

Surrogates for herbicide removal in stormwater biofilters.

Real time monitoring of suitable surrogate parameters are critical to the validation of any water treatment processes, and is of particularly high importance for validation of natural stormwater treatment systems. In this study, potential surrogates for herbicide removal in stormwater biofilters (also known as stormwater bio-retention or rain-gardens) were assessed using field challenge tests and matched laboratory column experiments. Differential UV absorbance at 254mn (ΔUVA254), total phosphorus (ΔTP), dissolved phosphorus (ΔDP), total nitrogen (ΔTN), ammonia (ΔNH3), nitrate and nitrite (ΔNO3+NO2), dissolved organic carbon (ΔDOC) and total suspended solids (ΔTSS) were compared with glyphosate, atrazine, simazine and prometryn removal rates. The influence of different challenge conditions on the performance of each surrogate was studied. Differential TP was significantly and linearly related to glyphosate reduction (R(2) = 0.75-0.98, P < 0.01), while ΔTP and ΔUVA254 were linearly correlated (R(2) = 0.44-0.84, P < 0.05) to the reduction of triazines (atrazine, simazine and prometryn) in both field and laboratory tests. The performance of ΔTP and ΔUVA254 as surrogates for herbicides were reliable under normal and challenge dry conditions, but weaker correlations were observed under challenge wet conditions. Of those tested, ΔTP is the most promising surrogate for glyphosate removal and ΔUVA254 is a suitable surrogate for triazines removal in stormwater biofilters.

Anti-diabetic effects of puerarin, isolated from Pueraria lobata (Willd.), on streptozotocin-diabetogenic mice through promoting insulin expression and ameliorating metabolic function.

Diabetes is a chronic disease characterized by the metabolic disorder in specific tissues. Our present study was designed to assess the potential benefits of puerarin (PR) on hypoglycemic and hypolipemic effects in diabetic mice induced by streptozotocin (STZ). The results achieved from these experiments showed that glycemia in STZ-diabetogenic mice were significantly reduced following the PR administration, while serum insulin concentration was increased. In addition, PR contributed to improving the dyslipidemia conditions. Histopathological examination indicated that the STZ-lesioned pancreas tissue in PR-administrated mice was effectively alleviated. Meanwhile, intrapancreatic protein levels of insulin receptor substrate-1 (IRS-1) and insulin-like growth factor-1 (IGF-1) were up-regulated, respectively. On the other hand, endogenous mRNA levels of skeletal muscle insulin receptor (InsR) and peroxisome proliferators-activated receptor α (PPARα) were increased after administration of PR. Taken together, these findings reveal that puerarin effectively exerts the hypoglycemic and hypolipemic roles, which its potential anti-diabetic activity is associated with elevating insulin expression and maintaining metabolic homoeostasis in STZ-diabetogenic mice.

Puerarin, isolated from Pueraria lobata (Willd.), protects against hepatotoxicity via specific inhibition of the TGF-ß1/Smad signaling pathway, thereby leading to anti-fibrotic effect.

Recently, the TGF-ß1/Smad signaling pathway has been investigated in the pathogenesis of hepatofibrosis, and pharmacological treatment of liver fibrosis targeted this pathway to determine its contribution to the inhibition of fibrotic development. Importantly, ethnopharmacology-derived Pueraria lobata has been reported to effectively reverse the fibrotic process in the liver. In the present study, we performed dimethylnitrosamine (DMN)-induced liver fibrosis in rats to assess the benefits of puerarin (PR), which was isolated from Pueraria lobata (Willd.), on ECM-derived hepatocytes associated with the TGF-ß1/Smad pathway. Our results showed that the serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), hyaluronic acid (HA), laminin (LN), type III precollagen (PCIII) and type IV collagen (CIV) were significantly reduced by PR treatment, while hepatic homogenates showed decreased levels of hydroxyproline (Hyp) and collagen I (Col I). Masson's trichrome staining indicated that the DMN-induced liver fibrosis was alleviated. In addition, the protein expression levels of transforming growth factor-ß l (TGF-ß l), smad2, smad3, α-SMA and TIMP-1 were downregulated specifically by PR treatment, whereas the protein expression levels of smad7 and MMP-1 were upregulated. Furthermore, we evaluated the PR-mediated inhibitory effect on TGF-ß1-treated proliferation and activation in a rat liver stellate cell line (HSC-T6). These data resulted in inhibition of the cell growth of HSC-T6 in a dose-dependent manner and a reduction in TßRI, smad2 and smad3 expressed proteins in the presence of PR on TGF-ß1-treated HSC-T6 cells, while smad7 levels were downregulated. Taken together, these findings identify a unique effect for PR-regulation of the TGF-ß1/Smad pathway in blocking fibrotic development and provide a promising strategy for hepatofibrosis treatment.

Puerarin, isolated from Kudzu root (Willd.), attenuates hepatocellular cytotoxicity and regulates the GSK-3ß/NF-κB pathway for exerting the hepatoprotection against chronic alcohol-induced liver injury in rats.

Puerarin (PR) has been utilized as a phytomedicine to managing liver disease in China. Thus, this study aimed to evaluate the potential PR-mediated hepatoprotective role against chronic alcohol-induced liver injury in rats. The results indicated that serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP) and pro-inflammatory cytokines were significantly reduced following PR treatment, while the albumin (ALB) level was increased. Meanwhile, intrahepatic contents of alcohol dehydrogenase (ADH), aldehyde dehydrogenase (ALDH) were elevated. Pathological examination showed that alcohol-lesioned hepatocytes were mitigated through the PR treatment. In addition, the endogenous levels of glycogen synthase kinase-3ß (GSK-3ß) at the protein level and ß-catenin expression at the mRNA level were notably down-regulated, whereas the tumor necrosis factor alpha (TNF-α) and nuclear factor-kappa B (NF-κB) proteins in the liver tissue were effectively decreased following the PR treatment. Together, these findings demonstrate that PR mediates hepatoprotection against alcohol-induced liver injury. The mechanisms underlying the cytoprotective effects of PR are associated with inhibiting immunotoxicity in hepatocytes and regulating the GSK-3ß/NF-κB pathway, thereby maintaining metabolic homeostasis in the liver tissue.

Puerarin improves metabolic function leading to hepatoprotective effects in chronic alcohol-induced liver injury in rats.

Puerarin (PR), an active component extracted from the kudzu root, has been widely used as an ethno-medicine to treat hepatopathy in China. Therefore, the aim of the present study was to investigate the hepatoprotective action of PR in chronic alcohol-induced liver injury in rats. Data showed that the serum levels of alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) were elevated following PR administration. In addition, the levels of endogenous CYP2E1, CYP1A2, and CYP3A proteins in liver tissue were also gradually decreased following PR treatment. Histopathological examinations suggested that alcohol-induced hepatocellular lesions were mitigated by PR treatment. Collectively, these data indicate that PR contributes to cytoprotection against alcohol-induced liver lesions through improving metabolic function.

[Research on active extracts of Dicliptera chinensis on liver protection].

OBJECTIVE: To search pharmacody namic active extracts of Dicliptera chinensis for protecting liver. METHOD: Rats cute liver injury model was induced by CCl4 to screen the liver protection of four different fraction extracted from an ethanol extract of D. chinensis. RESULT: The n-butanol fraction and water fraction can prevent the increase of the ALT, AST and MDA (P < 0.01, P < 0. 05), and increase the SOD compared to the normal groups (P < 0.05). CONCLUSION: The results show that the pharmacody namic active extracts of D. chinensis are the n-butanol and water fractions.

Cloning, characterization and TBT exposure response of CuZn superoxide dismutase from Haliotis diversicolor supertexta.

The full-length cDNA and genomic DNA of a cytoplasmic copper, zinc superoxide dismutase (CuZn-sod) were cloned from the hepatopancreas of small abalone Haliotis diversicolor supertexta by RT-PCR, RACE and TAIL PCR. The full-length cytoplasmic CuZn-sod cDNA (designated sasod) comprises 984 bp. Its ORF encodes a polypeptide of 154 amino acids with a predicted molecular mass of 15.7 kDa and theoretical isoelectric point of 6.30. The deduced amino acid (designated saSOD) shares a common consensus pattern with the SODs of vertebrate and invertebrate animals. The full-length sasod genomic DNA comprises 5,574 bp, containing five exons and four introns. The splice donor and acceptor sequence of the four introns is 5'GT-AG3'. Real time quantitative PCR analysis revealed that sasod expression level in hepatopancreas of small abalone was no significant difference at 2, 6, 48 and 192 h post TBT exposure (P > 0.05). However, the sasod expression level at 12 and 24 h post TBT exposure was decreased significantly (P < 0.05).