Evaluation of the hepatoprotective effect of combination between hinokiflavone and Glycyrrhizin against CCl4 induced toxicity in rats

Liver diseases are one of the fatal syndromes due to the vital role of the liver. Most of the effective treatment of liver conditions are of natural origin. Silymarin (SI) is the standard drug used for treatment of impaired liver functions. Two natural compounds possessing promising liver protection and with different chemical structures namely; the bioflavonoid hinokiflavone (HF) isolated from Junipers phoenicea family Cupressaceae and the sweet saponin Glycyrrhizin (GL) present in Glycyrrhiza glabra (liquorice) were selected for the current study. Since the two compounds are of different nature, they may act by different mechanisms and express synergistic effect. Combination of the two compounds using to dose levels were challenged with single doses of HF, GL and SI as well. The comparison was monitored via measuring serum biochemical parameters including, aspartate aminotransferase (AST), alanine aminotransferase (ALT), gamma glutamyltranspeptidase (GGT), alkaline phosphatase (ALP) and total bilirubin, tissue parameters such as MDA, NP-SH and TP, histopathological study using light and electron microscope. Protective effect on kidney was also monitored histopathologically and biochemically through observing the levels of LDH, creatinine, creatinine-kinase, urea and uric acid. The combinations of HF and GL showed protective effect more than the used single doses of HF and GL alone. However, SI was superior to the used combination in the two used doses in all the measured parameters. The liver and kidney cells appearance under normal and electron microscope showed that SI treated groups showed almost normal cells with slight toxic signs. Cells from group treated with the higher doses of the combination of HF and GL showed slight signs of intoxication under light and electron microscope indicating good level of protection. Although the combination of HF and GL expressed good protection in the higher dose, however, the combination did not exceed the protective effect of SI.     

Hepatoprotective properties of red betel (Piper crocatum Ruiz and Pav) leaves extract towards H2O2-induced HepG2 cells via anti-inflammatory,antinecrotic, antioxidant potency

BackgroundProlonged exposure of free radicals, or known as reactive oxygen species (ROS), in hepatic cells may cause oxidative stress. Without proper treatment, it can induce liver injury and fatal hepatic disease, including cirrhosis. Red betel (Piper crocatum Ruiz and Pav) is one of Indonesia’s medicinal plants that has been known to exhibit antioxidant, anti-inflammatory activities. This study aims to determine hepatoprotective effect of red betel leaves extract (RBLE) towards liver injury.MethodHydrogen peroxide-induced HepG2 cells were used as liver injury model·H₂O₂-induced HepG2 cells were treated with 25 µg/mL and 100 µg/mL RBLE. Several parameters were observed, including TNF-α level through ELISA; necrotic, apoptotic, dead, live cells; and ROS level through flow cytometry analysis; and GPX gene expression through qPCR.ResultThe study showed that treatment with RBLE were able to decrease TNF-α level; necrotic and death cells percentage; as well as ROS level. On the other hand, it were able to increase apoptotic and live cells percentage; as well as GPX gene expression. Low concentration (25 µg/mL) of RBLE treatment exhibited stronger anti-inflammatory activity as it was resulted in the lower TNF-α level and were able to switched hepatic cell death pathway from necrosis to apoptosis as shown by the shifted of apoptotic cells and necrotic cells percentage. This lead to lower death cells and ultimately improve live cells percentage. Meanwhile high concentration of RBLE (100 µg/mL) exhibited stronger antioxidant properties as indicated by lower ROS level and higher GPX gene expression.ConclusionOverall, this study was able to demonstrate hepatoprotective effect of RBLE towards liver injury model through its anti-inflammatory and antioxidant activities.     

山苦荬保肝活性部位筛选及其化学成分研究

目的:筛选山苦荬保肝活性部位并对其化学成分进行研究.方法:采用系统溶剂法,以70％乙醇对山苦荬药材进行提取,用不同溶剂对所得提取物进行萃取,分别得石油醚、乙酸乙酯、正丁醇部位和剩余水部位.以对乙酰氨基酚诱导人肝细胞HL-7702复制肝损伤细胞模型,采用MTT法检测上述各部位(40μg/mL,均以生药量计)对损伤细胞的保...     

黄皮酰胺的合成

根据推测的生源关系,以仿生路线合成了有保肝活性的植物成分黄皮酰胺。     

Hepatoprotective effects of lychee (Litchi chinensis Sonn.): a combination of antioxidant and anti-apoptotic activities

Aim of the study

Gimjeng and Chakapat lychee (Litchi chinensis Sonn.) were evaluated for hepatoprotective activity on CCl₄-induced hepatotoxicity in rats.

Materials and methods

Fruit pulp extracts of the lychees were examined for vitamin C, phenolic contents, anti-lipid peroxidation activity and hepatoprotective effect. Male Wistar albino rats were intraperitoneally injected (ip) with CCl₄ (2 ml/kg), then were orally administered (po) with silymarin (100 mg/kg), and Gimjeng or Chakapat extracts (100 and 500 mg/kg). After ten days, the rats were sacrificed and their livers were examined histopathologically and immunohistochemically. Their serum glutamate-pyruvate transaminase, glutamate-oxalate transaminase, and alkaline phosphatase activities were analyzed. Apoptotic activity of the livers was assessed quantitatively.

Results

The Gimjeng and Chakapat extracts showed the contents of vitamin C (1.2 ± 0.6 and 4.3 ± 0.1 mg/100 g) and phenolics like trans-cinnamic acid and pelargonidin-3-O-glucoside (9.80 ± 0.21 and 19.56 ± 0.4 mg GAE/g extract, respectively), and trolox equivalent antioxidant capacity (TEAC) values (11.64 and 9.09 g/mg trolox), respectively. The Gimjeng as compared to the Chakapat demonstrated a better antioxidant activity as revealed by anti-lipid peroxidation activity with the TEAC values. Administration of CCl₄ in rats elevated the serum GPT, GOT, and ALP level whereas silymarin, Gimjeng and Chakapat extracts prevented these increases significantly. Significant decrease of apoptotic cells together with restoration of morphological changes confirmed the hepatoprotective effect in the CCl₄-induced rats pretreated with the extracts.

Conclusion

Antioxidant properties of the Gimjeng and Chakapat lychees as evidenced by the vitamin C and phenolic compounds, anti-lipid peroxidation and anti-apoptosis could explain the hepatoprotective effects in CCl₄-induced hepatotoxicity.     

保肝药在预防与治疗抗结核药所致肝损伤中的利用分析

目的：了解保肝药在预防与治疗抗结核药所致的肝损伤中的应用现状及趋势。方法：对我院2005-2009年在预防和治疗抗结核药引起的肝损伤中应用的保肝药的品种、用药数量、销售金额、用药频度（DDDS）、日均费用（DDC）等进行统计、分析。结果：我院所应用的保肝药涉及24个品种,5年间销售金额增长了280％,总DDDs增长了195％;注射用还原型谷胱甘肽、硫普罗宁注射液连续5年进入销售金额的前3名;葡醛内酯片的DDDs连续5年排第1位,注射用还原型谷胱甘肽的DDDs增加了265％,中成药的DDDs增长了259％。结论：保肝药在预防和治疗抗结核药引起的肝损伤中的应用呈明显的上升趋势。     

The hepatoprotective effect and chemical constituents of total iridoids and xanthones extracted from Swertia mussotii Franch

Ethnopharmacological relevance

Total iridoids and xanthones (TIXS) were extracted from Swertia mussotii Franch, one of the most important eight Tibetan medicines in China, which was recorded in the book of Jingzhu Bencao and used for clinical treatment of cholestatic hepatitis for many years. Our aim was to study the hepatoprotective effect and chemical constituents of the TIXS.

Materials and methods

Crude extracts were prepared using 90% ethanol, and individual fractions were collected following HPD-300 macroporous resin column chromatography. HPLC/MS was applied to qualitatively and quantitatively analyze the TIXS. Then, the alpha-naphthylisot hiocyanate-induced liver damage model was used to assess the hepatoprotective effect of the TIXS.

Results

A total of 12 compounds were identified by the fingerprint chromatography of the TIXS, and swertiamarin and swertianolin were shown to be its two main components. Oral administration of the TIXS at a dose of 35, 70 or 140 mg kg⁻¹, swertiamarin at a dose of 20 mg kg⁻¹ or swertianolin at a dose of 20 mg kg⁻¹, for 7 days in mice significantly reduced the alpha-naphthylisot hiocyanate-induced levels of alanine aminotransferase, aspartate aminotransferase and the total and direct bilirubins, and increased the bile flow (P<0.01).

Conclusion

These findings suggest that the TIXS exhibits significant hepatoprotective effect in the liver damage model induced by alpha-naphthylisot hiocyanate. Its active constituents include swertiamarin and swertianolin.     

Carthamus red from Carthamus tinctorius L. exerts antioxidant and hepatoprotective effect against CCl4-induced liver damage in rats via the Nrf2 pathway

Ethnopharmacological relevance

Carthamus red isolated from safflower (Carthamus tinctorius L., a Chinese traditional medicine) is evaluated for antioxidant and hepatoprotective activity.

Materials and methods

Carthamus red was isolated from a Na₂CO₃ extract of safflower and its analysis was carried out by HPLC/MS. Acute toxicity study was determined and the antioxidant activity was investigated using various established in vitro systems. An in vivo study against CCl₄-induced liver injury was also conducted and compared with that of silymarin, a known hepatoprotective drug.

Results

Carthamus red did not show any toxicity and mortality up to 2000 mg/kg dose, and it showed strong antioxidant ability in vitro. In the in vivo study, carthamus red treatment lowered the serum levels of ALT, AST, ALP and total protein in liver damage rat models. Meanwhile, Nrf2, GSTα and NQO1 expressions were up-regulated at the protein level by carthamus red intervention. Additionally, the activities of antioxidant enzymes and level of GSH were elevated by carthamus red intervention, while the content of TBARS, which is an oxidative stress marker, was lessened. HE stain analysis showed that the condition of liver damage was mitigated.

Conclusion

This study demonstrates that carthamus red may serve as a candidate with strong a hepatoprotective effect and antioxidant activity in liver damage.     

Antihepatotoxic activity of a quantified Desmodium adscendens decoction and d-pinitol against chemically-induced liver damage in rats

Ethnopharmacological relevance

The isolation of d-pinitol (or 3-O-methyl-d-chiro-inositol) from an aqueous decoction of Desmodium adscendens (Fabaceae) leaves and twigs is reported. The protective and curative effect of this decoction, in which d-pinitol was quantified, and of pure d-pinitol, against chemically-induced liver damage in rats has been evaluated.

Materials and methods

Enzyme levels of aspartate transaminase (AST), alanine transaminase (ALT) and alkaline phosphatase (ALP), which are among the usual biomarkers for liver damage, were determined in serum samples of experimental animals. The protective effects against d-galactosamine-induced and ethanol-induced liver damage of a decoction of D. adscendens, quantified on its main constituent d-pinitol, was investigated in rats. In addition, the curative effects of pure d-pinitol and D. adscendens against chronic d-galactosamine-induced and acute acetaminophen-induced hepatotoxicity in rats was studied. Silymarin was used as positive control.

Results

In a first experiment evaluating the protective effect against acute d-galactosamine-induced liver damage in rats, a significant decrease of AST and ALT was observed for the D. adscendens decoction at a dose equivalent to 5 mg/kg/day and 20 mg/kg/day d-pinitol, as well as 20 mg/kg/day pure d-pinitol. With respect to chronic ethanol-induced liver damage in rats, the protective effects of D. adscendens at doses equivalent to 2 mg/kg/day and 10 mg/kg/day d-pinitol, were not observed for serum AST and ALT levels. However, with respect to reduced mortality of animals, statistical analysis showed a trend towards significance for the Desmodium group receiving a dose equivalent to 10 mg/kg/day d-pinitol, versus the untreated hepatotoxic animals. Additional experiments in rat models of acute acetaminophen-induced and chronic d-galactosamine-induced liver damage indicated that D. adscendens decoction and pure d-pinitol had no curative effect when given in a dose equivalent to 10 mg/kg/day d-pinitol, or up to 20 mg/kg/day as a pure compound daily, respectively.

Conclusions

The aqueous decoction of D. adscendens showed a protective effect in rats against liver damage induced by d-galactosamine and ethanol, and this effect is at least in part due to the presence of d-pinitol. However, no curative effect of D. adscendens decoction or d-pinitol on liver damage induced by the tested chemicals could be demonstrated.     

Effect of a new hepatoprotective agent, YH-439, on the hepatobiliary transport of organic cations (OCS): Selective inhibition of sinusoidal OCs uptake without influencing glucose uptake and canalicular OCs excretion

The effect of a new hepatoprotective agent, YH-439, on the hepatobiliary transport of a model organic cation (OC), TBuMA (tributylmethylammonium), was investigated. The area under the plasma concentration-time curve (AUC) from time zero to 4 h following iv administration of TBuMA (6.6 micromol/kg) was increased significantly when YH-439 in corn oil (300 mg/kg) was orally administered to rats 24 h prior to the experiment. Nevertheless, the cumulative biliary excretion of TBuMA remained unchanged. As a consequence, the apparent biliary clearance (CLb) of TBuMA was decreased significantly as a result of YH-439 pretreatment, consistent with the fact that the in vivo excretion clearance of TBuMA across the canalicular membrane (CLexc) was not changed by the pretreatment. The in vitro uptake of TBuMA into isolated hepatocytes was decreased by one half by the pretreatment, owing to a decrease in the apparent Vmax and CLlinear, but the Km for the process remained constant. Most interestingly, however, the sinusoidal uptake of glucose, a nutrient, into hepatocytes was not influenced by the pretreatment, suggesting the YH-439 pretreatment specifically impaired the sinusoidal uptake of OCs. Thus, the OC-specific inhibition of hepatic uptake, without influencing the uptake of glucose, a nutrient, appeared to be associated with the hepatoprotective activity of YH-439.