Formulation and evaluation of antioxidant and antityrosinase activity of Polygonum amplexicaule herbal gel.

Medicinal plants are long been used for pharmaceutical and cosmetic industry. Among medicinal plants, Polygonum amplexicaule of family polygonaceae has traditional use in medicines and skin care. P. amplexicaule belongs to genus Polygonum that contains several important phytochemicals and considered as a rich source of antioxidants. The present study was designed to formulate herbal gel containing P. amplexicaule extract and evaluate its different physical properties as well as antioxidants and antityrosinase activities. Chitosan gel base was used as gelling agent and different gel formulations were prepared by different concentrations of extracts and polymers. Physical properties like pH, colour, odour, appearance and homogeneity, spreadability, extrudability and stability were optimized and analysed. A stable gel formulation containing 1% chitosan gel base and 5% plant extract was prepared that showed good appearance and homogeneity, easily spread ability and excellent extrudability. This gel formulation was tested for antioxidant and skin whitening properties by DPPH free radical scavenging assay and tyrosinase inhibition assay respectively and ascorbic acid was used as reference standard. DPPH scavenging activity with an IC50 value of 0.446 mg/mL and tyrosinase inhibition activity with an IC50 value of 0.805 mg/mL was observed and results indicated that this herbal gel formulation has a good potential for cosmetic use.

Changes in immune responses of Helicoverpa armigera Hübner followed by feeding on Knotgrass, Polygonum persicaria agglutinin.

There is no study implying the effect of plant lectins on insect immune elements in both challenged and non-challenged conditions with entomopathogenic agents. Lectins may bind to immune receptors on the surface of insect hemocytes, thus inducing or even disabling common immune functions including hemocyte counts, nodulation/encapsulation, phenoloxidase activity, and synthesis of antimicrobial peptides. In the present study, effect of Polygonum persicaria L. agglutinin (PPA) on immune responses of Helicoverpa armigera Hübner was investigated by feeding artificial diet treated to the larvae. Subsequently hemocyte count and expression of some immune-related genes were considered for analyses. The two groups of larvae including control and PPA-treated (1%) were divided into four subgroups of intact, Tween-80 injected, latex-bead injected and Beauveria bassiana-injected. Except for intact larvae, the highest numbers of total and differential hemocyte counts were recorded 12 hr postinjection, however, the PPA-fed larvae showed a significantly lower hemocyte counts compared to control. The number of nodules in PPA-fed larvae was significantly lower than control, but the injected larvae of both control and PPA showed the highest nodulation 24 hr postinjection. Although the highest activity of phenoloxidase was observed 12 and 24 hr postinjection but its activity significantly decreased in PPA-fed larvae compared to control. Gene expression of antimicrobial peptides including attacin, cecropin, and peptidoglycan receptor proteins were significantly decreased in artificial diet-fed larvae containing PPA and then injected by B. bassiana spores and latex bead compared to control. These results clearly indicate adverse effects of PPA on immune responses in H. armigera.

Clinicopathological features of He Shou Wu-induced liver injury: This ancient anti-aging therapy is not liver-friendly.

BACKGROUND & AIMS: Polygonum Multiflorum Thumb (PMT), an ancient anti-aging Chinese herb known traditionally as He Shou Wu, has side effects of liver toxicity. To determine the main clinical and pathological characteristics of liver toxicity induced by PMT and the clinical course after its cessation. METHODS: Data of patients, diagnosed as drug-induced liver injury and hospitalised in Beijing Friendship Hospital from August 2005 to August 2017, were retrospectively reviewed. Clinical, pathological data and outcome after cessation of He Shou Wu were obtained and analysed. Kruskal-Wallis and Chi-square (χ2 ) tests were performed. RESULTS: Twenty-nine patients with He Shou Wu-induced liver injury were enrolled. The median age was 53 years (range 15-74) and 75.9% (22/29) were women. The most common symptom was jaundice (79.3%, 23/29). Of nine patients with liver biopsies, six showed acute cholestatic hepatitis, two acute, and one chronic hepatocellular injury pattern. The latency, liver chemistries and outcomes were comparable between pure He Shou Wu (5 patients) and its compounds (24 patients). Twenty-five of 29 patients (86.2%) had normal serum alanine aminotransferase levels after 45 days (range: 10-138 days) and total bilirubin of 46 days (range: 0-551 days). One patient was rechallenged with He Shou Wu and two developed autoimmune features. One patient died of liver failure and three had chronic persistent liver injury. CONCLUSIONS: The main clinicopathological injury pattern of He Shou Wu-induced liver injury is moderate to severe hepatitis with or without cholestasis. Most patients recover completely; however, chronic disease and death do occur.

[Chronic hepatotoxicity evaluation of Chinese medicinal herb Zishen Yutai pill prepared from Polygoni Multiflori Radix preparata in dogs].

To study the chronic hepatotoxicity of Chinese medicine Zishen Yutai pill (ZYP) prepared from Polygonum multiflorum with the recommended dosage in normal Beagle dogs. Low, middle and high doses of ZYP (1.5, 3.0, 6.0 g·kg⁻¹; i.e. 3×, 6× and 12× equivalent doses) were given orally to dogs for 39 consecutive weeks. At the same time, the same volume of deionized water was used as the solvent control group, one time a day. The general condition of the animals was observed every day during the period of administration, and the blood was collected before and 13, 26, 39, 43 weeks after administration to detect the biomarkers related to the hepatotoxicity of the dog serum. 2/7, 3/7 and 2/7 animals were dissected after 13, 39, and 43 weeks of administration to observe the pathological changes of the animal organs, weigh the mass of main organs and conduct pathological examination of the liver. As compared to the solvent control group, 11 liver hepatotoxicity traditional biomarkers such as ALT, AST were found no ZYP-related changes at month 3, 6, 9 of the administration and month 1 in recovery period; There was no significant difference in liver viscera index and liver pathology. Therefore, no obvious hepatotoxicity was shown by ZYP administered up to 6.0 g·kg⁻¹ for 9 months in normal dogs at doses of 1.5, 3.0, and 6.0 g·kg⁻¹.

Transcriptome analysis to assess the cholestatic hepatotoxicity induced by Polygoni Multiflori Radix: Up-regulation of key enzymes of cholesterol and bile acid biosynthesis.

Polygoni Multiflori Radix (PMR) has been commonly used as a tonic in China for centuries. However, PMR-associated hepatotoxicity is becoming a safety issue. Cholestasis often occurs in PMR-induced hepatotoxicity in clinical medicine, but the exact mechanism is not completely understood. An RNA-Seq method was employed, in the present study, to explore the molecular mechanism of cholestatic liver injury induced by PMR, characterized by the hepatic transcriptional response in rats exposed to 1 and 20 g/kg PMR for 90 days. Pathological changes seen in rat livers exposed to PMR included increased bile ducts in portal areas and biliary epithelial cell hyperplasia, which were accompanied by the elevation of serum biochemistries. Dose-dependent increases in the expression of 14 transcripts encoding enzymes involved in the cholesterol biosynthetic pathway were identified. Furthermore, cholesterol 7-alpha hydroxylase (Cyp7a1), a rate-limiting enzyme in the synthesis of bile acids (BAs) from cholesterol, was found to be upregulated by PMR treatment. Protein analysis by western blot suggested that expression of 3-hydroxy-3-methylglutaryl CoA reductase (Hmgcr) and Cyp7a1 were increased in a dose-dependent manner. Collectively, the present study demonstrates that PMR upregulates key enzymes for biosynthesis of cholesterol and BA, which poses the risk of cholestatic liver injury. SIGNIFICANCE: To the best of our knowledge, this is the first transcriptome analysis to highlight the main molecular changes occurring in rats chronic exposed to PMR. We have identified 39 specific differentially expressed genes (DEGs) that were present in various comparisons. A total of 14 of these altered gene transcripts were associated with cholesterol biosynthesis. Another factor of great importance in our opinion seemed to be the enhancement of bile acid (BA) biosynthesis, which were closely linked to cholesterol biosynthesis or metabolism. Our findings suggested that the disturbance on balance of BA formation and elimination might lead to a BA overload in hepatocytes, thereby resulting in liver injury.

[Dynamic serum metabolomics study of liver injury in rats caused by Polygonum multifulorum].

Liquid chromatography-mass spectrometry method was used for determination of metabolic fingerprint spectrum in rat serum, and the method of multivariate statistical analysis was used to compare the metabolism spectrum difference and screen significantly related biomarkers. The dynamic change trend was investigated at the same time. The dynamic metabonomics changes of liver injury in rats caused by Polygonum multifulorum(PM) were investigated; significantly related biomarkers were found and their dynamic change trend was investigated to provide basis for internal mechanism and early clinical diagnosis. There was certain difference in serum metabolic profile of the rats at different time points. Six potential biomarkers were screened through comparative analysis, including oleamide, lysoPC(16∶0), leukotriene A4, trans-tetra-dec-2-enoic acid, dihydrocortisol and 7a-hydroxydehydroepiandrosterone. These markers presented the dynamic change trend in the process of PM causing liver damage. The biomarkers contents had a significant change after one week of drug administration, more sensitive than ALT and AST. It can reveal the dynamic mechanism of PM causing liver damage and hepatic self-healing performance to some extent, with important application value and significance for monitoring liver function and early detecting diagnosis for patients with PM.

[Comparison of processed and crude Polygoni Multiflori Radix induced rat liver injury and screening for sensitive indicators].

To investigate the difference of liver injury in rats gavaged with crude and processed Polygoni Multiflori Radix. The 75% ethanol extract of crude and processed Polygoni Multiflori Radix (50 g · kg(-1) crude medicine weight/body weight) were continuous oral administered to rats for 6 weeks. Serum biochemical indicators were dynamically detected, the change of liver histopathology was assessed 6 weeks later. Principal component analysis (PCA) was adopted to screen sensitive indicator of the liver damage induced by polygoni multiflori radix. Biochemical tests showed that the crude Polygoni Multiflori Radix group had significant increase of serum ALT, AST, ALP, DBIL and TBIL (P < 0.01 or P < 0.05) and significant decreases of serum IBIL and TBA (P < 0.01 or P < 0.05), while the processed Polygoni Multiflori Radix group showed no obvious changes, compared to the untreated normal group. Histopathologic analysis revealed that crude Polygoni Multiflori Radix group exhibited significant inflammatory cells infiltration in portal area around the blood vessels, tissue destruction and local necrosis of liver cells. There were not obvious pathological changes in processed Polygoni Multiflori Radix group. The results demonstrated that the injury effect of processed Polygoni Multiflori Radix on liver injury of rats was significantly lower than that of unprocessed, and that processing can effectively reduce the hepatotoxicity of Polygoni Multiflori Radix. Traditional transaminase liver function indicators were not sensitive for crude Polygoni Multiflori Radix induced liver damage. The serum content of DBIL and TBIL can reflect the liver damage induced by crude Polygoni Multiflori Radix early and can be sensitive indicators for clinical monitoring the usage of it.

Effects of Polygonum multiflorum on the activity of cytochrome P450 isoforms in rats.

Polygonum multiflorum is a traditional Chinese medicinal herb used in clinical medicine to nourish the liver and kidney. However, in recent years, there have been increased reports of clinical adverse reactions associated with Polygonum multiflorum preparations, especially due to liver injury. The cocktail method can be used to assess the influence of Polygonum multiflorum on the activity of cytochrome P450 (CYP450) isoforms CYP2B6, CYP2C19, CYP2C9, CYP1A2, CYP3A4, and CYP2D6, which were reflected by changes in pharmacokinetic parameters in six specific probe drugs: bupropion, omeprazole, tolbutamide, phenacetin, midazolam, and metoprolol. Comprised the experimental rats were randomly divided into five groups: control group, alcohol extraction A group, alcohol extraction B group, water extraction A group, and water extraction B group. Each group five male rats and five female rats. Each of the groups received treatments by gavage as follows: control group was given normal saline, alcohol extraction A group was given 15 g/kg alcohol extract of Polygonum multiflorum (E15), alcohol extraction B group was given with 30 g/kg alcohol extract (E30), water extraction A group was given 15 g/kg water extract (W15), and water extraction B group was given 30 g/kg water extract (W30). The extract solution was orally administered once a day for 28 consecutive days. The mixture of six probe drugs was given by gavage, and blood samples were collected through the tail vein at different time points. Probe drug concentration in rat plasma was measured by liquid chromatography-mass spectrometry (LC-MS). In the treatment and control groups, Polygonum multiflorum alcoholic extract inhibited the activity of CYP2C19 and CYP2C9 and induced the activity of CYP1A2. Polygonum multiflorum aquous extract inhibited the activity of CYP2B6, CYP2C19, CYP2C9, CYP1A2, and CYP2D6. Pathological sections showed that in the alcohol extract group the liver was degenerated inconspicuously, and in the water extract group, the cytoplasm had vacuoles and particulate matter. The arrangement of liver cells was irregular.

[The idiosyncratic hepatotoxicity of Polygonum multiflorum based on endotoxin model].

The liver injury induced by Polygonum multiflorum Thunb. (PM) was investigated based on idiosyncratic hepatotoxicity model co-treated with lipopolysaccharide (LPS) at a non-hepatotoxic dose. Sprague-Dawley (SD) rats were intragastrically administered with three doses (18.9, 37.8, 75.6 g crude drug per kg body weight) of 50% alcohol extracts of PM alone or co-treated with non-toxic dose of LPS (2.8 mg·kg(-1)) via tail vein injection. The plasma alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities were assayed and the isolated livers were evaluated for histopathological changes. The dose-toxicity relationships of single treatment of PM or co-treatment of LPS were investigated comparatively to elucidate the idiosyncratic hepatotoxicity of PM. The results showed that no significant alterations of plasma ALT and AST activities were observed in the groups of solo-administration of LPS (2.8 mg·kg(-1), i.v.) or different dosage (18.9, 37.8 and 75.6 g·kg(-1), i.g.) of PM, compared to normal control group (P > 0.05); while significant elevations were observed in the co-administration groups of PM and LPS. Treatment with LPS alone caused slight infiltration of inflammatory cells in portal area but no evident hepatocytes injury. Co-treatment with LPS and PM (75.6 g·kg(-1), i.g.) caused hepatocyte focal necrosis, loss of central vein intima and a large number of inflammatory cell infiltration in portal areas. When further reduce the dosage of PM, significant increases of plasma ALT and AST activities (P < 0.05) were still observed in co-administration groups of LPS and PM (1.08 or 2.16 g·kg(-1)), but not in LPS or PM solo-administration groups. Nevertheless, the co-treatment of low dosage of PM (0.54 g·kg(-1)) with LPS did not induce any alteration of plasma ALT and AST. In conclusion, intragastric administration with 75.6 g·kg(-1) of PM did not induce liver injury in normal rats model; while the 2 folds of clinical equivalent dose of PM (1.08 g·kg(-1)) could result in liver injury in the LPS-based idiosyncratic hepatotoxicity model, which could be used to evaluate the idiosyncratic hepatotoxicity of PM.

Effect of sample preparation on components and liver toxicity of Polygonum multiflorum.

It was shown that different extracts had significant differences in the toxicity of Polygonum multiflorum. In this study, the effect of sample preparation on components and liver toxicity of different extracts from P. multiflorum were determined. Hepatoxic components were discovered based on biomembrane extraction. Comparative chemistry and toxicology between ethanol and water extracts were also performed. The results showed that ethanol extract had much stronger hepatotoxicity, the content of emodin-8-O-ß-d-glucopyranoside, physcion-8-O-ß-d-glucopyranoside, emodin and physcion was significantly higher in ethanol extract than in water extract, while the human hepatocytes extraction showed that 2,3,5,4'-tetrahydroxystilbene-2-O-ß-d-glucopyranoside, emodin-8-O-ß-d-glucopyranoside, physcion-8-O-ß-d-glucopyranoside, emodin and physcion had interaction with human hepatocytes. The hepatotoxic effect of these components was investigated on human hepatocytes LO2 cells and emodin-8-O-ß-d-glucopyranoside, physcion-8-O-ß-d-glucopyranoside, emodin and physcion were finally confirmed to be, at least partial, hepatotoxic components. The results showed that sample preparation has significant effect on components in extracts of P. multiflorum especially the components related to hepatotoxicity. Water extract, the conventional administration form of Chinese herbs, is prefer for phytotherapy before well understanding their chemistry and biological activities.

[Study on hepatotoxicity of aqueous extracts of Polygonum multiforum in rats after 28-day oral administration-analysis on correlation of cholestasis].

OBJECTIVE: To observe the liver injury degree of SD rats after 28-day administration of aqueous extracts of Polygonum multiflorum (AEPM) and the correlation with cholestasis mechanism. METHOD: Adult SD rats were orally administered with 30, 60 g x kg(-1) of APEM once every day for 28 d. After 28 d, the general condition of rats such as weight were observed, liver function-related indicators were detected. Bile was collected to determine total bile acid output, flow rate and density and changes in major compositions. Their livers were weighed then sent for histopathological examination. RESULT: AEPM did not change the general conditions and weights of rats. From the results of the related indicators of liver function and cholestasis, AEPM did not change the contents of ALT and AST in serum, but high dose of AEPM can increase the contents of ALP, GGT and TBA in serum (P < 0.05, P < 0.01) and decrease the content of TBIL in serum (P < 0.05). And the contents of GGT in serum of low dose rats were increased (P < 0.05). The bile flow was not changed by AEPM, but bile compositions of high dose male rats were obviously changed (TG increase, TBIL decrease, TBA decrease). The weights of liver and ratio of liver of the high dose rats were increased but showed no statistical significance. Pathologic examination displayed that there were only small pieces of necrosis in livers of several rats, without any severe disease. CONCLUSION: AEPM can obviously injure bile duct epithelial cells, intervene liver cell functions and change bile compositions in rats, thus it is proved to induce cholestasis without severe liver injury.

Toxicity of raw and processed roots of Polygonum multiflorum.

The roots of Polygonum multiflorum (Chinese name: He-Shou-Wu, HSW) are used in traditional Chinese medicine for many diseases in processed form or raw state. There are reports dealing with the toxicity of HSW. However, the toxicity is caused by over dosage or by the herb itself remains unclear. We evaluated the toxicity of raw and processed HSW on Kunming (KM) mice. For raw HSW, the toxicity of water decocta is much higher than that of acetone extract. Meanwhile, the toxicity of acetone extract of raw HSW is considerably higher than that of acetone extract of processed HSW. HPLC analyses revealed that the contents of characteristic compounds in raw HSW were changed after processing: the content of 2,3,4',5-tetrahydroxystilbene 2-O-ß-D-glucoside was decreased by 55.8%, whereas the content of emodin was increased by 34.0%. Thus, processing could reduce the toxicity of HSW. Thus, the toxicity of HSW does not depend on the content of anthranoid derivatives, it may be correlated with the content of tetrahydroxystilbene glucosides.

[Toxic hepatitis associated with Polygoni multiflori].

Toxic hepatitis has been reported as a major cause of acute hepatitis, but its potential induction by herbal remedies and/or health foods is usually neglected. We experienced a case of toxic hepatitis associated with Polygoni multiflori, a Chinese herb commonly known as Ho-Shou-Wu. A 54-year-old woman consumed Ho-Shou-Wu for 1 month, after which she experienced fatigue and overall weakness. A diagnosis of toxic hepatitis was made based on her clinical history, the findings for viral markers and other laboratory data, and ultrasonography. Her condition improved considerably after she stopped taking Ho-Shou-Wu. However, she resumed taking Ho-Shou-Wu immediately after discharge from hospital, which aggravated her symptoms and liver function. She was immediately readmitted and stopped taking Ho-Shou-Wu. Her relapse into hepatitis immediate after resuming consumption of the herb is strongly indicative of the validity of Koch's postulate in this case.

[Acute toxicity of Stephania cepharantha].

The acute toxicity of Stephania cepharantha was studied, LD50 of aqueous extract of its wet and dry root tuber oral were 41.4 g/kg and 22.9 g/kg respectively.