Chang b Seo

Inhibitory effect of Yukmijihwang-tang, a traditional herbal formula against testosterone-induced benign prostatic hyperplasia in rats

BackgroundYukmijihwang-tang, a traditional herbal formula, has been used for treating disorder, diabetic mellitus and neurosis in China (Liu-wei-di-huang-tang in Chinese), Japan (Lokumijio-to in Japanese) and Korea for many years. In this study, we investigated the effects of Yukmijihwang-tang water extract (YJT) on the development of benign prostatic hyperplasia (BPH) using a rat model of testosterone propionate (TP)-induced BPH.MethodsA total of 30 rats were divided into five groups. One group was used as a control and the other groups received subcutaneous injections of TP for 4 weeks to induce BPH. YJT (200 or 400 mg/kg) was administered daily for 4 weeks to two groups by oral gavage concurrently with the TP. The animals were euthanized, the prostate and body weights were recorded, and tissues were subjected to hormone assays and histomorphology. In addition, we investigated proliferating cell nuclear antigen (PCNA) expression in the prostate using immunoblotting.ResultsAnimals with BPH showed significantly increased absolute and relative prostate weights, increased dihydrotestosterone levels in the serum or prostate and increased PCNA expression in the prostate; however, YJT-treated animals showed significant reductions compared with the animals with TP-induced BPH. Histomorphology also showed that YJT inhibited TP-induced prostatic hyperplasia.ConclusionsThese findings indicate that YJT effectively inhibited the development of BPH and might be a useful drug clinically.

An Extract of Crataegus pinnatifida Fruit Attenuates Airway Inflammation by Modulation of Matrix Metalloproteinase-9 in Ovalbumin Induced Asthma

Background Crataegus pinnatifida (Chinese hawthorn) has long been used as a herbal medicine in Asia and Europe. It has been used for the treatment of various cardiovascular diseases such as myocardial weakness, tachycardia, hypertension and arteriosclerosis. In this study, we investigated the anti-inflammatory effects of Crataegus pinnatifida ethanolic extracts (CPEE) on Th2-type cytokines, eosinophil infiltration, expression of matrix metalloproteinase (MMP)-9, and other factors, using an ovalbumin (OVA)-induced murine asthma model. Methods/Principal Finding Airways of OVA-sensitized mice exposed to OVA challenge developed eosinophilia, mucus hypersecretion and increased cytokine levels. CPEE was applied 1 h prior to OVA challenge. Mice were administered CPEE orally at doses of 100 and 200 mg/kg once daily on days 18–23. Bronchoalveolar lavage fluid (BALF) was collected 48 h after the final OVA challenge. Levels of interleukin (IL)-4 and IL-5 in BALF were measured using enzyme-linked immunosorbent (ELISA) assays. Lung tissue sections 4 µm in thickness were stained with Mayer’s hematoxylin and eosin for assessment of cell infiltration and mucus production with PAS staining, in conjunction with ELISA, and Western blot analyses for the expression of MMP-9, intercellular adhesion molecule (ICAM)-1 and vascular cell adhesion molecule (VCAM)-1 protein expression. CPEE significantly decreased the Th2 cytokines including IL-4 and IL-5 levels, reduced the number of inflammatory cells in BALF and airway hyperresponsiveness, suppressed the infiltration of eosinophil-rich inflammatory cells and mucus hypersecretion and reduced the expression of ICAM-1, VCAM-1 and MMP-9 and the activity of MMP-9 in lung tissue of OVA-challenged mice. Conclusions These results showed that CPEE can protect against allergic airway inflammation and can act as an MMP-9 modulator to induce a reduction in ICAM-1 and VCAM-1 expression. In conclusion, we strongly suggest the feasibility of CPEE as a therapeutic drug for allergic asthma.

Manassantin A inhibits cAMP-induced melanin production by down-regulating the gene expressions of MITF and tyrosinase in melanocytes

Abstract:  Microphthalmia‐associated transcription factor (MITF) is inducible in response to cAMP through the cAMP‐responsive element–binding protein (CREB) and plays a pivotal role in the melanocyte‐specific expression of tyrosinase or tyrosinase‐related proteins (TRPs) for melanin biosynthesis. Manassantin A from Saururus chinensis inhibits cAMP‐induced melanin production in B16 melanoma cells. Here, we focused on molecular basis of the antimelanogenic activity. Manassantin A consistently inhibited the cAMP elevator 3‐isobutyl‐1‐methylxanthine (IBMX)‐ or dibutyryl cAMP‐induced melanin production in B16 cells or in melan‐a melanocytes by down‐regulating the expression of tyrosinase or TRP1 gene. Moreover, manassantin A suppressed MITF induction through IBMX‐activated CREB pathway, directly inhibiting the Ser‐133 phosphorylation of CREB. However, manassantin A did not affect IBMX‐increased cAMP levels in these cells but also other cAMP‐dependent melanogenic pathways through post‐translational modifications of MITF. This putative molecular mechanism of manassantin A in the inhibition of melanin production suggests its pharmacological potential in skin hyperpigmentation.

Safety Evaluation of Yukmijihwang-tang: Assessment of Acute and Subchronic Toxicity in Rats

Yukmijihwang-tang (YMJ; Liu wei di huang tang (China), Rokumigan (Japan)) has been used in the treatment of diseases including renal disorder, cognitive vitality, and diabetes mellitus. However, there is very little information regarding the toxicity of YMJ to give an assurance of safety for clinical treatment. To provide safety information for YMJ, we evaluated its acute and sub-chronic toxicity in rats. The single-dose toxicity of YMJ was examined using Sprague-Dawley rats. Rats were treated with YMJ extract orally at 0, 500, 1000, or 2000 mg/kg body weight. After a single administration, clinical signs were observed every day for two weeks, and body weights were measured five times, including an initial measurement on day 1 (the day of administration). In the sub-chronic oral toxicity study, YMJ was administered to rats at 0, 500, 1000, or 2000 mg/kg/day for 13 weeks. Mortalities, clinical signs, body weight changes, food and water consumption, ophthalmologic findings, urinalysis, hematological and biochemical parameters, gross findings, organ weights, and histological examination were monitored during the study period. We found no mortality and no abnormalities in clinical signs, body weights, and necropsy findings for any of the animals in the acute and sub-chronic studies following oral administration in the rat at up to 2000 mg/kg/day YMJ. YMJ may not have any single-dose toxicity; the LD50 of YMJ was over 2000 mg/kg, and it is safe for rats. The no-observed-adverse-effect-level (NOAEL) was considered to be 2000 mg/kg/day.

Subacute toxicity and stability of Soshiho-tang, a traditional herbal formula, in Sprague–Dawley rats

BackgroudSoshiho-tang (SST, Xiao-chai-hu-tang in Chinese and Sho-saiko-to in Japanese), an oriental herbal formula, is used for treatment of chronic liver diseases. Although many researchers have studied the pharmacological properties of SST, information about its safety and toxicity is limited. Therefore, we evaluated the potential safety of SST in Sprague–Dawley rats over a period of 4-weeks.MethodsThe SST was administered once daily by gavage to male and female rats at doses of 0, 500, 1000 and 2000 mg/kg/day for 4 weeks. We measured the body weight, mortality, food consumption, ophthalmoscopy, urinalysis, hematology, serum biochemistry, gross pathological findings, absolute/relative organ weights and histopathology. In addition, we analyzed the component of SST and measured the stability of its component in SST according to study periods using high performance liquid chromatography.ResultsThe SST treatment did not result in any toxicologically significant changes in mortality, food consumption, ophthalmoscopy, urinalysis, hematology, serum biochemistry, gross pathological findings, absolute/relative organ weights and histopathology, except for salivation and reduction in body weight in the 2000 mg/kg/day male group. These findings in the 2000 mg/kg/day male group are considered toxicologically insignificant because they are not accompanied by other pathological findings, including in hematology, serum biochemistry and histopatholgy, and they do not exhibit a dose–response relationship. SST is detected three components including liquiritin, baicalin, and glycyrrhizin. In addition, there were not observed the significant differences among the contents of three components in SST according to storage periods.ConclusionThese results indicate that SST may be a safe material. Based on these results, the no-observed-adverse-effect level was more than 2000 mg/kg for both genders.

Extracts of Scutellariae Radix inhibit low-density lipoprotein oxidation and the lipopolysaccharide-induced macrophage inflammatory response

Traditional herbal formulas made from Scutellariae Radix (SR), the root of Scutellaria baicalensis, have previously been used in the treatment of inflammatory diseases, such as atherosclerosis. The aim of the present study was to investigate the effects of SR on low-density lipoprotein (LDL) oxidation and inflammation in macrophages, which are early events in the development of atherosclerosis. High-performance liquid chromatography photo-diode array analysis was used to obtain a three-dimensional chromatogram of SR. The antioxidative effects of SR were evaluated by determining its scavenging activities against ABTS and DPPH radicals. The inhibitory effect of SR on LDL oxidation was examined using a thiobarbituric acid-reactive substance assay and a relative electrophoretic mobility assay. In addition, the anti-inflammatory effects of SR were evaluated in lipopolysaccharide (LPS)-induced RAW264.7 murine macrophage cells. The results showed that SR exhibited radical-scavenging activities in a dose-dependent manner; in addition, SR attenuated the Cu2+-induced oxidation of LDL as well as significantly inhibited nitric oxide (NO) production and inducible NO synthase (iNOS) expression in LPS-induced RAW264.7 cells. Furthermore, SR induced the protein expression of heme oxygenase-1 (HO-1) in RAW264.7 cells. In conclusion, the results of the present study demonstrated that SR decreased the oxidation of LDL and suppressed inflammatory responses in macrophages, which occurred at least in part via the induction of HO‑1. These results therefore suggested that SR may be a potential therapeutic agent for the treatment of atherosclerosis.

Quantitative analysis and anti-inflammatory effects of Gleditsia sinensis thorns in RAW 264.7 macrophages and HaCaT keratinocytes

Gleditsia sinensis thorns have traditionally been used to treat edema and carbuncles and drain abscesses. In the present study, a simultaneous analysis of four flavonoids [(+)‑catechin, (‑)‑epicatechin, eriodictyol and quercetin] and two phenolic compounds (caffeic acid and ethyl gallate), obtained from a 70% ethanol extract of G. sinensis, was performed using high‑performance liquid chromatography‑photodiode array techniques. In addition, the inhibitory activities of the solvent fractions from a G. sinensis extract and its major constituents on the lipopolysaccharide‑stimulated production of inflammatory mediators by macrophage RAW 264.7 cells and the tumor necrosis factor (TNF)‑α and interferon (IFN)‑γ (TI)‑stimulated production of chemokines by HaCaT keratinocyte cells were investigated. The established analytical method showed high linearity, with a correlation coefficient of ≥0.9998. The limits of detection and quantification of the six compounds were 0.037‑0.425 and 0.124‑1.418 µg/ml, respectively. The ethyl acetate fraction inhibited nitric oxide and prostaglandin E2 production in RAW 264.7 cells and the production of thymus‑ and activation‑regulated chemokine (TARC) in HaCaT cells more than did the other fractions. Furthermore, the six compounds reduced the production of TARC, macrophage‑derived chemokine and regulated on activation normal T‑cell expressed and secreted in TI‑stimulated HaCaT cells; in particular, ethyl gallate and quercetin exhibited a significant dose‑dependent inhibition. Further elucidation of the signaling pathways involved in the T‑helper cell 2 chemokine inhibition by G. sinensis is necessary to facilitate the design of therapeutic agents for the inflammatory response.

Effect of saucerneol D on melanin production in cAMP-elevated melanocytes

Intracellular cAMP stimulates microphthalmia-associated transcription factor (MITF) induction in melanocytes through cAMP-responsive element binding protein (CREB), which plays a pivotal role in the gene expression of tyrosinase for melanin biosynthesis. In the present study, saucerneol D as a lignan constituent of Saururus chinensis (Saururaceae family) efficiently inhibited melanin production with IC50 values of 188–297 nM in B16 melanoma cells stimulated with α-melanocyte stimulating hormone (α-MSH) or other cAMP elevators. Moreover, saucerneol D down-regulated α-MSH-induced gene expression of tyrosinase at the transcription level in B16 cells, but it did not directly inhibit the catalytic activity of cell-free tyrosinase. As to the molecular basis of hypopigmenting action, saucerneol D inhibited α-MSH-induced phosphorylation of CREB in the cells, and sequentially suppressed MITF induction. Taken together, this study provides saucerneol D down-regulated the gene expression of tyrosinase, resulting in the inhibition of cAMP-induced melanin biosynthesis, and suggests pharmacological potential of the lignan structure in skin hyperpigmentation.

Anti-allergic effects of sesquiterpene lactones from the root of Aucklandia lappa Decne

Aucklandia lappa Decne, a well-known traditional herbal medicine, is used for the treatment of asthma, rheumatism, coughs, tuberculosis and numerous other diseases. The present study evaluated the inhibitory effects of the three sesquiterpene lactones costunolide, dehydrocostus lactone, and alantolactone, isolated from a 70% methanolic extract of Aucklandia lappa, on the expression of chemokine mRNA in HaCaT human keratinocyte cells. The cytotoxicities of the compounds on HaCaT cells were evaluated using a Cell Counting Kit8 assay. Furthermore, the inhibitory effects of the three compounds on chemokine expression in tumor necrosis factor (TNF)‑α‑ and interferon (IFN)‑γ‑stimulated HaCaT cells were analyzed by reverse transcription-polymerase chain reaction analysis. Treatment with the compounds caused a significant reduction in the mRNA expression of a range of chemokines, including TARC/CCL17, MDC/CCL22, RANTES/CCL5 and interleukin‑8 in TNF-α and IFN-γ-stimulated HaCaT cells. The present study indicated that costunolide, dehydrocostus lactone and alantolactone may have the potential to be used for treating inflammatory skin disorders by suppressing chemokine expression.

Safety assessment of So-cheong-ryong-tang: subchronic toxicity study in Crl:CD Sprague Dawley rats.

So-cheong-ryong-tang, an oriental herbal formula, is used in Korea for treating pulmonary disorders, including asthma, bronchitis and allergic diseases. The objective of the present study was to investigate the potential adverse effects, if any, of subchronic administration of So-cheong-ryong-tang aqueous extract (SCRT) in male and female rats. In the present study, 0, 1,000, 2,000 and 5,000 mg/kg/day of SCRT was administered to Crl:CD Sprague Dawley rats (10/gender/group) for 13 weeks via oral gavage. Administration of the SCRT did not result in any mortality. There were no clinical or ophthalmological signs, changes in urinalysis, body weight, food consumption, gross findings, hematology, serum biochemistry, organ weight or histopathology attributable to the administration of SCRT. Any alterations noted were incidental and consistent with those historically observed in the age and strain of rats used in the present study. Based on the results of the present study, the no observed adverse effect level for SCRT under the present experimental conditions was determined to be 5,000 mg/kg/day, the highest dose assessed, for both genders.