Promotion of Hair Growth by Rosmarinus officinalis Leaf Extract

Topical administration of Rosmarinus officinalis leaf extract (RO‐ext, 2 mg/day/mouse) improved hair regrowth in C57BL/6NCrSlc mice that experienced hair regrowth interruption induced by testosterone treatment. In addition, RO‐ext promoted hair growth in C3H/He mice that had their dorsal areas shaved. To investigate the antiandrogenic activity mechanism of RO‐ext, we focused on inhibition of testosterone 5α‐reductase, which is well recognized as one of the most effective strategies for the treatment of androgenic alopecia. RO‐ext showed inhibitory activity of 82.4% and 94.6% at 200 and 500 µg/mL, respectively. As an active constituent of 5α‐reductase inhibition, 12‐methoxycarnosic acid was identified with activity‐guided fractionation. In addition, the extract of R. officinalis and 12‐methoxycarnosic acid inhibited androgen‐dependent proliferation of LNCaP cells as 64.5% and 66.7% at 5 µg/mL and 5 μM, respectively. These results suggest that they inhibit the binding of dihydrotestosterone to androgen receptors. Consequently, RO‐ext is a promising crude drug for hair growth. Copyright © 2012 John Wiley & Sons, Ltd.     

红花水提物对乳鼠心肌细胞H_2O_2所致损伤的保护作用及ESR谱研究

目的:观察红花水提物对体外培养大鼠乳鼠心肌细胞H2O2损伤的保护作用及电子自旋共振(ESR)波谱的影响。方法:建立心肌细胞H2O2损伤模型,实验分5组,空白对照组,模型对照组:H2O2(1 mmol.L-1)孵育1 h;红花水提物500,100,20 mg.L-1剂量组:分别给予不同浓度的红花水提物,预处理24 h,然后换为H2O2(1 mmol.L-1)孵育1 h。实验结束后分别收集细胞上清液测定乳酸脱氢酶(LDH)、丙二醛(MDA)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)、一氧化氮(NO)、一氧化氮合酶(NOS)和黄嘌呤氧化酶(XOD)活性,用自旋捕集技术结合电子自旋共振波谱技术(Electron Spin Res-onance,简称ESR)检测各组细胞悬液中氧自由基信号的强弱。结果:红花水提物各剂量组能够明显提高H2O2损伤后的心肌细胞的搏动频率,使细胞存活率提高,减少心肌细胞LDH的释放,降低培养液中MDA和XOD的含量,同时提高SOD,NO和GSH-Px的活性,使细胞悬液中的ESR氧自由基信号明显减弱。结论:红花水提物对体外培养乳鼠心肌细胞具有保护作用,其机制与其减少活性氧产生、增强氧自由基的清除、增强内源性抗氧化酶的活性有关。     

Anti‐Nociceptive Effect of Resveratrol During Inflammatory Hyperalgesia via Differential Regulation of pro‐Inflammatory Mediators

Sensitization of nociceptive neurons by inflammatory mediators leads to hypersensitivity for normal painful stimuli which is termed hyperalgesia. Oxidative stress is an essential factor in pathological pain; therefore, antioxidants qualify as potential anti‐hyperalgesic agents. The present study examines the efficacy of the natural antioxidant resveratrol in complete Freund's adjuvant (CFA) induced hyperalgesic rats. Thermal hyperalgesia was measured at different time points by paw withdrawal latency test and confirmed by c‐Fos expression in spinal dorsal horn. The impact of resveratrol treatment on inflammatory mediators at peripheral (paw skin) and central (spinal cord) sites was determined during early (6 h) as well as late phase (48 h) of hyperalgesia. Intraplanter injection of CFA increased the level of cytokines IL‐1β, TNF‐α and IL‐6 as well as inflammatory enzymes COX‐2 and iNOS in paw skin in both phases. In case of spinal cord, the level of COX‐2 was found to be elevated in both phases, whereas iNOS could not be detected. The cytokines were found to be elevated only in late phase in spinal cord. Administration of resveratrol (20 mg/kg) shifted the level of all inflammatory mediators towards normal, except cytokines in paw skin. The present study suggests that the anti‐nociceptive effect of resveratrol is implicated at both peripheral and central sites in a tissue specific manner. Copyright © 2016 John Wiley & Sons, Ltd.     

Hydroxysafflor Yellow A Attenuates Bleomycin‐induced Pulmonary Fibrosis in Mice

Hydroxysafflor yellow A (HSYA) is an active component of Carthamus tinctorius L., and we want to investigate whether HSYA attenuates pulmonary fibrosis induced by bleomycin (BLM) in mice. The mice received a BLM via oropharyngeal aspiration, and HSYA was intraperitoneally injected. Arterial blood gas analysis was performed. Morphological changes and hydroxyproline content were measured. mRNA expression of transforming growth factor‐β1 (TGF‐β1), connective tissue growth factor, α‐smooth muscle actin (α‐SMA), and collagen I was measured by real‐time polymerase chain reaction. α‐SMA‐positive cells in lung tissues were detected by immunohistochemical staining. A549 cell was cultured, and morphological changes were observed after TGF‐β1 and HSYA treatment. mRNA expression was detected by real‐time polymerase chain reaction. Phosphorylation of Smad3 was evaluated by western blotting. HSYA decreased the lung consolidation area and collagen deposition in mice with pulmonary fibrosis. The blood gas changes due to BLM were attenuated by HSYA. HSYA also alleviated the BLM‐induced increase of TGF‐β1, connective tissue growth factor, α‐SMA, and collagen I mRNA levels. HSYA treatment inhibited the increase of α‐SMA expression, Smad3 phosphorylation, the morphological changes in lung tissue. HSYA inhibits Smad3 phosphorylation and elevated expression of collagen I mRNA in epithelial–mesenchymal transition induced by TGF‐β1. Copyright © 2016 John Wiley & Sons, Ltd.     

银杏生发合剂促进小鼠毛发生长的作用研究

目的:观察银杏生发合剂对毛发生长的促进作用。方法:取C57BL/6小鼠,随机分为章光101组,银杏合剂高、中、低剂量组,将小鼠背部双侧脱发,左侧分别外擦章光101育发剂,银杏生发合剂167.7mg/ml、100.6mg/ml、33.5 mg/ml,右侧外涂30%乙醇溶剂,每日2次。观察21天内动物新生毛发生长情况,测定各组动物新生毛发长度及重量,采用ELISA法测定皮肤组织中血管内皮生长因子及Ⅰ型胶原含量。结果:银杏生发合剂高剂量能显著增加动物毛发生长速度(P<0.05),中剂量能显著增加动物新生毛发长度及皮肤中血管内皮生长因子含量(P<0.05)。结论:外擦银杏生发合剂有显著的促生发作用,并能增加皮肤中VEGF的分泌。     

Astragaloside III activates TACE/ADAM17‐dependent anti‐inflammatory and growth factor signaling in endothelial cells in a p38‐dependent fashion

Astragaloside III (AS‐III) is a triterpenoid saponin contained in Astragali Radix and has potent anti‐inflammatory effects on vascular endothelial cells; however, underlying mechanisms are unclear. In this study, we provided the first piece of evidence that AS‐III induced phosphorylation of TNF‐α converting enzyme (TACE) at Thr735 and enhanced its sheddase activity. As a result, AS‐III reduced surface TNFR1 level and increased content of sTNFR1 in the culture media, leading to the inhibition of NF‐κB signaling pathway and attenuation of downstream cytokine gene expression. Furthermore, AS‐III induced TACE‐dependent epidermal growth factor receptor (EGFR) transactivation and activation of downstream ERK1/2 and AKT pathways. Finally, AS‐III induced activation of p38. Both TACE activation and EGFR transactivation induced by AS‐III were significantly inhibited by p38 inhibitor SB203580. Taken together, we concluded that AS‐III activates TACE‐dependent anti‐inflammatory and growth factor signaling in vascular endothelial cells in a p38‐dependent fashion, which may contribute to its cardiovascular protective effect.     

Praeruptorin a inhibits lipopolysaccharide‐induced inflammatory response in murine macrophages through inhibition of NF‐κB pathway activation

Praeruptorin A (PA) is a pyranocoumarin compound isolated from the dried root of Peucedanum praeruptorum Dunn (Umbelliferae). However, the antiinflammatory effect of PA has not been reported. The present study investigated the antiinflammatory effect of PA in lipopolysaccharide (LPS)‐stimulated RAW 264.7 macrophage cells. PA significantly inhibited the LPS‐induced production of nitric oxide (NO), interleukin‐1β (IL‐1β) and tumor necrosis factor‐α (TNF‐α). The mRNA and protein expressions of inducible nitric oxide synthase (iNOS), IL‐1β and TNF‐α were also suppressed by this compound. Further study showed that PA decreased the cytoplasmic loss of inhibitor κB‐α (IκB‐α) protein and inhibited the translocation of NF‐κB from cytoplasm to nucleus. Taken together, the results suggest that PA may exert antiinflammatory effects in vitro in LPS‐stimulated RAW 264.7 macrophages through inhibition of NF‐κB signal pathway activation. Copyright © 2010 John Wiley & Sons, Ltd.     

A Polyphenol‐rich Pomegranate Fruit Extract Suppresses NF‐κB and IL‐6 Expression by Blocking the Activation of IKKβ and NIK in Primary Human Chondrocytes

Pomegranate fruit extract (PE) rich in polyphenols has been shown to exert chondroprotective effects, but the mechanism is not established. Here, we used an in vitro model of inflammation in osteoarthritis (OA) to investigate the potential of PE to suppress interleukin 1 beta (IL‐1β)‐stimulated expression of inflammatory cytokine IL‐6, generation of reactive oxygen species (ROS) levels, and investigated the mechanism of NF‐κB inhibition by analyzing the activation of the kinases upstream of IκBα in primary human chondrocytes. Total and phosphorylated forms of kinases and expression of IL‐6 were determined at protein and mRNA levels by western immunoblotting and Taqman assay, respectively. Dihydrorhodamine 123 staining estimated ROS generation. Pomegranate fruit extract inhibited the mRNA and protein expression of IL‐6, generation of ROS, and inhibited the IL‐1β‐mediated phosphorylation of inhibitor of nuclear factor kappa‐B kinase subunit beta (IKKβ), expression of IKKβ mRNA, degradation of IκBα, and activation and nuclear translocation of NF‐κB/p65 in human chondrocytes. Importantly, phosphorylation of NF‐κB‐inducing kinase was blocked by PE in IL‐1β‐treated human OA chondrocytes. Taken together, these data suggest that PE exerts the chondroprotective effect(s) by suppressing the production of IL‐6 and ROS levels. Inhibition of NF‐κB activation by PE was blocked via modulation of activation of upstream kinases in human OA chondrocytes. Copyright © 2017 John Wiley & Sons, Ltd.     

Hydroxy‐Safflower Yellow A inhibits the TNFR1‐Mediated Classical NF‐κB Pathway by Inducing Shedding of TNFR1

Hydroxy‐safflower yellow A (HSYA) is the major active component of safflower, a traditional Asia herbal medicine well known for its cardiovascular protective activities. The purpose of this study was to investigate the effect of HSYA on TNF‐α‐induced inflammatory responses in arterial endothelial cells (AECs) and to explore the mechanisms involved. The results showed that HSYA suppressed the up‐regulation of ICAM‐1 expression in TNF‐α‐stimulated AECs in a dose‐dependent manner. High concentration (120 μM) HSYA significantly inhibited the TNF‐α‐induced adhesion of RAW264.7 cells to AECs. HSYA blocked the TNFR1‐mediated phosphorylation and degradation of IκBα and also prevented the nuclear translocation of NF‐κB p65. Moreover, HSYA reduced the cell surface level of TNFR1 and increased the content of sTNFR1 in the culture media. TNF‐α processing inhibitor‐0 (TAPI‐0) prevented the HSYA inhibition of TNFR1‐induced IκBα degradation, implying the occurrence of TNFR1 shedding. Furthermore, HSYA induced phosphorylation of TNF‐α converting enzyme (TACE) at threonine 735, which is thought to be required for its activation. Conclusively, HSYA suppressed TNF‐α‐induced inflammatory responses in AECs, at least in part by inhibiting the TNFR1‐mediated classical NF‐κB pathway. TACE‐mediated TNFR1 shedding can be involved in this effect. Our study provides new evidence for the antiinflammatory and anti‐atherosclerotic effects of HSYA. Copyright © 2016 John Wiley & Sons, Ltd.     

Protective Effects of Fermented Citrus Unshiu Peel Extract against Ultraviolet‐A‐induced Photoageing in Human Dermal Fibrobolasts

The aqueous extracts of Citrus unshiu peel containing flavonoid glycosides was used as co‐substrate with Schizophyllum commune mycelia producing β‐glucosidase and its biological activities were studied. β‐glucosidase‐produced S. commune mycelia converted the glycosides (narirutin and hesperidin) into aglycones (naringenin and hesperetin). The photoprotective potential of fermented C. unshiu peel extract with S. commune (S‐CPE) was tested in human dermal fibroblasts (HDFs) exposed to UVA. It was revealed that S‐CPE had an inhibitory effect on human interstitial collagenase (matrix metalloproteinase, MMP‐1) expression in UVA‐irradiated HDFs. The treatment of UVA‐irradiated HDFs with S‐CPE resulted in a dose‐dependent decrease in the expression level of MMP‐1 mRNA. The UVA irradiation raised the proportion of senescence‐associated β‐galactosidase (SA‐β‐gal) positive cells in comparison with the normal control group. The treatment of UVA‐irradiated HDFs with S‐CPE was shown to decrease the level of SA‐β‐gal (by approximately 45% at an S‐CPE concentration 0.1%, w/v) compared with the UVA‐irradiated HDFs. It was found that S‐CPE containing hesperetin has notable collagen biosynthetic activity for fibroblasts, indicating that S‐CPE can be promising cosmetic ingredients. Copyright © 2012 John Wiley & Sons, Ltd.     

Study on antifibrotic effects of curcumin in rat hepatic stellate cells

Suppression of activation or fibrogenesis and induction of apoptosis, in hepatic stellate cells (HSCs) have been proposed as therapeutic strategies against liver fibrosis. Curcumin, an active compound isolated from yellow curry pigment of turmeric (Curcuma longa Linn), has been demonstrated to be an effective anti‐inflammatory and antioxidant compound. In this study, we investigated the in vitro antifibrogenic effects of curcumin on HSCs at the concentration range of (1–40 µM). A cell line of rat HSCs (HSC‐T6) was stimulated with transforming growth factor‐β1 (TGF‐β1). The inhibitory effects of curcumin (1.25～10 µM) on fibrosis‐related markers including α‐smooth muscle actin (α‐SMA) and collagen were assessed. In addition, the induction effects of curcumin (20～40 µM) on apoptosis in HSC‐T6 cells were also assessed by Hoechst and propidium iodide stains. Curcumin (1.25～10 µM) concentration‐dependently suppressed TGF‐β1‐induced α‐SMA expression and collagen deposition in HSC‐T6 cells, without cytotoxicity. Whereas, higher concentrations of curcumin (20～40 µM) induced cell apoptosis and cytochrome c release in HSC‐T6 cells. Our results suggest that curcumin exerted antifibrotic effects, possibly through two different mechanisms depending on its concentrations. At lower concentrations (1.25～10 µM), curcumin exerted antifibrogenic effects, whereas at higher concentrations (20～40 µM), curcumin exerted induction of apoptosis in HSCs. Copyright © 2009 John Wiley & Sons, Ltd.     

Neuroprotective Effect of Vitamin E Isoforms Against Chronic Alcohol‐induced Peripheral Neurotoxicity: Possible Involvement of Oxidative‐Nitrodative Stress

Small‐fiber painful peripheral neuropathy is one of the long‐term complications of alcohol for which there is no reliable successful therapy available. The precise mechanisms by which chronic alcohol consumption produces peripheral nerve fiber damage and loss remain unclear. Emerging data from clinical and preclinical studies suggest that increased oxidative‐nitrodative stress mediated release of proinflammatory cytokines from damaged neural tissues may play a central role in the pathogenesis of alcoholic neuropathy. The present study investigated the effect of both the isoforms of vitamin E (α‐tocopherol and tocotrienol) against chronic alcohol‐induced peripheral neuropathy in rats. Ethanol treated rats showed a significant decrease in paw‐withdrawal threshold in both Randall‐Selitto and von‐Frey hair tests along with a significant reduction in tail flick latency in the tail‐immersion test. A decreased pain threshold was associated with significant alterations in oxidative‐nitrodative stress markers and an increase in proinflammatory cytokines (TNF‐α and IL‐1β). The 4‐week treatment with tocotrienol significantly ameliorated behavioral, biochemical and molecular alterations in alcohol treated rats. However, α‐tocopherol failed to produce any protective effect. The results of the present study suggest that oxidative‐nitrodative stress mediated cytokine signaling may be responsible for alcohol‐induced peripheral neurotoxicity and tocotrienol treatment might be beneficial in chronic alcoholics exhibiting neuropathy. Copyright © 2012 John Wiley & Sons, Ltd.     

Phloroglucinol Protects INS‐1 Pancreatic β‐cells Against Glucotoxicity‐Induced Apoptosis

Decreasing numbers, and impaired function, of pancreatic β‐cells are key factors in the development of type 2 diabetes. This study was designed to investigate whether phloroglucinol protected pancreatic β‐cells against glucotoxicity‐induced apoptosis using a rat insulinoma cell line (INS‐1). High glucose treatment (30 mM) induced INS‐1 cell death; however, the level of glucose‐induced apoptosis was significantly reduced in cells treated with 100‐μM phloroglucinol. Treatment with 10–100‐μM phloroglucinol increased cell viability and decreased intracellular levels of reactive oxygen species, nitric oxide, and lipid peroxidation dose‐dependently in INS‐1 cells pretreated with high glucose. Furthermore, phloroglucinol treatment markedly reduced the protein expression of Bax, cytochrome c, and caspase 9, while increasing anti‐apoptotic Bcl‐2 protein expression. Cell death type was examined using annexin V/propidium iodide staining, revealing that phloroglucinol markedly reduced high glucose‐induced apoptosis. These results demonstrated that phloroglucinol could be useful as a potential therapeutic agent for the protection of pancreatic β‐cells against glucose‐induced apoptosis. Copyright © 2015 John Wiley & Sons, Ltd.     

1,2,3,6‐tetra‐O‐galloyl‐β‐D‐allopyranose gallotannin isolated,from Euphorbia jolkini,attenuates LPS‐induced nitric oxide production in macrophages

Nitric oxide (NO) is a pleiotropic regulator, critical to numerous biological processes, including vasodilatation and macrophage‐mediated immunity. Macrophages express inducible NO synthase (iNOS) and produce NO after lipopolysaccharide (LPS) stimulation. Gallotannins are water‐soluble polyphenols with wide‐ranging biological activities. Various chemical structures of gallotannins occurring in medicinal and food plants that are used worldwide showed several remarkable biological and pharmacological activities. In the present study, we examined the inhibitory effects of gallotannin 1,2,3,6‐tetra‐O‐galloyl‐β‐D‐allopyranose (GT24) isolated from Euphorbia jolkini on the LPS‐induced NO production and underlying mechanisms of action. GT24 dose‐dependently decreased LPS‐induced NO production and iNOS expression in J774A.1 macrophages. In addition, GT24 inhibited LPS‐induced activation of nuclear factor (NF)‐κB as indicated by inhibition of degradation of I‐κBα, nuclear translocation of NF‐κB, and NF‐κB dependent gene reporter assay. Our results suggest that GT24 possesses an inhibitory effect on the LPS‐induced inflammatory reaction. Copyright © 2010 John Wiley & Sons, Ltd.     

A Comparative Study on the Inhibitory Effects of Different Parts and Chemical Constituents of Pomegranate on α‐Amylase and α‐Glucosidase

Pomegranate has been documented for the management of diabetes in Unani and Chinese medicine. This study compared the effects of the extracts of different pomegranate parts, including juice, peels, seeds and flowers, on carbohydrate digestive enzymes (α‐amylase and α‐glucosidase) in vitro. The methanolic flower extract inhibited α‐amylase and α‐glucosidase, while the methanolic peel extract inhibited α‐glucosidase selectively. The most active flower extract was subjected to water‐ethyl acetate partition. The ethyl acetate fraction was more potent than the water fraction in inhibiting both enzymes. Gallic acid and ellagic acid also showed selective inhibition against α‐glucosidase, and their presence in the ethyl acetate fraction was confirmed by HPLC‐DAD and HPLC‐HESI‐MS. Our findings suggest that the inhibition of carbohydrate digestive enzymes and their phenolic content may contribute to the anti‐hyperglycaemic effects of pomegranate flower and peel, and support their claims in diabetes. Copyright © 2012 John Wiley & Sons, Ltd.     

冬凌草甲素抑制血管生成活性及作用机制

目的:研究冬凌草甲素抑制血管生成的活性及其作用机制。方法:采用体外和体内两种模式,研究冬凌草甲素在血管生成方面的抑制效应。通过体外培养心脏微血管内皮细胞,噻唑蓝(MTT)比色法检测细胞的活力,酶联免疫吸附(ELISA)法测定血管内皮生长因子(VEGF)的含量;体内选择斑马鱼(Fli1-GFP)生物模式,观察冬凌草甲素对胚胎期血管生成和成鱼期血管损伤后再生的影响,并采用相对荧光定量PCR法检测VEGF通路上主要相关基因VEGFA,血管内皮细胞生长因子受体(VEGFR)2,VEGFR3的表达量。结果:冬凌草甲素具有抑制血管生成活性的作用,可抑制体外内皮细胞活力,其半抑制浓度(IC50)8.04 mg·L~(-1),可使细胞血清中VEGF的表达量明显下降;可抑制斑马鱼胚胎期体节间血管生成,且抑制成鱼期血管损伤后再生,可能通过降低体内VEGFA,VEGFR2,VEGFR3基因的表达量从而抑制血管生成。结论:冬凌草甲素可有效抑制血管生成,为其抗肿瘤血管生成治疗提供了重要的科学依据。     

Toll‐like receptor 4‐mediated immunoregulation by the aqueous extract of Mori Fructus

The aqueous extract of Mori Fructus (MF) exerts a change of phenotype and a cytoprotective effect in macrophages. The present study was carried out to investigate the immunomodulating activity of MF on the expression of nitric oxide (NO), tumor necrosis factor alpha (TNF‐α), co‐stimulatory molecules and also interferon‐gamma (IFN‐γ) in macrophages and splenocytes. Toll‐like receptor 4 (TLR4) is a promising molecular target for immune‐modulating drugs. It was hypothesized that one possible upstream signaling pathway leading to immunoregulation of MF may be mediated by TLRs. Multiple signaling molecules (NF‐κB, ERK1/2, p38 and JNK) of the TLR4 signaling pathway were also detected. It was found that MF increased NO production and TNF‐α secretion in RAW 264.7 and peritoneal macrophages, co‐stimulatory molecules expression in peritoneal macrophages and IFN‐γ expression in splenocytes. Further studies indicated that MF could significantly induce the phosphorylation of signal molecules of MAPKs and the degradation of IκBα which finally led to the activation and nuclear translocation of nuclear factor‐κB (NF‐κB) for the target gene expression. All those notions disclosed that the aqueous extract MF is a new TLR4 activator, which induces a Th1 immune response as a consequence of induction of cytokines secretion, especially TNF‐α and IFN‐γ. Copyright © 2009 John Wiley & Sons, Ltd.