The Androgenic Alopecia Protective Effects of Forsythiaside‐A and the Molecular Regulation in a Mouse Model

This study examined the inhibitory effect of forsythiaside‐A, a natural substance derived from Forsythia suspensa (F. suspensa), on entry into catagen induced by dihydrotestosterone (DHT) in an androgenic alopecia mouse model. In vitro experiment comparing finasteride with forsythiaside‐A showed that forsythiaside‐A treatment resulted in a 30% greater inhibition of DHT‐induced apoptosis in human hair dermal papilla cell (HHDPCs) and human keratinocytes (HaCaTs). In vivo experiment showed that mouse hair density and thickness were increased by 50% and 30%, respectively, in the forsythiaside‐A‐treated group when compared to a DHT group. Tissue histological results revealed that the forsythiaside‐A‐treated group had an increase in size and shape of the hair follicles and a 1.5 times increase in the follicle anagen/telogen ratio when compared to the finasteride group. Western blot examination of TGF‐β2 expression related to apoptosis signaling in mouse skin verified that forsythiaside‐A reduced the expression of TGF‐β2 by 75% and suppressed apoptosis by reducing the expression of caspase‐9 by 40%, and caspase‐3 by 53%, which play an roles up‐regulator in the apoptosis signal. The forsythiaside‐A group also showed a 60% increase in the Bcl‐2/Bax ratio, which is a factor related to mitochondrial apoptosis. Our results indicated that forsythiaside‐A prevents apoptosis by similar mechanism with finasteride, but forsythiaside‐A is more effective than finasteride. In summary, forsythiaside‐A controlled the apoptosis of hair cells and retarded the entry into the catagen phase and therefore represents a natural product with much potential for use as a treatment for androgenic alopecia. Copyright © 2015 John Wiley & Sons, Ltd.     

Topical application of Polygonum multiflorum extract induces hair growth of resting hair follicles through upregulating Shh and β-catenin expression in C57BL/6 mice

Ethnopharmacological relevance

Polygonum multiflorum has traditionally been used for treating patients suffering from baldness and hair loss in East Asia.

Aim of the study

The present study sought to investigate the hair growth promoting activities of Polygonum multiflorum and its mechanism of action.

Materials and methods

The Polygonum multiflorum extract was topically applied to the shaved dorsal skin of telogenic C57BL6/N mice. To determine the effect of Polygonum multiflorum extract in telogen to anagen transition, the expression of β-catenin and Sonic hedgehog (Shh) was determined by immunohistochemistry analysis.

Results

Polygonum multiflorum extract promoted hair growth by inducing anagen phase in telogenic C57BL6/N mice. In Polygonum multiflorum extract treated group, we observed increase in the number and the size of hair follicles that are considered as evidence for anagen phase induction. Immunohistochemical analysis revealed that earlier induction of β-catenin and Shh were observed in Polygonum multiflorum extract treated group compared to that in control group.

Conclusion

These results suggest that Polygonum multiflorum extract promotes hair growth by inducing anagen phase in resting hair follicles.     

The Effect of the Ginkgo biloba Extract in the Expression of Bax,Bcl‐2 and Bone Mineral Content of Wistar Rats with Glucocorticoid‐Induced Osteoporosis

Objective: Evaluate the effects of the extract of Ginkgo biloba (EGb) in the glucocorticoid‐induced‐osteoporosis through the Bax and Bcl‐2 expressions by osteoblast cells, the x‐ray and bone density of the tibia. Method: Rats were divided into five groups: osteoporosis; EGb1 (28 mg/kg); EGb2 (56 mg/kg); alendronate (0.2 mg/animal) and control. The treatments were conducted for 20 (n = 30) and 30 days (n = 30). The Bax and Bcl‐2 expressions were evaluated in osteoblasts of the mandibular alveolar bone. The tibias were radiographed to evaluate the X‐ray and bone density. The control group was compared with the osteoporosis' (Student's t‐test/Mann‐Whitney). The other groups were analyzed by analysis of variance test followed by Dunnett/Dunnett T3 (p < 0.05). Results: When compared the osteoporosis to the control group (p <0.05): Bax and x‐ray density increased; Bcl‐2 and the bone density reduced. When compared with the osteoporosis group (p < 0.05), alendronate (30 days), EGb1 and EGb2 (20/30 days) increased the Bcl‐2 expression; EGb2 and alendronate (20 days) EGb1 and EGb2 (30 days) reduced the Bax expression; and EGb1 and EGb2 (20/30 days) reduced the X‐ray density. Conclusions: The EGb improved the Bcl‐2 and reduced the Bax expression by osteoblasts in the mandibular alveolar bone and recovered the mineral content in the tibia of rats with glucocorticoid‐induced‐osteoporosis. Copyright © 2012 John Wiley & Sons, Ltd.     

Ginsenosides Rb1 and Rd Regulate Proliferation of Mature Keratinocytes Through Induction of p63 Expression in Hair Follicles

Ginsenosides Rb₁ and Rd are the two main types of ginsenosides in Panax ginseng and have been used as an additive to against alopecia. However, the mechanisms involved are largely unknown. To determine how ginsenosides prevent hair loss, we topically applied protopanaxadiol‐type ginsenosides Rb₁ and Rd over the shaved skin of B57CL/6 mice, and monitored and assessed them for 35 days. We then investigated the effects of ginsenosides on cell genesis in different phases of adult hair follicles (HFs), using 5‐bromo‐2′‐deoxyuridine as a marker for dividing cells. Moreover, p63, a specific marker and a major regulator of keratinocyte progenitor cells of the multi‐layered epithelia, was detected in epidermis. Results indicated that treatment with ginsenosides Rb₁ and Rd increased cell proliferation in both anagen and telogen of HFs. However, it had no significant effect on the survival of cells in the bulge and upper follicle region. Investigation of p63 demonstrated that up‐regulation of p63 expression in the matrix and outer root sheath might be one of the mechanisms by which ginsenosides Rb₁ and Rd promote cell proliferation in HFs. Our study reveals a novel mechanism by which ginsenoside promotes hair growth through p63 induction in follicular keratinocytes and indicates that ginsenosides Rb₁ and Rd might be developed as a therapeutic agent for the prevention of hair loss. Copyright © 2012 John Wiley & Sons, Ltd.     

Hair Growth‐Promoting Effect of Carthamus tinctorius Floret Extract

The florets of Carthamus tinctorius L. have traditionally been used for hair growth promotion. This study aimed to examine the potential of hydroxysafflor yellow A‐rich C. tinctorius extract (CTE) on hair growth both in vitro and in vivo. The effect of CTE on cell proliferation and hair growth‐associated gene expression in dermal papilla cells and keratinocytes (HaCaT) was determined. In addition, hair follicles from mouse neonates were isolated and cultured in media supplemented with CTE. Moreover, CTE was applied topically on the hair‐shaved skin of female C57BL/6 mice, and the histological profile of the skin was investigated. C. tinctorius floret ethanolic extract promoted the proliferation of both dermal papilla cells and HaCaT and significantly stimulated hair growth‐promoting genes, including vascular endothelial growth factor and keratinocyte growth factor. In contrast, CTE suppressed the expression of transforming growth factor‐β1 that is the hair loss‐related gene. Furthermore, CTE treatment resulted in a significant increase in the length of cultured hair follicles and stimulated the growth of hair with local effects in mice. The results provided the preclinical data to support the potential use of CTE as a hair growth‐promoting agent. Copyright © 2013 John Wiley & Sons, Ltd.     

Flavonoids of Korean Citrus aurantium L. Induce Apoptosis via Intrinsic Pathway in Human Hepatoblastoma HepG2 Cells

Korean Citrus aurantium L. has long been used as a medicinal herb for its anti‐inflammatory, antioxidant, and anticancer properties. The present study investigates the anticancer role of flavonoids extracted from C. aurantium on human hepatoblastoma cell, HepG2. The Citrus flavonoids inhibit the proliferation of HepG2 cells in a dose‐dependent manner. This result was consistent with the in vivo xenograft results. Apoptosis was detected by cell morphology, cell cycle analysis, and immunoblot. Flavonoids decreased the level of pAkt and other downstream targets of phosphoinositide‐3‐kinase/Akt pathway – P‐4EBP1 and P‐p70S6K. The expressions of cleaved caspase 3, Bax, and Bak were increased, while those of Bcl‐2 and Bcl‐xL were decreased with an increase in the expression of Bax/Bcl‐xL ratio in treated cells. Loss of mitochondrial membrane potential was also observed in flavonoid‐treated HepG2 cells. It was also observed that the P‐p38 protein level was increased both dose and time dependently in flavonoid‐treated cells. Collectively, these results suggest that flavonoid extracted from Citrus inhibits HepG2 cell proliferation by inducing apoptosis via an intrinsic pathway. These findings suggest that flavonoids extracted from C. aurantium L. are potential chemotherapeutic agents against liver cancer. Copyright © 2015 John Wiley & Sons, Ltd.     

Fructus panax ginseng extract promotes hair regeneration in C57BL/6 mice

Ethnopharmacological relevance

Radix panax ginseng (Panax ginseng C.A. Meyer, Araliaceae, RPG) has been documented to possess hair growth activity and widely used to treat alopecia, while no report has been issued to date on the effect of Fructus panax ginseng (FPG) on hair regeneration.

Materials and methods

To investigate the effects of FPG extract on the proliferation of human hair dermal papilla cells (DPCs) and on the promotion of hair regeneration in C57BL6 mice, cell proliferation was evaluated in cultured DPC by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) and measured the expressions of Bcl-2 and Bax by immunoblot assay. We also compared the effects of topical FPG extract (1 and 10 mg/ml, 100 μl/d) with the effects of minoxidil as a positive control (5%, 100 μl/d) or vehicle control (30% ethanol) on the depilation-induced hair cycling in 7 week-old-C57BL/6 mice.

Results

FPG extract significantly increased the proliferation of DPCs in dose and time dependent manners (P < 0.05, P < 0.01 and P < 0.001). FPG extract also enhanced Bcl-2 expression and decreased Bax expression compared with control (P < 0.01). Moreover, significant elongations of anagen phase during hair cycle after application of FPG were evaluated by photographical and histological observations.

Conclusions

FPG extract improves the cell proliferation of human DPCs through anti apoptotic activation. Topical administration of FPG extract might have hair regeneration activity for the treatment of hair loss.     

人参提取物对C57小鼠生发作用的影响

目的: 研究人参提取物对C57BL/6J小鼠毛发生长促进作用及其机制。 方法: 实验分为阳性对照组(章光101)、空白对照组、人参提取物高、中、低(2, 1, 0.5 g·mL^-1)剂量组。小鼠脱毛,通过观察脱毛区肤色的变化和组织学检查,研究人参提取物对小鼠毛发生长周期的影响;采用红外测温仪观察对小鼠皮肤温度的影响;采用浊度法和底物发色法,观察其抗血小板聚集的作用,初步探讨其促进毛发生长的机制。 结果: 人参各剂量组均能诱导C57BL/6J小鼠毛发生长,使其从休止期进入生长期,但对毛发的最终长度无影响,且呈现一定的剂量依赖作用(P<0.01),高剂量组与阳性组无显著性差异;组织学检查显示人参提取物可以刺激毛囊生长,产生黑色素;给药20,60 min后,人参各剂量组均能显著提高小鼠的体温(P<0.01),120 min后,恢复到给药前水平;人参高、中、低剂量组对血小板活化因子(PAF)诱导的大鼠5 min时血小板聚集率分别为49.41%,27.80%,18.39%(均P<0.01),且呈现一定的剂量依赖作用,与阳性对照组则无显著性差异;其抗血小板聚集的作用与抗凝血酶有关, 人参高、中、低剂量组的凝血酶抑制率分别为(20.24±2.72)%,(15.42±1.03)%,(8.61±0.80)%(P<0.01)。 结论: 人参提取物具有明显的促进毛发生长的作用,其机制与扩张皮肤毛细血管、改善局部微循环有关。     

Inhibitory Effects of Nobiletin on Hepatocellular Carcinoma In Vitro and In Vivo

Nobiletin (5, 6, 7, 8, 3′ 4′‐hexamethoxyflavone) is a major anticancer component in juice from zhishi (Rutaceae). This study aimed to investigate the inhibitory effect of Nobiletin on hepatic cancer cells both in vitro and in vivo. The 3‐(4, 5‐dimethylthiazol‐2‐yl)‐2, 5‐diphenyltetrazolium bromide (MTT), growth curve, and clonogenic assay showed that nobiletin inhibited the proliferation of SMMC‐7721 cells in vitro. Hoechst staining observed the characteristics of cell apoptosis in nobiletin‐treated cells, and the apoptotic rates of treated groups were increased in a dose‐dependent manner. Flow cytometric analysis demonstrated that nobiletin could block the cell cycle arrested at G2 phase. Cell cycle analysis was performed using flow cytometry. Results showed that cell cycle phase distribution analysis showed G2 arrest. It was found that nobiletin downregulated the expressions of Bcl‐2 and COX‐2 and up‐regulated the expressions of Bax and caspase‐3 in SMMC‐7721 cells by western blotting. The experiment in vivo demonstrated that nobiletin significantly inhibited the growth of H22 transplantable tumor, downregulated the expressions of COX‐2, up‐regulated the expressions of Bax and caspase‐3 detected by immunohistochemistry and western blotting, and the ratios of Bcl‐2/Bax were decreased. Our results suggest that nobiletin has significant inhibitory effects on hepatocellular carcinoma both in vitro and in vivo. Copyright © 2013 John Wiley & Sons, Ltd.     

他克莫司对小鼠触须毛囊体外培养生物学特性的研究

 目的 观察不同浓度他克莫司(FK506)对小鼠触须游离毛囊器官培养的生物学特性影响。方法 用离体器官培养的方法在倒置显微镜底下每日测量不同浓度FK506作用下毛囊生长长度、观察毛囊的形态学、记录生长天数,以及液相闪烁仪测量同位素³H-TdR的掺入率。结果0.003-0.3 mg·L^-1浓度的FK506能促进毛囊的生长和DNA的合成,推迟生长期毛囊向退行期转变,延长毛囊的生长时间。当FK506浓度大于1 mg·L^-1时即明显抑制毛囊在体外的生长。结论 FK506在体外能直接促进毛囊的生长,延长毛囊的生长时间。     

Oral administration of Aloe vera and honey reduces walker tumour growth by decreasing cell proliferation and increasing apoptosis in tumour tissue

Cancer is diagnosed in approximately 11 million people and is responsible for almost 8 million deaths worldwide every year. Research in cancer control has shown the importance of co‐adjuvant therapies. Aloe vera may reduce tumour mass and metastasis rates, while honey may inhibit tumour growth. This study verified the influence of Aloe vera and honey on tumour growth and in the apoptosis process by assessing tumour size, the cell proliferation rate (Ki67‐LI) and Bax/Bcl‐2 expression at 7, 14 and 20 days after Walker 256 carcinoma implant in Wistar rats distributed into two groups: the WA group – tumour‐bearing rats that received a gavage with a 670 µL/kg dose of Aloe vera and honey solution daily, and the CW group – tumour‐bearing rats which received only a 0.9% NaCl solution. The effect of Aloe vera and honey against tumour growth was observed through a decrease in relative weight (%) and Ki67‐LI in tumours from the WA group compared with those from the CW group. The Bax/Bcl‐2 ratio increased in tumours from the WA group at all tested timepoints. These data suggest Aloe vera and honey can modulate tumour growth by reducing cell proliferation and increasing apoptosis susceptibility. Copyright © 2010 John Wiley & Sons, Ltd.     

Comparative investigations on the protective effects of rhodioside, ciwujianoside-B and astragaloside IV on radiation injuries of the hematopoietic system in mice

The aim of this study was to investigate the protective effects of three glycosides (rhodioside, ciwujianoside‐B and astragaloside IV) on the hematopoietic system in the mice exposed to γ‐rays, and to examine the possible mechanisms involved. Mice were pretreated with the glycosides (40 mg/kg, i.g.) daily for 7 days prior to radiation. The survival of mice pretreated with three glycosides after total body irradiation (6.0 Gy) was examined. Peripheral blood leucocytes and endogenous spleen colony counts, colony‐forming unit‐granulocyte macrophage assay, analysis of DNA content and apoptosis rate determination were performed to evaluate the effects of the three glycosides on hematogenesis. The fragmentation of double‐stranded DNA in lymphocytes was detected by the comet assay. The changes in cell cycle were analysed by flow cytometry. Furthermore, the expression levels of Bcl‐2, Bax and nuclear factor‐kappa B (NF‐κB) were measured by western blot and the electrophoretic mobility shift assay. The results showed that pretreatment with all of the glycosides improved survival time and increased the number of leucocytes, spleen colonies and granulocyte‐macrophage colonies in mice exposed to 6.0 Gy γ‐radiation. Rhodioside showed more protective efficacy than both ciwujianoside‐B and astragaloside IV. All three glycosides significantly increased the proliferation abilities of bone marrow cells, and decreased the ratio of cells in G₀/G₁ phase. Further analysis showed that these three glycosides were able to decrease DNA damage and the increment in the Bax/Bcl‐2 ratio induced by radiation. In summary, the three glycosides showed radioprotective effects on the hematopoietic system in mice, which was associated with changes in the cell cycle, a reduction in DNA damage, and down‐regulation of the ratio of Bax/Bcl‐2 in bone marrow cells exposed to radiation. Copyright © 2010 John Wiley & Sons, Ltd.     

Effect of plum-blossom needle tapping with different stimulation intensities on hair regrowth in hair removal mice

Objective

To determine the role of intensity of plum-blossom needle tapping in treating alopecia areata.

Methods

The BALB/c mice were randomized into a normal group, a control group, a roller-needle (RN) group, a mild plum-blossom needle (MP) group, and a heavy plum-blossom needle (HP) group. An area of hair was removed by external application of 8% sodium sulfide on BALB/c mice. The hair regrowth, hair follicle changes, and local inflammatory factor changes after cutaneous acupuncture were observed.

Results

After treated with sodium sulfide, the hair was completely removed, the local hair follicles reached the catagen phase, and the expressions of interleukin (IL)-1α, IL-1β, IL-2, tumor necrosis factor (TNF)-α, interferon (IFN)-γ, and IL-17 were increased. Mice intervened by RN achieved the same hair growth rating as the controls but with thicker hair shafts; mice in the MP group had incomplete and uneven hair growth but thicker hair shafts; mice in the HP group didn’t show hair growth. Pathological analysis revealed significant inflammatory infiltration into the local follicle bulbs and increased catagen-phase follicles in the control group, while RN and MP groups showed significantly increased anagen-phase follicles, coarser individual hairs, and obvious hair shafts. Meanwhile, most of the hair follicles in the HP group were in telogen phase and showed obvious surrounding inflammatory infiltration. RN, MP, and HP significantly down-regulated the increased IL-2, IL-1α, IL-1β, TNF-α, and IFN-γ levels (P<0.05), but didn’t notably affect the increased CD34 expression (P>0.05).

Conclusion

Cutaneous acupuncture with heavy stimulation intensity can inhibit hair growth in hair removal mice, while RN, with the lightest stimulation intensity, is unlikely to affect hair growth but may make hair shafts thicker and follicles larger.

     

Corosolic Acid Triggers Mitochondria and Caspase‐dependent Apoptotic Cell Death in Osteosarcoma MG‐63 Cells

The response of osteosarcoma MG‐63 cells to corosolic acid treatment has been investigated. The results showed that corosolic acid significantly inhibited cell viability in both a dose and a time dependent manner. It was found that corosolic acid increased the Bax/Bcl‐2 ratio by up‐regulating Bax expression, disrupted mitochondrial membrane potential and triggered the release of cytochrome c from mitochondria into the cytoplasm. Corosolic acid treatment triggered the activation of caspase‐8, 9 and 3. The apoptosis was obviously inhibited by pretreatment with a general caspase inhibitor, z‐VAD‐FMK. Moreover, pretreatment of CsA, a cyclophilin D ligand that inhibits mitochondria potential uncoupling, prevented the activation of caspase‐9 and caspase‐3, but not caspase‐8, and the apoptosis of MG‐63 cells, triggered by corosolic acid. All these results indicated that corosolic acid‐induced apoptosis was associated with the activation of caspases via a mitochondrial pathway. Copyright © 2011 John Wiley & Sons, Ltd.     

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.     

Emodin Regulates Apoptotic Pathway in Human Liver Cancer Cells

Emodin, a natural anthraquinone, has been reported to possess antiproliferative effects in many cancer cell lines. However, anticancer mechanism against human liver cancer remains unclear. In this study, we observed that emodin induced apoptosis in HepG2 cells and caused a significant accumulation of cells in the G1 phase. Western blot data showed that emodin treatment caused the increasing of release of cytochrome c into cytosol from mitochondria and the activation of caspase‐8 and caspase‐9, which suggest that the intrinsic and extrinsic pathways could be involved. Emodin treatment also resulted in a dose‐dependent accumulation of intracellular reactive oxygen species. Furthermore, emodin increased the protein level of p53 and decreased the protein level of NF‐κB/p65 in HepG2 cells, which indicated these two regulators might play a role in emodin‐induced apoptosis. Computational modeling showed that emodin could directly bind to the BH3 domain of Bcl‐2 through forming one hydrogen bond with Ala146 residue in Bcl‐2. From these examinations, emodin not only significantly downregulated expression of Bcl‐2 but also inhibited the heterodimerization of Bcl‐2 with Bax because of strong interaction between emodin and Bcl‐2. These suggest that emodin induces apoptosis in liver cancer cell line through a multifaceted complex cascade of events. Copyright © 2012 John Wiley & Sons, Ltd.     

Antiproliferative Activity of Phenylpropanoids Isolated from Lagotis brevituba Maxim

The aim of the present study was to evaluate the antiproliferative effect of phenylpropanoids isolated from the n‐BuOH‐soluble fraction of an ethanolic extract of Lagotis brevituba Maxim. The phenylpropanoids were identified as echinacoside, lagotioside, glucopyranosyl(1–6)martynoside, plantamoside, and verbascoside. Three of the compounds, lagotioside, glucopyranosyl(1–6)martynoside, and plantamoside, were isolated from L. brevituba for the first time. The antiproliferative activity of the isolates was evaluated in human gastric carcinoma (MGC‐803), human colorectal carcinoma (HCT116), human hepatocellar carcinoma (HepG2), and human lung cancer (HCT116) cells using an 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide assay. Plantamoside showed promising activity against MGC‐803 cells, with a half maximal inhibitory concentration value of 37.09 μM. The mechanism of the pro‐apoptosis effect of plantamoside was then evaluated in MGC‐803 cells. Changes in cell morphology, including disorganization of the architecture of actin microfilaments and formation of apoptotic bodies, together with cell cycle arrest in G2/M phases, were observed after treatment of plantamoside. The antiproliferative and pro‐apoptotic effects were associated with a decrease in the ratio of Bcl‐2/Bax and reduced mitochondrial membrane potential, which was accompanied by the release of reactive oxygen species and Ca²⁺ into the cytoplasm. Taken together, the results indicated that plantamoside promotes apoptosis via a mitochondria‐dependent mechanism. Copyright © 2017 John Wiley & Sons, Ltd.     

Ocimum sanctum induces apoptosis in A549 lung cancer cells and suppresses the in vivo growth of lewis lung carcinoma cells

Although Ocimum sanctum has been used extensively for its medicinal values in India and China, its antitumor activity against human nonsmall cell lung carcinoma (NSCLC) A549 cells has not been investigated until now. Therefore, the antitumor mechanism of ethanol extracts of Ocimum sanctum (EEOS) was elucidated in A549 cells in vitro and the Lewis lung carcinoma (LLC) animal model. EEOS exerted cytotoxicity against A549 cells, increased the sub‐G1 population and exhibited apoptotic bodies in A549 cells. Furthermore, EEOS cleaved poly(ADP‐ribose)polymerase (PARP), released cytochrome C into cytosol and simultaneously activated caspase‐9 and ‐3 proteins. Also, EEOS increased the ratio of proapoptotic protein Bax/antiapoptotic protein Bcl‐2 and inhibited the phosphorylation of Akt and extracellular signal regulated kinase (ERK) in A549 cancer cells. In addition, it was found that EEOS can suppress the growth of LLC inoculated onto C57BL/6 mice in a dose‐dependent manner. Overall, these results demonstrate that EEOS induces apoptosis in A549 cells via a mitochondria caspase dependent pathway and inhibits the in vivo growth of LLC, suggesting that EEOS can be applied to lung carcinoma as a chemopreventive candidate. Copyright © 2009 John Wiley & Sons, Ltd.     

Curcumin Inhibits Proliferation and Induces Apoptosis of Human Colorectal Cancer Cells by Activating the Mitochondria Apoptotic Pathway

Curcumin, a natural plant extract from Curcuma longa, is known for its anti‐carcinogenic and chemopreventive effects on a variety of experimental cancer models. In this study, we evaluated the effects of curcumin and elucidated its mechanism in human colorectal carcinoma cells. Cell viability assay showed that curcumin significantly inhibited the growth of LoVo cells. Curcumin treatment induced the apoptosis accompanied by ultra‐structural changes and release of lactate dehydrogenase in a dose‐dependent manner. Moreover, treatment with 0–30 µg/mL curcumin decreased the mitochondrial membrane potential and activated the caspase‐3 and caspase‐9 in a dose‐ and time‐dependent manner. Nuclear and annexin V/PI staining showed that curcumin induced the apoptosis of LoVo cells. FACS analysis revealed that curcumin could induce the cell cycle arrest of LoVo cells at the S phase. Furthermore, western blotting analysis indicated that curcumin induced the release of cytochrome c, a significant increase of Bax and p53 and a marked reduction of Bcl‐2 and survivin in LoVo cells. Taken together, our results suggested that curcumin inhibited the growth of LoVo cells by inducing apoptosis through a mitochondria‐mediated pathway. Copyright © 2012 John Wiley & Sons, Ltd.