Thai traditional medicines reduce CD147 levels in lung cells: Potential therapeutic candidates for cancers, inflammations, and COVID-19.

ETHNOPHARMACOLOGICAL RELEVANCE: The cluster of differentiation 147 (CD147) is identified as the signaling protein relevant importantly in various cancers, inflammations, and coronavirus disease 2019 (COVID-19) via interacting with extracellular cyclophilin A (CypA). The reduction of CD147 levels inhibits the progression of CD147-associated diseases. Thai traditional medicines (TTMs): Keaw-hom (KH), Um-ma-ruek-ka-wa-tee (UM), Chan-ta-lee-la (CT), and Ha-rak (HR) have been used as anti-pyretic and anti-respiratory syndromes caused from various conditions including cancers, inflammations, and infections. Thus, these medicines would play a crucial role in the reduction of CD147 levels. AIM OF THE STUDY: This article aimed to investigate the effects of KH, UM, CT, and HR for reducing the CD147 levels through in vitro study. Additionally, in silico study was employed to screen the active compounds reflexing the reduction of CD147 levels. MATERIALS AND METHODS: The immunofluorescent technique was used to evaluate the reduction of CD147 levels in human lung epithelial cells (BEAS-2B) stimulated with CypA for eight extracts of KH, UM, CT, and HR obtained from water decoction (D) and 70% ethanol maceration (M) including, KHD, UMD, CTD, HRD, KHM, UMM, CTM, and HRM. RESULTS: UM extracts showed the most efficiency for reduction of CD147 levels in the cytoplasm and perinuclear of BEAS-2B cells stimulated with CypA. Phenolic compounds composing polyphenols, polyphenol sugars, and flavonoids were identified as the major chemical components of UMD and UMM. Further, molecular docking calculations identified polyphenol sugars as CypA inhibitors. CONCLUSIONS: UMD and UMM are potential for reduction of CD147 levels which provide a useful information for further development of UM as potential therapeutic candidates for CD147-associated diseases such as cancers, inflammations, and COVID-19.

Formulation and In Vitro Evaluation of Mucoadhesive Sustained Release Gels of Phytoestrogen Diarylheptanoids from Curcuma comosa for Vaginal Delivery.

Diarylheptanoids (DAs) characterized by a 1,7-diphenylheptane structural skeleton are considered a novel class of phytoestrogens. The DAs available in Curcuma comosa Roxb. (C. comosa) extract demonstrated significant estrogenic activities both in vitro and in vivo. This study aimed to develop and comprehensively evaluate a mucoadhesive vaginal gel for the sustained release of DAs. Different mucoadhesive polymers as gelling agents were investigated. C. comosa ethanolic crude extract was used as a source of DAs. All C. comosa gels were light brown homogeneous with pH within 4.4-4.6. Their flow behaviors were pseudoplastic with a flow behavior index of 0.18-0.38. The viscosity at a low shear rate varied from 6.2 to 335.4 Pa·s. Their mechanical and extrudability properties were associated well with rheological properties. Polycarbophil (PCP):hydroxypropyl methylcellulose (HPMC) blends had a higher mucoadhesiveness to porcine vaginal mucosa than those of PCP-based or HPMC-based gels. All C. comosa gels exhibited a sustained, zero-order DA release pattern over 72 h. Korsmeyer and Peppas equation fitting indicated a non-Fickian, case II transport release mechanism. C. comosa gels had good physical and chemical stability under low-temperature storage for up to 12 months. PCP:HPMC-based mucoadhesive gels could be a proper delivery system for vaginal administration of DAs.

Supplementation of Kaempferia parviflora Extract Enhances Physical Fitness and Modulates Parameters of Heart Rate Variability in Adolescent Student-Athletes: A Randomized, Double-Blind, Placebo-Controlled Clinical Study.

This randomized double-blind controlled study aimed to investigate the effects of a standardized Kaempferia parviflora (KP) extract on the physical fitness and heart rate variability (HRV) parameters in adolescent sport school students. 194 male students were recruited and randomized into two groups (n = 97), matched by age and sports. The KP-treated group received KP extract capsules at a dose of 360 mg/day and the control group received placebo capsules, continuously for 12 weeks. Physical fitness performance and HRV parameters were monitored with blood biochemical analysis for product safety. KP extract significantly increased the right-hand grip strength, the back-leg strength and maximal oxygen consumption (VO2 max) and decreased the time used for 50-meter sprint test without changing the sit-and-reach test and the 40 yard technical test. For HRV parameters, KP extract significantly increased standard deviation of normal to normal intervals (SDNN), square root of the mean of square of successive normal to normal interval differences (RMSSD) and high frequency (HF) norm, without changing low frequency (LF) norm and LF/HF ratio. The increase in stress resistance and decrease in stress index were found in the KP-treated group, without changing the autonomic nervous system (ANS) activity and balance. Blood biochemical analysis showed normal values of all participants. This data indicates the safety and positive effects of KP on muscle strength, endurance and speed, but not on the flexibility and agility. The modulatory effects of KP extract on HRV parameters suggest its anti-stress effects and would encourage the application in a sport training and exercise.

Bioactivities of Lentinus polychrous and Ganoderma lucidum (Agaricomycetes) Polysaccharide Extracts.

This study aimed to compare the bioactivities of Lentinus polychrous polysaccharide extracts with Ganoderma lucidum. Three hot water-extracted polysaccharide fractions of L. polychrous (LPE), including immature fruiting bodies (im-LPE), mature fruiting bodies (ma-LPE), and mycelium (my-LPE), were analyzed for their phytochemical contents and bioactivities (e.g., anticancer, antiviral, anti-inflammatory, and immunomodulatory effects) compared with G. lucidum extract (GLE). Although GLE had the highest total phenolic and protein contents and lower carbohydrate content than LPE, im-LPE showed strong inhibition on HepG2 cell proliferation as did GLE. GLE and LPE showed antiviral effects on herpes simplex virus-1 and the relative antiproliferative activity was from im-LPE > my-LPE > ma-LPE. However, im-LPE showed the best protective activity on the viral attachment step with some virucidal effects. Moreover, all LPE showed selective and stronger inhibitory effects on LPS-induced RAW264.7 macrophages than those of GLE on genetic expression. Considering the 50% inhibitory concentration values, my-LPE possessed the strongest inhibitory activity on the expression of cyclooxygenase-2 and inducible nitric oxide synthase. However, ma-LPE had the strongest inhibitory effects on interleukin-1ß and tumor necrosis factor-α gene expression. The extracts increased splenocyte proliferation under mycohaemagglutinin induction with a relative order of my-LPE > GLE > ma-LPE = im-LPE. In conclusion, LPE showed positive effects and stronger activity than G. lucidum. L. polychrous may have potential for use as an ingredient in functional foods.

Curcuma comosa loaded transfersomal gel for transdermal application: formulation, in vitro and in vivo evaluation.

RATIONAL: Diarylheptanoids, extracted from the Curcuma comosa (CC) rhizome, have been reported to exhibit estrogenic activity. However, oral administration of the extract showed a short half-life. OBJECTIVES: This study aimed to formulate and to investigate the potential of transfersomal gels for the transport of phytoestrogenic diarylheptanoids across the skin into the blood circulation. MATERIALS AND METHODS: The transfersomes were developed and optimized for their compositions including sources of phospholipid (egg yolk and soybean), types of edge activators (polysorbate 80, sorbitan oleate 80, and sodium cholate), and concentrations of CC extract (10-60 mg). The optimal formulation was further incorporated into Carbopol® Ultrez 21 gel and evaluated for in vitro release, permeation, and in vivo absorption. RESULTS: The optimal transfersomes containing 10% of polysorbate 80 were selected due to high drug entrapment efficiency and a small diameter. The release kinetic of transfersomal gels followed a zero model. The maximum permeation flux through porcine ear skin was 1.38 ± 0.25 µg/cm2/h for (4E, 6E)-1, 7-diphenylhepta-4, 6-dien-3-ol, and 0.40 ± 0.11 µg/cm2/h for (6E)-1, 7-diphenylhept-6-en-3-ol. Results of the in vivo pharmacokinetics study in rats showed that transfersomal gel provided a maximum concentration of 219.71 ± 4.05 ng/ml and prolonged plasma concentration of diarylheptanoids for over 12 h. There was no significant variation found in the physical characteristics including viscosity, pH, and size after six months of storage at room temperature (30 ± 1 °C) and high temperature (40 ± 1 °C). CONCLUSIONS: The obtained data suggested that the developed transfersomal gel of CC extract should be beneficial for improving the delivery of phytoestrogenic diarylheptanoids.

Antitumor Efficacy of the Herbal Recipe Benja Amarit against Highly Invasive Cholangiocarcinoma by Inducing Apoptosis both In Vitro and In Vivo.

Thailand is the country with highest incidence and prevalence of cholangiocarcinoma (CCA) in the world. Due to the frequently late diagnosis that is associated with this disease, most CCA patients are prescribed chemotherapy as a form of treatment. However, CCA is able to resist the presently available chemotherapy, so to the prognosis of this disease is still very poor. In this study, we investigated the anticancer potential of a Thai herbal recipe, Benja Amarit (BJA) against CCA and the relevant mechanisms of action that are involved. We found that BJA inhibited CCA cell viability in a dose-dependent manner, especially in highly invasive KKU-213 cells. The extract induced mitochondrial- and caspase-dependent apoptosis in CCA cells by regulating the nuclear factor-κB (NF-κB) signaling pathway. BJA also triggered autophagy in CCA cells. Nonetheless, the inhibition of autophagy enhanced BJA-induced CCA cell death via apoptosis. An in vivo xenograft model revealed the growth-inhibiting and death-inducing effects of BJA against CCA by targeting apoptosis. However, general toxicity to blood cells, kidneys and the liver, as well as changes in body weight, did not appear. Our findings suggest that the herbal recipe BJA might be used as a potentially new and effective treatment for cholangiocarcinoma patients.

Extracts of Bridelia ovata and Croton oblongifolius induce apoptosis in human MDA­MB­231 breast cancer cells via oxidative stress and mitochondrial pathways.

Breast cancer is the most common type of cancer and is also the second leading cause of cancer­associated death in women worldwide. Thus, there is an urgent requirement for the development of effective treatments for this disease. Bridelia ovata and Croton oblongifolius are herbs used in Thai traditional medicine that have been used to treat various health problems; B. ovata has traditionally been used as a purgative, an antipyretic, a leukorrhea treatment and as a birth control herb. C. oblongifolius has been used to increase breast milk production, for post­partum care (where it is used as a hot bath herb), and as a treatment for flat worms and dysmenorrhea. However, there is little research investigating the anticancer properties of these herbs. The present study aimed to investigate the anticancer properties of crude ethyl acetate extracts of B. ovata (BEA) and C. oblongifolius (CEA) in order to explore their underlying mechanisms in breast cancer cell death. The phytoconstituents of the crude extracts of BEA and CEA were studied using gas chromatography­mass spectrometry (GC­MS). GC­MS analysis showed that the primary compound in BEA is friedelan­3­one, and kaur­16­en­18­oic acid in CEA. Cytotoxicity was investigated using an MTT assay, both BEA and CEA showed greater toxicity against MDA­MB­231 breast cancer cells compared with their effect on MCF10A normal epithelial mammary cells. BEA and CEA exerted various effects, including inducing apoptotic cell death, reducing mitochondrial transmembrane potential, increasing the levels of intracellular ROS, activating caspases, upregulating pro­apoptotic and downregulating anti­apoptotic genes and proteins. BEA and CEA were shown to have anticancer activity against breast cancer cells and induce apoptosis in these cells via a mitochondrial pathway and oxidative stress.

Anticancer effects of a traditional Thai herbal recipe Benja Amarit extracts against human hepatocellular carcinoma and colon cancer cell by targeting apoptosis pathways.

ETHNOPHARMACOLOGICAL RELEVANCE: A preparation of Benja Amarit (BJA) has been effectively used in folk medicine to treat diseases related to the liver and colon and forms of cancer for hundreds of years in Thailand. However, there has not been any research on BJA with regard to its anticancer activity against human hepatocellular carcinoma and colon cancer cells. AIM OF THE STUDY: This study was to obtain the scientific supports for the traditional usage in anticancer potential of BJA extracts on hepatocellular carcinoma and colon cancer. MATERIALS AND METHODS: The cytotoxic effects were determined using MTT assay. Apoptosis was quantitated by annexin V-FITC/PI staining. Caspases activities were measured by using specific substrates and colorimetric analysis. The protein expressions were determined by Western blot analysis. Reactive oxygen species (ROS) generation, mitochondrial transmembrane potential, and calcium ion levels were measured by specific fluorescence probes and flow cytometry. The chick embryo chorioallantoic membrane model has been used to study the in vivo anticancer activity. The phytochemical identification was performed by GC-MS and LC-MS. RESULTS: Notably, 95% (BJA-95) and 50% (BJA-50) ethanolic extract of BJA inhibited hepatocellular carcinoma and colon cancer cell viability in a dose-dependent manner. While, the water extract of BJA (BJA-W) was not found to be toxic to both kinds of cancer cell lines. BJA extract induced both the extrinsic and intrinsic or mitochondria-mediated apoptosis pathways. Moreover, BJA-95 caused ROS generation and endoplasmic reticulum stress-mediated apoptosis. The extract exhibited the growth inhibitory effects on cancer cells in vivo. Phytochemical analysis revealed that the major active compounds were piperine, xanthotoxol and dihydrogambogic acid. CONCLUSION: This study is the first to demonstrate anticancer efficiency of BJA extracts on human cancer cells. We consider BJA extract to be a potentially alternative cancer treatment and to be a promising candidate in the future development of antitumor agents.

Effect of combined extracts of aged garlic, ginger, and chili peppers on cognitive performance and brain antioxidant markers in Aß-induced rats.

A combination of aged garlic, ginger, and chili peppers extracts (AGC) was studied by high-performance liquid chromatography, 2,2-diphenyl-1-picrylhydrazyl, and ferric-reducing antioxidant assays, and oxidative stress markers were analyzed in Aß1-42-induced rats. The AGC was orally administered to Wistar rats at doses of 125, 250, and 500 mg/kg body weight (AGC125, AGC250, AGC500, respectively) for 64 days. At day 56, Aß1-42 was injected via both sides of the lateral ventricles. The effects of the AGC on spatial and recognition memory were examined using a Morris water maze and novel object recognition tasks. Rats induced with Aß1-42 exhibited obvious cognitive deficits, as demonstrated by their increased escape latency time (ET) and decreased retention time (RT) and percentage of discriminative index (DI). When compared with the control group, all AGC-treated rats showed significantly shorter ETs and higher DIs during the 5-min delay testing phase. Rats treated with AGC250 also had significantly longer RTs. Administration of Aß1-42 significantly increased malondialdehyde (MDA) levels and decreased superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) levels in the rat brain homogenate. Pretreatment with the AGC caused significant increases in SOD, GPx, and CAT activities, as well as a significant decrease in MDA in the rat brain homogenates after Aß-induced neurotoxicity. Our results suggested that an AGC may ameliorate cognitive dysfunction in Aß-treated rats due to its role in the upregulation of SOD, GPx, and CAT.

Benefits of aged garlic extract on Alzheimer's disease: Possible mechanisms of action.

Alzheimer's disease (AD) is the most common form of dementia and has become a growing health concern in aging societies. ß-amyloid (Aß) formation in vulnerable brain regions, such as the hippocampus and cerebral cortex is a major neuropathological feature of the disease. Currently, there is no specific drug available for the treatment of AD. However, due to its high antioxidant activity, aged garlic extract (AGE) has been widely used to prevent chronic diseases, such as cancer and cardiovascular disease. A number of studies on the benefits of AGE against cognitive and memory deficits have also been published. This review aimed to summarize the information related to the effects of AGE on learning memory in order to obtain a better understanding of its mechanisms of action. This review also presents an overview of the pathogenesis of AD, and summarizes the main ingredients and neuroprotective effects of AGE against cognitive and learning memory deficits. The mechanisms of action of AGE are also discussed.

Potential of Thai Herbal Extracts on Lung Cancer Treatment by Inducing Apoptosis and Synergizing Chemotherapy.

The incidence of lung cancer has increased while the mortality rate has continued to remain high. Effective treatment of this disease is the key to survival. Therefore, this study is a necessity in continuing research into new effective treatments. In this study we determined the effects of three different Thai herbs on lung cancer. Bridelia ovata, Croton oblongifolius, and Erythrophleum succirubrum were extracted by ethyl acetate and 50% ethanol. The cytotoxicity was tested with A549 lung cancer cell line. We found four effective extracts that exhibited toxic effects on A549 cells. These extracts included ethyl acetate extracts of B. ovata (BEA), C. oblongifolius (CEA), and E. succirubrum (EEA), and an ethanolic extract of E. succirubrum (EE). Moreover, these effective extracts were tested in combination with chemotherapeutic drugs. An effective synergism of these treatments was found specifically through a combination of BEA with methotrexate, EE with methotrexate, and EE with etoposide. Apoptotic cell death was induced in A549 cells by these effective extracts via the mitochondria-mediated pathway. Additionally, we established primary lung cancer and normal epithelial cells from lung tissue of lung cancer patients. The cytotoxicity results showed that EE had significant potential to be used for lung cancer treatment. In conclusion, the four effective extracts possessed anticancer effects on lung cancer. The most effective extract was found to be E. succirubrum (EE).

Kaempferia parviflora Nanosuspension Formulation for Scalability and Improvement of Dissolution Profiles and Intestinal Absorption.

Kaempferia parviflora (KP) is an herbal medicine for enhancement of physical fitness and male sexual function improvement with low oral absorption of the main active compounds, methoxyflavones. The purpose of this study is to optimize the preparation of nanosuspensions of KP extract for enhancing intestinal absorption using antisolvent precipitation technique which is an accessible nanomanufacturing methodology in the small industrial factory. Nanosuspensions were prepared using various types and concentrations of stabilizers. Then, the dry powder of KP nanosuspension was produced by spray drying. Its dissolution rate was determined using USP dissolution apparatus II. The rat everted intestinal sac was tested to confirm the improvement of intestinal absorption of KP nanosuspension. The result showed that 3% sodium lauryl sulfate (SLS) was the optimal condition for covering the nano-size of KP nanosuspension. KP nanosuspensions had particle sizes ranging from 100 to 300 nm with narrow size distribution (PDI < 0.60) and zeta potential at - 58 to - 70 mV. These characteristics were stable at 4°C and 25°C/60%RH for 1-month storage. Its methoxyflavones content also unchanged at 4°C and 25°C/60%RH for 1-month storage. KP nanosuspension released > 80% of the methoxyflavones within 30 min both in 0.1 N HCl and 0.01 M phosphate buffer solution (pH 6.8). Moreover, the developed nanosuspension dramatically improved the rat intestinal absorption about 10-fold. Therefore, the KP nanosuspension was successfully prepared. It has relatively high stability, fast dissolution rate, and high intestinal absorption.

In vivo effect of Kaempferia parviflora extract on pharmacokinetics of acetaminophen.

Kaempferia parviflora is widely used as a food supplement and a herbal medicine for vitalization. Previous study has shown that K. parviflora had CYP2E1 inducer activity. It is likely to affect the metabolism of CYP2E1 substrates such as acetaminophen which is a common household pain relief medicine. This study investigated the possible pharmacokinetic interaction between K. parviflora and acetaminophen in rats. Acetaminophen (100 mg/kg, p.o) was administered to rats for nine consecutive days. On days 4-9, K. parviflora extract (250 mg/kg, p.o) was given to the acetaminophen-treated rats. After co-administration with K. parviflora, the concentrations of acetaminophen during day 5-8 markedly decreased compared with acetaminophen-only group. At day 9, the pharmacokinetic parameters of acetaminophen in the presence of K. parviflora extract also decreased, including area under the concentration-time curve (from 1.68 ± 0.16 to 0.34 ± 0.04 mg.min/mL), the maximum concentration (from 19.10 ± 1.90 to 4.48 ± 0.56 µg/mL), and half-life (from 21.29 ± 1.36 to 10.81 ± 1.24 min). In addition, clearance and the elimination rate constant of acetaminophen were significantly increased (from 0.003 ± 0.000 to 0.006 ± 0.001 L/min and 0.03 ± 0.00 to 0.07 ± 0.01 min-1, respectively) in the presence of K. parviflora extract. These findings provide the data for in vivo herb-drug interaction between K. parviflora extract and acetaminophen. Therefore, the concomitant use of K. parviflora as a food supplement and acetaminophen should occasion therapeutic and safety concerns.

Glucose Tolerance Test and Pharmacokinetic Study of Kaempferia parviflora Extract in Healthy Subjects.

Kaempferia parviflora Wall. ex Baker (KP), Krachaidam in Thai or Thai ginseng, is a herbal medicine that has many potential pharmacological effects. The effect of KP extract on blood glucose level in rodent was reported. This study focused on the oral glucose tolerance test and pharmacokinetic study in healthy volunteers administered with KP extract (90 and 180 mg/day, placebo). The oral glucose tolerance tests were performed at baselines and 28-days of administration. The pharmacokinetics were determined after a single dose administration of the tested products using 3,5,7,3',4'-pentamethoxyflavone (PMF) and 5,7,4'-trimethoxylflavone (TMF) as markers. The results showed that glucose metabolism via oral glucose tolerance test was not affected by KP extract. Blood glucose levels of volunteers at 120 min after glucose loading were able to be returned to initial levels in placebo, KP 90 mg/day, and KP 180 mg/day groups both at baseline and 28-days of administration. The results of the pharmacokinetic study revealed that only TMF and PMF, but not 5,7-dimethoxyflavone (DMF) levels could be detected in human blood. The given doses of KP extract at 90 and 180 mg/day showed a linear dose-relationship of blood PMF concentration whereas blood TMF was detected only at high given dose (180 mg/day). The half-lives of PMF and TMF were 2-3 h. The maximum concentration (Cmax), area under the curve of blood concentration and time (AUC), and time to maximum concentration (Tmax) values of PMF and TMF estimated for the 180 mg/day dose were 71.2 ± 11.3, 63.0 ± 18.0 ng/mL; 291.9 ± 48.2, 412.2 ± 203.7 ng∙h/mL; and 4.02 ± 0.37, 6.03 ± 0.96 h, respectively. PMF was quickly eliminated with higher Ke and Cl than TMF at the dose of 180 mg/day of KP extract. In conclusion, the results demonstrated that KP extract had no effect on the glucose tolerance test. In addition, this is the first demonstration of the pharmacokinetic parameters of methoxyflavones of KP extract in healthy volunteers. The data suggest the safety of the KP extract and will be of benefit for further clinical trials using KP extract as food and sport supplements as well as a drug in health product development.

Preparation of Curcuma comosa tablets using liquisolid techniques: In vitro and in vivo evaluation.

Curcuma comosa (C. comosa) is a Thai medicinal herb that provides numerous pharmacologic activities due to its estrogen-like action. This study aimed to investigate the use of liquisolid technique to prepare tablets containing oleoresin-like crude extract of C. comosa, and to improve the dissolution profiles of its major compounds, diarylheptanoids (DAs). Free flowing powders of C. comosa extract were obtained by adsorption onto solid carriers, microcrystalline cellulose, with colloidal silica as coating material. FTIR results ruled out possible interactions between C. comosa extract and excipients. The results indicated that all of liquisolid tablets met the USP requirements. The release of DAs were significantly increased through liquisolid formulations, compared to crude extract. By decreasing the ratio of carrier to coating from 20 to 10, an improvement in dissolution rate was observed. Addition of additives - namely polymer (polyvinyl pyrrolidone) and/or nonvolatile liquid (propylene glycol) affected tablet properties which involved longer disintegration time and lower DA dissolution. Optimized C. comosa liquisolid formulation was prepared in a carrier to coating ratio of 10 without additives. Stability studies showed that physical properties of liquisolid tablet were not affected by aging, but percent remaining and dissolution profiles of DAs were influenced by storage temperature. In vivo pharmacokinetic behavior of the optimized C. comosa liquisolid tablets was investigated following a single oral administration to rabbits. The results proved that the method used for preparation of liquisolid led to C. comosa tablets with low variation in content uniformity and tablet properties, as well as enhanced dissolution behavior.

Development and evaluation of topical films containing phytoestrogenic diaryheptanoids from Curcuma comosa extract.

RATIONAL: Phytoestrogens have been found to delay signs of skin aging in post-menopausal women, in a way similar to the effects of estrogens. Diarylheptanoids from a rhizome of traditional Thai herb named Curcuma comosa is considered to be a novel class of phytoestrogens. OBJECTIVES: The aims of this study were to prepare effective topical films using mixed types and vary ratios of hydrophobic (Eudragit RL, Eudragit RS, and Eudragit NE) and hydrophilic polymer (hydroxylpropyl methycellulose, HPMC) with Transcutol as a permeation enhancer for delivery of diarylheptanoids to improve signs of skin aging in post-menopausal women. MATERIAL AND METHODS: Topical films were characterized for their physical and mechanical properties. In vitro release, skin permeation and accumulation were evaluated using Franz diffusion cell and the concentrations of diarylheptanoids were determined using high-performance liquid chromatography. RESULTS: The combined formulations between HPMC and Eudragit NE showed the satisfactory physical and mechanical properties, and also provided the highest amount of drug released compared to Eudragit RL and Eudragit RS. When the proportion of HPMC amount in the polymer matrix increased, the cumulative drug release also increased (HPMC: Eudragit NE 6:4 > 5:5 > 4:6). Moreover, they provided a high accumulation of diarylheptanoids within skin when using transcutol as a permeation enhancer. CONCLUSION: The obtained data provided the skin permeation and accumulation behavior of diarylheptanoids, indicating the feasibility of a skin delivery of the C. comosa extract. The developed films might be topically used as an alternative therapy for protection of skin aging in peri and post-menopausal women.

Asiatic acid protects against cognitive deficits and reductions in cell proliferation and survival in the rat hippocampus caused by 5-fluorouracil chemotherapy.

The chemotherapy drug, 5-fluorouracil (5-FU), has been reported to cause cognitive impairments in cancer patients. The drug also reduces cell proliferation and survival in the brain. Asiatic acid (AA) is a triterpene compound found in Centella asiatica that can protect against reduction of neurogenesis in the hippocampus and memory deficits induced by valproic acid (VPA). In the present study, we investigated the preventive effects of AA on the deficits in spatial working memory and cell proliferation and survival caused by 5-FU chemotherapy in a rat model. Male Sprague Dawley rats received 5-FU (5 i.v. injections, 25 mg/kg) on day 8, 11, 14, 17 and 20 of the study. This was co-administered with AA (30 mg/kg, oral gavage tube) either 20 days before receiving 5-FU (preventive), after receiving 5-FU (recovery), or for the entire period of the experiment (throughout). Spatial working memory was determined using the novel object location (NOL) test and hippocampal cell proliferation and survival of dividing cells were quantified using immunohistochemistry. Rats in the 5-FU alone and recovery groups showed memory deficits in the NOL test and reductions in cell proliferation and cell survival in the subgranular zone (SGZ) of the hippocampal dentate gyrus. Rats in the control, AA alone, and both preventive and throughout co-administration groups, however, did not exhibit these characteristics. The results showed that 5-FU chemotherapy impaired memory and reduced cell proliferation and cell survival in the SGZ of the hippocampal dentate gyrus. However, these impairments in the animals receiving 5-FU chemotherapy were restored to control levels when AA was co-administered before and during 5-FU treatment. These data demonstrate that AA can prevent the spatial working memory and hippocampal neurogenesis impairments caused by 5-FU chemotherapy.

Effects of Aged Garlic Extract on Cholinergic, Glutamatergic and GABAergic Systems with Regard to Cognitive Impairment in Aß-Induced Rats.

Alzheimer's disease (AD) has been linked to the degeneration of central cholinergic and glutamatergic transmission, which correlates with progressive memory loss and the accumulation of amyloid-ß (Aß). It has been claimed that aged garlic extract (AGE) has a beneficial effect in preventing neurodegeneration in AD. Therefore, the objective of this study was to investigate the effects of AGE on Aß-induced cognitive dysfunction with a biochemical basis in the cholinergic, glutamatergic, and GABAergic systems in rats. Adult male Wistar rats were orally administered three doses of AGE (125, 250, and 500 mg/kg) daily for 65 days. At day 56, they were injected with 1 µL of aggregated Aß (1-42) into each lateral ventricle, bilaterally. After six days of Aß injection, the rats' working and reference memory was tested using a radial arm maze. The rats were then euthanized to investigate any changes to the cholinergic neurons, vesicular glutamate transporter 1 and 2 proteins (VGLUT1 and VGLUT2), and glutamate decarboxylase (GAD) in the hippocampus. The results showed that AGE significantly improved the working memory and tended to improve the reference memory in cognitively-impaired rats. In addition, AGE significantly ameliorated the loss of cholinergic neurons and increased the VGLUT1 and GAD levels in the hippocampus of rat brains with Aß-induced toxicity. In contrast, the VGLUT2 protein levels did not change in any of the treated groups. We concluded that AGE was able to attenuate the impairment of working memory via the modification of cholinergic neurons, VGLUT1, and GAD in the hippocampus of Aß-induced rats.

Antioxidant and hypoglycemic effects of Momordica cochinchinensis Spreng: (Gac) aril extract on reproductive damages in streptozotocin (STZ)-induced hyperglycemia mice / Efectos antioxidantes e hipoglucémicos del extracto de aril de Momordica cochinchinensis Spreng: (Gac) sobre los daños reproductivos en ratones con hiperglucemia inducida por streptozotocin (STZ)

The aim of present study was to investigate the effect of Momordica cochinchinensis (Gag) aril (GA) aqueous extract on male reproductive system of streptozotocin (STZ)-induced hyperglycemia (HG) mice. GA were extracted with distilled water (DW) and analyzed for in vitro antioxidant capacities. ICR male mice were divided into 7 groups: 1) control, 2) DW, 3) GA 1000 mg/kg BW, 4) HG, 5) HG + glibenclamide, 6 and 7) HG + GA 500 and 1000 mg/kg BW respectively (7 mice/ group). In HG groups, mice were induced by STZ at single dose (150 mg/kg BW). They were treated for consecutive 35 days. All groups were compared for blood glucose levels, weights and histopathologies of reproductive organs, sperm concentration including testicular tyrosine phosphorylation protein patterns by Immuno-Western blotting. The results showed that GA processed antioxidant activities and could significantly decrease blood glucose levels and increase sperm concentration in HG mice. Moreover, GA could change the density of a testicular 70 kDa protein in HG-GA groups. In conclusion, GA extract could improve hyperglycemia and male reproductive damages in STZ-induced HG mice.

El objetivo de este estudio fue investigar el efecto del extracto acuoso de Momordica cochinchinensis (Gag) aril (GA) en el sistema reproductor masculino de ratones hiperglucémicos inducidos por estreptozotocina (STZ). GA fue extraída con agua destilada (DW) y se analizaron las capacidades antioxidantes in vitro. Ratones ICR machos fueron divididos en 7 grupos: 1) control, 2) DW, 3) GA 1000 mg / kg PC, 4) HG, 5) HG + glibenclamida, 6 y 7) HG + GA 500 y 1000 mg / kg PC, respectivamente (7 ratones / grupo). En los grupos HG, los ratones fueron inducidos con STZ en dosis única (150 mg / kg BW). Fueron tratados durante 35 días consecutivos. En todos los grupos se compararon los niveles de glucosa en sangre, los pesos y las histopatologías de los órganos reproductores, la concentración de espermatozoides, incluídos los patrones testiculares de proteínas tirosina fosforilada por Inmuno-Western blot. Los resultados mostraron que GA procesaba actividades antioxidantes y podían disminuir significativamente los niveles de glucosa en sangre y aumentar la concentración de espermatozoides en ratones HG. Además, GA podría cambiar la densidad de una proteína testicular de 70 kDa en grupos HG-GA. En conclusión, el extracto de GA podría mejorar la hiperglucemia y los daños reproductivos masculinos inducidos por STZ en ratones HG.

Antioxidant activity of ginger extract as a daily supplement in cancer patients receiving adjuvant chemotherapy: a pilot study.

PURPOSE: The aim of this study was to examine the antioxidant activity of ginger extract oral supplement in newly diagnosed cancer patients receiving adjuvant chemotherapy compared to placebo. PATIENTS AND METHODS: Newly diagnosed cancer patients receiving moderate-to-high emetogenic potential adjuvant chemotherapy were randomized to receive either a ginger extract (standardized 6-gingerol 20 mg/day) or a placebo 3 days prior to chemotherapy, which they continued daily. Oxidant/antioxidant parameters, including the activities of superoxide dismutase (SOD) and catalase (CAT) and levels of glutathione peroxidase (GPx), total glutathione (GSH/GSSG), lipid peroxidation products detected as malondialdehyde (MDA) and NO2-/NO3-, were measured at baseline and at days 1, 22, 43 and 64 after undergoing chemotherapy. Two-sided statistical analysis, with P < 0.05, was used to determine statistical significance. RESULTS: A total of 43 patients were included in the study: 19 and 24 patients were randomly assigned to the ginger group and placebo group, respectively. Antioxidant activity parameters, including SOD, CAT, GPx and GSH/GSSG, were significantly increased at day 64 in the ginger group compared to those in the placebo group, while MDA and NO2-/NO3- levels were significantly decreased (P < 0.0001). When compared to the baseline, the activities of SOD and CAT and the levels of GPx and GSH/GSSG were significantly higher on day 64 (P = 0.01), while the blood levels of MDA and NO2-/NO3- were significantly decreased (P < 0.01). CONCLUSION: Daily supplement of ginger extract started 3 days prior to chemotherapy has been shown to significantly elevate antioxidant activity and reduce oxidative marker levels in patients who received moderate-to-high emetogenic potential chemotherapy compared to placebo.