The Potential Role of Circulating MicroRNAs in Male Rat Infertility Treated with Kaempferia parviflora

Background. Therapeutic strategies based on herbal plants and diets containing sufficient amounts of antioxidants and essential vitamins are very important factors in treating reproduction and male infertility worldwide. Thus, the aim of this study was to investigate the potential effects of Kaempferia parviflora (KP) on the role of some microRNAs in treated and nontreated infertile rats. In addition, the correlation of expressed microRNAs with sperm count, sperm motility, and sperm viability was identified. The probable use of these microRNAs as a diagnostic marker for predicting the clinical response of infertility to the treatment with KP was also achieved. Methods. In the present study, the potential effects of Kaempferia parviflora (KP) at different doses (140, 280, and 420 mg/kg) for six weeks on male rats with subinfertility were explored. In addition, the effect of KP on the expression of circulating microRNAs and its correlation with the parameters of sexual infertility was identified by performing both in vitro and in vivo assays. In vitro antioxidant activity, sperm functional analysis, serum testosterone, and expression of circulating microRNAs were conducted using colorimetric, ELISA, and real-time RT-PCR analysis, respectively. Results. Kaempferia parviflora (KP) at nontoxic doses of 140–420 mg/kg/day for six weeks significantly improved serum testosterone and epididymal sperm parameters (sperm count, motility, and sperm viability), increased testicular weight, and provided a reduction in the percentage of abnormal spermatozoon in infertile male rats. The expression of miR-328 and miR-19b significantly decreased, and miR-34 significantly increased in infertile rats treated with KP compared to infertile nontreated rats. After six weeks of KP therapy, the change in the expression levels of miRNAs was correlated positively with higher levels of serum testosterone and the measures of epididymal sperm parameters. The respective area under the receiver operating characteristic curve (AUC-ROC) was applied to predict the potential use of miR-328, miR-19b, and miR-34 in the diagnosis of male infertility in treated and nontreated infertile male rats. The data showed that AUC cutoff values of 0.91 for miR-328, 0.89 for miR-19b, and 0.86 for miR34 were the best estimated values for the clinical diagnosis of male rats with infertility. In rats treated with KP for six weeks, AUC cutoff values of 0.76 for miR-328, 0.79 for miR-19b, and 0.81 for miR-34 were the best cutoff values reported for the clinical response of infertility to KP therapy after six weeks. Conclusions. In this study, the improvement of male infertility might proceed via antioxidant and antiapoptotic pathways, which significantly improve spermatogenesis and aphrodisiac properties of males. In addition, the expression of miRNAs, miR-328, miR-34, and miR-19b, in KP-treated and nontreated infertile rats significantly correlated with increased serum testosterone levels and epididymal sperm parameters as well. MicroRNAs, miR-328, miR-34, and miR-19b, might be related to oxidative and apoptotic pathways that proceeded in spermatogenesis. Thus, the use of miRNAs could have a role as diagnostic, therapeutic, and predictive markers for assessing the clinical response of Kaempferia parviflora treatment for six weeks. This may have potential applications in the therapeutic strategies based on herbal plants for male infertility. However, in subsequent studies, the genetic regulatory mechanisms of the expressed miRNAs should be fully characterized.

Qualitative and Quantitative Study of Phyto-Constituents and Antioxidant Potential of Rhizomes of Kaempferia galanga, Kaempferia parviflora and Kaempferia pulchra

Medicinal plants are the potent source of biologically active compounds and have always been of field of interest for the effective chemotherapeutic agents and offering a broad spectrum of activity with greater emphasis on preventive action. The objective of this study was to screen the phytochemicals, estimate the content of phenolic, flavonoids and alkaloids compounds and determines the antioxidant capacity of the rhizomes of Kaempferia galanga Linn, Kaempferia parviflora and Kaempferia pulchra (K. galanga, K. parviflora and K, pulchra, Zingiberaceae). Qualitative analysis of various phytochemical constituents and quantitative analysis of total phenol, flavonoids and alkaloids were determined by the well-known test protocol available in the literature. The ethanolic extract of rhizomes of K. galanga, K. parviflora and K, pulchra was studied for antioxidant activity on different in vitro models namely 1,1-diphenyl, 2-picryl hydrazyl (DPPH) assay and Nitric oxide (NO) radical scavenging method. Phytochemical analysis of ethanolic extract of all three plants revealed the presence of flavonoids, alkaloids, saponins, phenolics, carbohydrate, and tannin. The total phenolic, flavonoids and alkaloids content of ethanolic extract of K. galanga, K. parviflora and K, pulchra rhizomes were 0.813, 1.146; 1.047, 1.237, 0.285; 0.755, 0.822, 0.975/100mg respectively. All extracts showed dose dependent free radical scavenging property in the tested models, which was comparable to that of ascorbic acid. The present study concluded that the crude extract of K. galanga, K. parviflora and K, pulchra rhizomes is a rich source of secondary phytoconstituents which impart significant antioxidant potential. It is expected that the important phytochemical properties recognized by our study in the indigenous medicinal plants will be very useful in the curing of various diseases when taken along with our food.

Evaluating the Effect of Volten VR4® Kaempferia parviflora Extracts on Blood Glucose Levels in Human Type-2 Diabetes Mellitus and Healthy Individual: A Case-Control Study

Background: Volten VR4® capsules contain extract of Kaempferia parviflora (KP), which has been claimed to reduce blood glucose in patients with diabetes. Objective: To evaluate the efficacy of Volten VR4® on healthy individuals and Type-2 Diabetes Mellitus volunteers. Materials and Methods: The extracts of 400 mg capsules of KP were consumed for five days. The fasting and blood glucose levels of 2-hour postprandial were measured at baseline and day 5. A group of 15 healthy young adults between the age of 20- and 30-years old were the control group of the present study, while 12 volunteers aged 35 to 75 years old diagnosed with Type-2 Diabetes Mellitus constituted the experimental group. Data were validated through the Willcoxon and Friedman test statistics and error distribution. Results: It had been shown that the specificity of KP reduced blood glucose levels and has associated with the flavonoids and polymethoxyflavones components. The results showed that consuming VR4® capsules significantly reduced blood glucose, either at the state of fasting or postprandially in diabetic individual. Conclusion: Volten VR4® Kaempferia parviflora extract is safe to be consumed at 400 mg at one time. The study also has shown that the participants are free from adverse reactions and hypoglycaemia. Keywords: Kaempferia parviflora; Blood glucose; Type-2 Diabetes Mellitus; Flavonoids; Polymethoxyflavones; Hypoglycemia and heavy metal elements

Protective Effect of Polymethoxyflavones Isolated from Kaempferia parviflora against TNF-α-Induced Human Dermal Fibroblast Damage

Similar to other organs, the skin undergoes a natural aging process. Moreover, constant direct exposure to environmental stresses, including ultraviolet irradiation, causes the signs of skin aging to appear rather early. Reactive oxygen species (ROS) and inflammatory responses accelerate skin damage in extrinsic aging. In this study, we aimed to investigate the skin protective effects of polymethoxyflavones found in Kaempferia parviflora against oxidative stress and inflammation-induced damage in human dermal fibroblasts (HDFs) stimulated by tumor necrosis factor-α (TNF-α). The experimental data identified 5,7,4′ trimethoxyflavone (TMF) as the most potent constituent in preventing TNF-α-induced HDF damage among the tested compounds and it was not only effective in inhibiting matrix metalloproteinase-1 (MMP-1) production but also in stimulating collagen, type I, and alpha 1 (COLIA1) expression. TMF suppressed TNF-α-stimulated generation of ROS and pro-inflammatory mediators, such as cyclooxygenase-2 (COX-2), interleukin (IL)-1β, and IL-6 in HDFs. TMF also inhibited the pathways regulating fibroblast damage, including mitogen-activated protein kinase (MAPK), activator protein 1 (AP-1), and nuclear factor-kappa B (NF-κB). In conclusion, TMF may be a potential agent for preventing skin aging and other dermatological disorders associated with oxidative stress and inflammation.

Analysis of Lipophilic Antioxidants in the Leaves of Kaempferia parviflora Wall. Ex Baker Using LC–MRM–MS and GC–FID/MS

Lipophilic metabolites such as carotenoids, fatty acids, vitamin K1, phytosterols, and tocopherols are important antioxidants that are used in the cosmetics, foods, and nutraceutical industries. Recently, there has been a growing demand for the use of byproducts (wastes) as a potential source of industrially important compounds. The leaves of Kaempferia parviflora (black ginger) (KP-BG) are major byproducts of KP-BG cultivation and have been reported to contain several bioactive metabolites; however, the composition of lipophilic metabolites in KP-BG leaves has not been examined. In this study, the lipophilic antioxidant profile was analyzed in the leaves of KP-BG plants grown in vitro and ex vitro. Lipophilic compounds, namely carotenoids (80.40–93.84 µg/g fresh weight (FW)), tocopherols (42.23–46.22 µg/g FW), phytosterols (37.69–44.40 µg/g FW), and vitamin K1 (7.25–7.31 µg/g FW), were quantified using LC–MRM–MS. The fatty acid profile of the KP-BG leaves was identified using GC–FID/MS. The content of individual lipophilic compounds varied among the KP-BG leaves. Ex vitro KP-BG leaves had high levels of lutein (44.38 µg/g FW), α-carotene (14.79 µg/g FW), neoxanthin (12.30 µg/g FW), β-carotene (11.33 µg/g FW), violaxanthin (11.03 µg/g FW), α-tocopherol (39.70 µg/g FW), α-linolenic acid (43.12%), palmitic acid (23.78%), oleic acid (12.28%), palmitoleic acid (3.64%), total carotenoids (93.84 µg/g FW), and tocopherols (46.22 µg/g FW) compared with in vitro KP-BG leaves. These results indicate that ex-vitro-grown KP-BG leaves could be used as a valuable natural source for extracting important lipophilic antioxidants.