Integrative network pharmacology, molecular docking, and dynamic simulation analysis of a polyherbal formulation for potential therapeutic impact on prostate cancer.

Background: Prostate cancer (PCa) remains a significant health concern globally, prompting a continual search for novel therapeutic strategies. In this study, we employed a comprehensive approach combining network pharmacology, molecular docking and dynamic simulation to explore the potential impact of a polyherbal formulation on PCa. Methods: Utilizing comprehensive network pharmacology approaches, we elucidated the complex interactions between the bioactive compounds within the polyherbal formulation and key targets associated with PCa progression, highlighting their multitarget mechanisms through integrated protein‒protein interaction and KEGG pathway analyses. Molecular docking simulation studies were performed to predict the binding affinities and modes of interaction between the identified bioactive compounds and their respective protein targets. Results: Complex connections comprising 486 nodes and 845 edges were found by the compound-target network analysis. Significant interactions were observed, and the average node degree was 4.23. KEGG research revealed that PCa and the PI3K-Akt signalling pathway are implicated in modulating prostate cancer. The Quercetin docking investigations revealed that the binding energies for AR and PIK3R1 were -9 and -9.5 kcal/mol, respectively. Based on the results of the MD simulations, it appears that tiny molecules and proteins have formed stable complexes with low fluctuations. Conclusion: In conclusion, this comprehensive method emphasises the value of network pharmacology in conjunction with molecular docking and dynamic simulation in revealing the anti-PCa therapeutic potential of polyherbal formulations, opening up new possibilities for the creation of efficient anti-cancer medicines.

In Silico Approaches to Polyherbal Synergy: Protocol for a Scoping Review.

BACKGROUND: According to the World Health Organization, more than 80% of the world's population relies on traditional medicine. Traditional medicine is typically based on the use of single herbal drugs or polyherbal formulations (PHFs) to manage diseases. However, the probable mode of action of these formulations is not well studied or documented. Over the past few decades, computational methods have been used to study the molecular mechanism of phytochemicals in single herbal drugs. However, the in silico methods applied to study PHFs remain unclear. OBJECTIVE: The aim of this protocol is to develop a search strategy for a scoping review to map the in silico approaches applied in understanding the activity of PHFs used as traditional medicines worldwide. METHODS: The scoping review will be conducted based on the methodology developed by Arksey and O'Malley and the recommendations of the Joanna Briggs Institute (JBI). A set of predetermined keywords will be used to identify the relevant studies from five databases: PubMed, Embase, Science Direct, Web of Science, and Google Scholar. Two independent reviewers will conduct the search to yield a list of relevant studies based on the inclusion and exclusion criteria. Mendeley version 1.19.8 will be used to remove duplicate citations, and title and abstract screening will be performed with Rayyan software. The JBI System for the Unified Management, Assessment, and Review of Information tool will be used for data extraction. The scoping review will be reported based on the PRISMA-ScR (Preferred Reporting Items for Systematic Reviews and Meta-Analyses extension for Scoping Reviews) guidelines. RESULTS: Based on the core areas of the scoping review, a 3-step search strategy was developed. The initial search produced 3865 studies. After applying filters, 875 studies were short-listed for further review. Keywords were further refined to yield more relevant studies on the topic. CONCLUSIONS: The findings are expected to determine the extent of the knowledge gap in the applications of computational methods in PHFs for any traditional medicine across the world. The study can provide answers to open research questions related to the phytochemical identification of PHFs, criteria for target identification, strategies applied for in silico studies, software used, and challenges in adopting in silico methods for understanding the mechanisms of action of PHFs. This study can thus provide a better understanding of the application and types of in silico methods for investigating PHFs. INTERNATIONAL REGISTERED REPORT IDENTIFIER (IRRID): PRR1-10.2196/56646.

Safety and efficacy of a polyherbal formulation from traditional Persian medicine in patients with calcium kidney stones: A randomized, double-blinded clinical trial.

Background: 10%-15% of the world's population suffers from kidney stones. Nearly 50% increase was observed in diagnosing and treating nephrolithiasis in the last decades. Effective medical treatment for the disease is not yet well established. Moreover, there is an increasing global demand to manage diseases using complementary and alternative medicine. This study aimed to formulate and assess the safety and efficacy of a multi-ingredient formulation from traditional Persian medicine (TPM) known as Mofatet powder in patients suffering from calcium kidney stones. Materials and Methods: The aqueous extract of Mofatet powder was prepared, freeze-dried, and formulated as capsules. 26 patients in the drug group and 25 patients in the placebo group used 500 mg capsules of the drug/placebo twice daily for 5 weeks. Ultrasonography/kidney, ureter and bladder imaging, urine analysis, and biochemical parameters were evaluated before and after the intervention. Results: The imaging results showed a 60.73% decrease (P < 0.001) in stone size in the drug group. Moreover, the urinary calcium decreased (P = 0.02) and the urinary magnesium increased (P < 0.001) in the drug group. No remarkable changes were observed in the placebo group in these parameters. No significant effect was observed in aspartate transaminase, alanine transaminase, serum creatinine, and blood urea nitrogen levels in none of the groups. Conclusion: This study suggests that Mofatet powder was effective in reducing calcium kidney stones size with no potential nephro/hepatotoxicity. After confirming these results in larger clinical trials with longer duration, this formulation can be considered a treatment for nephrolithiasis.

Response surface methodology based development of an optimized polyherbal formulation and evaluation of its anti-diabetic and anti-obesity potential in high-fat diet-induced obese mice.

Background and aim: The seeds of Nelumbo nucifera, Chenopodium quinoa and Salvia hispanica are known as super foods due to their various therapeutic properties. The present study aimed to develop an optimized polyherbal formulation from edible seeds aqueous extract and to evaluate its anti-diabetic and lipase inhibitory effect on diet-induced obese diabetic mice. Experimental procedure: Response surface methodology based various formulations were evaluated for their potent anti-diabetic, lipase-inhibitory and antioxidant activities. Acute toxicity of the best optimized formulation was conducted. The mice were fed a high fat diet for 10 weeks resulting in hyperglycemia and obesity. Oral tolerance tests (sucrose, starch and lipid) of the formulation were performed. The mice were supplemented with different doses (125, 250 and 500 mg/kg) of the formulation for 6 weeks. The body weight and blood glucose level were monitored on a weekly basis. Finally, histological alterations and lipid profiles were analysed. Results and conclusion: The formulation containing equal concentration (1.5 mg/ml) of each seed extract showed maximum bioactivities. The formulation was found to be safe during toxicity assay. The tolerance tests supported the anti-diabetic and anti-obesity effect. Higher dose (500 mg/kg) of the formulation significantly (p < 0.01) lowered elevated fasting blood glucose, lipid indices and ameliorated the histological alterations in liver, kidney and pancreas caused by high fat diet. We demonstrated for the first time that the developed aqueous extract optimized formulation possess anti-diabetic and anti-obesity potential and thus could be used as adjuvant therapy for holistic management of type 2 diabetes mellitus.

Safety evaluation of standardized Ayurvedic formulation-Panchvalkala, by subacute toxicity study.

ETHNOPHARMACOLOGICAL RELEVANCE: Panchvalkala is a conventional Ayurvedic medicine used as a douche in gynecological disorders such as leucorrhea, infertility, and endometriosis. Recently, we have reported the anticancer activity of Panchvalkala aqueous extract (PVaq) in cervical cancer cell lines, SiHa (HPV16+), HeLa (HPV18+), and mouse papilloma models. AIM OF THE STUDY: Here, we have evaluated the safety of the aqueous extract of Ayurvedic formulation, Panchvalkala (PVaq), in Swiss albino mice by performing subacute toxicity study. MATERIALS AND METHODS: Male and female Swiss albino mice (n = 5/sex/group) were gavaged orally with different doses of PVaq for 28 consecutive days. The mice were distributed into six groups: I (vehicle control), II (vehicle control reversal), III (PVaq 250 mg/kg), IV (PVaq 500 mg/kg), V (1000 mg/kg) and VI (1000 mg/kg high dose reversal). Animals were observed periodically to record any clinical signs of toxicity or mortality. After completion of treatment and recovery periods, animals were evaluated for the effect of PVaq on urine parameters, followed by hematological and biochemical parameters. Animals were sacrificed on day 29 for gross observation of vital organs and to study their histopathology. Reversal groups were maintained for further 14 days to observe any delayed onset of toxic side effects or reversal of toxicity, followed by sacrificing the mice on day 43. RESULTS: In the subacute toxicity study, PVaq did not show any significant change in food, water consumption, and body weights. There were no significant alterations in hematology, biochemistry, urine parameters, and histopathology of the analyzed tissues (brain, heart, liver, lung, spleen, thymus, kidney, epididymis/ovaries, and testis/uterus). The parameters were comparable to their respective controls in both the female as well as the male mice groups. Upon macroscopic and microscopic observation of vital organs, no abnormality was detected compared to the respective control groups. CONCLUSION: The subacute toxicity study demonstrated that oral administration of PVaq was safe in female and male Swiss albino mice.

Acute and sub-chronic toxicity of Liberin, an anti-diabetic polyherbal formulation in rats.

BACKGROUND: The polyherbal formulation (PHF) liberin, is known to exert anti-hyperglycemic effects in type 2 diabetes mellitus. Hence, it is important to study the safety profile of PHF in the current study through acute and chronic toxicity evaluation. OBJECTIVES: This research aims to assess the acute and sub-chronic toxicity of PHF in rats. MATERIALS AND METHODS: PHF was administered once orally (1000 mg/kg body weight), and the rats (male and female) were monitored for toxicity signs for a 14-day period. For a 28-day chronic toxicity study, rats were daily administered with PHF dose of 500 mg/kg and 1000 mg/kg body weight. Rats were followed up for mortality, weight changes, and other morbidities. Further haematological, biochemical, and histopathological changes were assessed. RESULTS: No death related to treatment or toxicity signs were recorded in the acute single-dose administration group. The results showed that the PHF was tolerated well up to a dose of 1000 mg/kg body weight. Even at the high dose of 1000 mg/kg body weight, sub-chronic tests did not show any significant difference between the dosed and normal groups. No significant changes were seen in the histopathological analysis of the liver, spleen, and kidney as well as haematological and biochemical parameters in acute, sub-chronic and satellite groups following the administration of PHF. CONCLUSION: The results confirmed that there was no adverse effect of this PHF at the maximum dose of 1000 mg/kg body weight in Wistar rats. Further, no adverse delayed effects related to PHF were observed in the satellite group. Therefore, this PHF appears safe for therapeutic purposes in the Ayurvedic medicinal system.

Optimization of total phenolic content, total flavonoid content and anti-gout properties of polyherbal formulation.

OBJECTIVES: An increase in gout prevalence has drawn attention among society and this situation drives the exploration of more favourable treatment using traditional medicinal plants which are rich in phenolic and flavonoid to avoid the side effects of modern medication. However, there are only few studies regarding the optimization of phytochemicals and anti-gout properties of medicinal plants and their combinations. The objectives of this study were to determine the optimal formulation of Strobilanthes crispus, Orthosiphon stamineus Benth and Stevia rebaudiana with maximum total phenolic and flavonoid contents as well as minimum IC50 of in vitro xanthine oxidase inhibitory activity and to examine their correlations among the formulations. METHODS: Plant extracts from hot water infusion were tested for the total phenolic content, total flavonoid content and enzyme inhibition through Folin-ciocalteu assay, aluminium chloride method and xanthine oxidase inhibition assay, respectively. Simplex-centroid mixture design was applied in this study and 13 polyherbal formulations were generated by Design Expert Software. RESULTS: Linear, special cubic and quadratic models were selected to describe the interaction effect between polyherbal formulations and their responses. Low IC50 value (13.90 µg/mL) of xanthine oxidase activity was found in the binary combination of O. stamineus and S. rebaudiana and this probably related to its high phenolic and flavonoid contents as xanthine oxidase inhibition and phytochemicals were correlated. CONCLUSIONS: The suggested optimal formulation was comprised of 44.26 % O. stamineus and 55.74 % S. rebaudiana and it could be developed as an alternative treatment for gout.

A Saffron based Polyherbal Formulation DuK Prevents Hepatocellular Carcinoma in Male Wistar Rats.

BACKGROUND: Duk is a well-established traditional drug which has been used since time immemorial by Indian practitioners to cure various human ailments. OBJECTIVE: The purpose of this study was to explore the anti-cancer activity and the possible mechanism of Duk against diethylnitrosamine (DEN)-initiated hepatocarcinogenesis. METHODS & RESULTS: We administered Duk at 3 doses, viz., 75, 150, and 300 mg/kg/day, 2 weeks before the DEN and continued it for 16 weeks. After 1 week of DEN recovery, 2-aminoacetylflourine (2-AAF) was administered to promote hepatocarcinogenesis. We found that Duk significantly reduced the DEN and 2-AAF induced phenotypical changes in rats and restored the activities of serum markers. Furthermore, Duk counteracted the oxidative stress induced by carcinogens as observed by restoration in the levels of superoxide dismutase (SOD) and catalase (CAT). Duk significantly diminished the levels of malondialdehyde (MDA) in a dose dependent manner and restored the liver microarchitecture as assessed by histopathological studies. The results of immunohistochemical staining showed that Duk inhibited the DEN-induced decrease in the number of cells positive for Bid and Caspase-9. It also reduces the number of cells positive for Cyclin D. CONCLUSION: Duk significantly protects rat liver from hepatocarcinogenesis by regulating oxidative damage and restoring serum markers. The chemopreventive effect of Duk might be through induction of apoptosis.

Traditional complementary and alternative medicine (TCAM) for diabetic foot ulcer management: A systematic review.

Diabetic Foot Ulcers (DFUs) are a devastating micro-vascular complication of diabetes with an increased prevalence and incidence and high rate of morbidity and mortality. Since antibiotics are frequently used to treat DFU, managing the condition has proven to be extremely challenging and may eventually lead to the development of antibiotic resistance. Scientists from around the world are working to develop an alternative solution to the problem of drug resistance by exploring complementary and alternative medicines that may be obtained from natural sources. Hence, the review aims to comprehensively report the information on the natural treatments and therapy used to manage DFU. All of the information described in the current study was gathered from electronic scientific resources, including Google Scholar, PubMed, Scopus, Science Direct, and Springer Link. Findings from the current review revealed the pre-clinical and clinical utility of 18 medicinal plants, 1 isolated compound, 7 polyherbal formulations including herbal creams, a few micronutrients including vitamins and minerals, insect products such as propolis, honey and, Maggot debridement therapy for the treatment and management of DFU. Natural therapies possess better efficacy, low cost, and shorter duration of treatment when compared with the conventional treatments; hence, all information made available about them is crucial to alter the direction of treatment. Furthermore, the data presented in this review are up to date on the potential efficacy of natural complementary medicines for alleviating DFU problems in in vitro and in vivo tests, as well as clinical studies.

Comparison of Effectiveness of Hiora SG Gel With Triamcinolone Acetonide Gel in Recurrent Aphthous Stomatitis.

Background The oral condition known as recurrent aphthous ulceration (RAU) or recurrent aphthous stomatitis (RAS) is very prevalent. Its etiopathogenesis is unknown; hence, symptomatic therapy is all that can be offered if it manifests clinically. Lesion care aims to minimize discomfort and the frequency of relapses by bringing active illness under local control in the affected area. The current treatment options that may have negative side effects include the use of topical and systemic steroids, antibiotics, cauterization, and laser therapy. Objectives and Importance This study aimed to compare the efficiency of HiOra SG gel with triamcinolone acetonide gel in the management of RAS. Materials and Methods Fifty individuals with RAS were recruited for the trial and randomly assigned to either group I (HiOra SG gel) or group II (0.1% triamcinolone acetonide ointment; Oraways). After each meal for a total of 10 days, those with mouth ulcers were instructed to topically administer the drugs. The clinical data were analyzed by comparing the ulcer severity scores from the first, fifth, and 10th days. Results There was a statistically significant (p = 0.001) reduction in reported pain, pain duration, and overall ulcer severity across all groups. After therapy, however, neither the HiOra gel group nor the triamcinolone group showed any discernible improvement over the other. Conclusion The present study's findings corroborate the efficacy of HiOra SG gel in the treatment of RAS when compared to triamcinolone acetonide gel (0.1%). In this trial, no patients had any negative reactions to HiOra SG gel. In the future, further studies are needed with larger samples to prove its benefits.

Exploring antidiabetic potential of a polyherbal formulation Madhurakshak Activ: An in vitro and in silico study.

Madhurakshak Activ (MA), a commercial polyherbal antidiabetic preparation is known to manage diabetes mellitus (DM) by reducing blood glucose levels. However, lacks systematic mechanistic evaluation for their molecular and cellular mode of actions. In the present study, hydro-alcoholic and aqueous extract of MA were evaluated for their effects on glucose adsorption, diffusion, amylolysis kinetics and transport across the yeast cells using in vitro techniques. Bioactive compounds identified from MA by LC-MS/MS were assessed for their binding potential against DPP-IV and PPARγ via an in silico approach. Our results revealed that the adsorption of glucose increased dose dependently (5 mM -100 mM). Both extracts exhibited linear glucose uptake into the yeast cells (5 mM - 25 mM), whereas glucose diffusion was directly proportional to time (30-180 min). Pharmacokinetic analysis revealed drug-like properties and low toxicity levels for all the selected compounds. Among the tested compounds, 6-hydroxyluteolin (-8.9 against DPP-IV and PPARγ) and glycyrrhetaldehyde (DPP-IV -9.7 and PPARγ -8.5) have exhibited higher binding affinity compared to the positive control. Therefore, the above compounds were further considered for molecular dynamics simulation which showed stability of the docked complexes. Hence, studied mode of actions might produce a concerted role of MA in increasing the rate of glucose absorption and uptake followed by the in silico studies which suggest that the compounds identified from MA may inhibit DPP-IV and PPARγ phosphorylation.

Polyherbal formulation conjugated to gold nanoparticles induced ferroptosis in drug-resistant breast cancer stem cells through ferritin degradation.

Aim: Due to their minimal side effects, the anti-cancer properties of the polyherbal formulation are being investigated. However, due to their low absorption potential, the administration of polyherbal formulations is restricted. Loading the polyherbal formulation into gold nanoparticles enhances the bioavailability of the polyherbal formulation (PHF) accompanied by reducing the concentration of doxorubicin (dox). Ferroptosis is one of the novel pathways that specifically target cancer stem cells due to high ferritin levels. Hence, in the present study, we conjugated polyherbal formulation with gold nanoparticles and studied its effect on inducing ferroptosis in drug-resistant breast cancer cell lines. Materials and methods: PHF and dox conjugated to gold nanoparticles were characterized using FTIR, UV-Vis spectrophotometer, DLS, particle size analyzer, and XRD. The drug entrapment and efficiency studies were performed to assess the biodegradable potential of the synthesized gold nanoparticles. Paclitaxel-resistant breast cancer stem cells were generated, and an MTT assay was performed to evaluate the cytotoxicity potential of AuNP-PHF and AuNP-dox. Scratch assay and clonogenic assay were performed to assess the migration and proliferation of the cells after treatment with chosen drug combinations. The ability of PHF and dox conjugated to gold nanoparticles to induce ferritinophagy was evaluated by RT-PCR. Finally, image analysis was performed to check apoptosis and cellular ROS using inverted fluorescent microscope. The ability to induce cell cycle arrest was assessed by cell cycle analysis using flow cytometer. Results and conclusion: PHF and dox conjugated to gold nanoparticles showed high stability and showed to induce ferritin degradation in drug resistant breast cancer stem cells through ferritin degradation. AuNP-PHF in combination with low dose of AuNP-Dox nanoconjugate could be used as an effective cancer therapeutic agent, by targeting the autophagy necroptosis axis.

The standardized Jamu pahitan, an Indonesian antidiabetic formulation, stimulating the glucose uptake and insulin secretion in the in-vitro models.

Jamu pahitan is a polyherbal formulation commonly used for the traditional management of diabetes in Indonesia and is mainly prepared from Andrographis paniculata (Burm.f.) Nees. It is widely varied in herbal composition for every region has their own plant component addition to the formulation. A version of the formulation used in the greater Surakarta area contained five plant constituents. This study evaluated the in-vitro glucose uptake and insulin secretion stimulatory activities of Jamu pahitan to provide scientific evidence on its efficacy and safety of use. The water and ethanol extracts of three Jamu pahitan formulations were prepared. The total phenolic content (TPC) of the extracts was evaluated by the standard Folin-Ciocalteau method. Their effects on the viability of L6 skeletal muscle and RIN-m5F pancreatic cells were evaluated by a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. The glucose utilized by L6 myotubes treated with Jamu pahitan was assessed indirectly by the glucose oxidase method. The insulin secreted by RIN-m5F treated with the formulation extracts was analyzed by the enzyme-linked immunosorbent assay (ELISA). The correlation between TPC and the profile of safety and efficacy of the formulation was statistically evaluated. The water extracts of Jamu pahitan were safe and exerted significant glucose uptake and insulin secretion stimulatory activity in L6 and RIN-m5F, respectively. The ethanol extracts showed more potent effects than their water counterpart, albeit they exerted cytotoxic effects on the cells at the higher tested concentrations. The formulations at lower concentrations stimulated the proliferation of RIN-m5F. In addition, the TPC was strongly correlated with the glucose uptake and insulin secretion stimulatory activities and also the IC50 of the cells in positive manner. The present study supported the use of Jamu pahitan for the traditional management of diabetes in Indonesia by stimulating glucose uptake in the muscle cells and improving insulin secretion in ß-cells.

Vathasura Kudineer, an Andrographis based polyherbal formulation exhibits immunomodulation and inhibits chikungunya virus (CHIKV) under invitro conditions.

ETHNOPHARMACOLOGICAL RELEVANCE: Chikungunya disease (CHIKD) is caused by the alphavirus, chikungunya virus (CHIKV) and is characterized by acute fever and joint inflammation; the inflammation continues even after clearance of the virus from the system, persisting for several months to years. Currently, there are no modern medicines/vaccines available for its treatment and use of over-the-counter anti-inflammatory generic medicines to relieve symptoms is generally practiced. In India, Indian traditional medicines hold a lot of promise to treat this infection and are routinely used during outbreaks. AIM OF THE STUDY: In the present study, we characterized the phytochemical and physicochemical properties of aqueous and ethanol extracts of the Vathasura Kudineer (VSK), a Andrographis based Siddha polyherbal formulation. Additionally, we evaluated its immunomodulatory and antiviral potential using an in vitro system. MATERIALS AND METHODS: Aqueous and ethanolic extracts of VSK were prepared and their physico and phytochemical properties were obtained by biochemical and biophysical assays, HPTLC and FTIR. The aqueous extracts of VSK and several of its ingredients were evaluated for their cytotoxicity in Vero cells and using the maximum non-toxic concentration (MNTC), were processed further for evaluating their ability to inhibit CHIKV infection in Vero cells. We performed the co-treatment assay with ethanol extract of VSK and several of its ingredients to assess the antiviral activity against chikungunya virus on Vero cells and through pre-treatment assay (anti-adhesive effect), co-incubation assay (virucidal effect) and post-treatment assay (post-entry effect) were evaluated. Further, we tested the aqueous extract of VSK along with some of its ingredients for their immunomodulatory properties. We performed antioxidant and anti-inflammatory assays using LPS-simulated RAW 264.7 cells. For antioxidant capacity of extracts, we performed extra-cellular ABTS radical scavenging activity and intra-cellular effects on ROS generation and SOD activity. We assessed the effect on most important inflammatory mediators like Nitric oxide (NO) and Prostaglandin E2 (PGE2) and pro-inflammatory cytokines like interleukin-1 beta (IL-1ß) and tumor necrosis factor alpha (TNFα). RESULTS: We provided the fingerprint of the phytochemicals of both ethanol and aqueous extracts of VSK that can be used for identification. We observed that ethanol extract was able to inhibit CHIKV infection at MNTC with 48 h of treatment on Vero cells. Its ingredient VSKI-As (Anethum sowa) found to be most effective to show virucidal effect while VSKI-Cs (Clerodendrum serratum) and VSKI-Pn (Pipper nigrum) found to be effective in post-entry effect. VSK was able to show ABTS radical scavenging activity, reduce ROS generation, inhibit the inflammatory mediators (NO and PGE2) and pro-inflammatory cytokines (IL-1ß and TNFα) production in LPS-stimulated RAW 264.7 cells. CONCLUSIONS: We provided the evidence that VSK has both immunomodulatory as well as antiviral potential. It shows virucidal as well as post-entry effects on chikungunya virus. VSK can inhibit pro-inflammatory cytokines, IL-1ß and TNFα production by suppressing the inflammatory mediators, NO and PGE2.

Multimode Assessment of Commercial Polyherbal Formulation: an In Vitro and In Silico Approach.

Antidiabetic polyherbal formulations (APH) are used in management of diabetes mellitus (DM). High glucose levels in DM are related to oxidative stress leading to its associated complications. Therefore, assessing antioxidant activity of various APH might unveil an antioxidant-rich formulation for management of DM and its associated complications. Subsequently selecting an antioxidant assessment method is a challenging aspect, considering various in vitro assays working with diverse mechanism of action. Therefore, present study aims to validate the sensitivity/capacity of different antioxidant assay, thereby assessing the antioxidant potential of 9-APH. Obtained results revealed the ABTS·+ values were higher compared to DPPH+ assay. I-9-HAE (DPPH+: IC50 53.31 µg/ml), NK-HAE (ABTS·+: IC50 2.71 µg/ml), and MN-HAE (FRAP and TAC) exhibited highest antioxidant capacity. A significant correlation was obtained between TPC-DPPH+ (r2: 0.8187****). Furthermore, three APH with better antiradical potential was chosen for various in vitro and in silico method, for validating scientific antidiabetic propensities. Among the tested extracts, I-9-HAE (α-amylase inhibition: IC50 831.84 µg/ml) and MN-HAE (α-glucosidase inhibition: IC50 558.64 µg/ml and antiglycation: IC50 883.74 µg/ml) have showed highest antihyperglycemic and antiglycation properties. Finally, the secondary-metabolites of selected APH were screened through literature search, Lipinski rule, ADMET, and ProTox-II. Subsequently, in molecular docking for the selected 9 secondary metabolites, highest binding affinity was observed in apigenin-7-glucuronide for DPPiv (- 9.6), GLP-1 (- 8.8), NADPH (- 8.7), and HSA (- 9.4). Thus, obtained result proposes synergistic interaction with high antioxidant potential of the selected 3-APH and can be considered an alternative for management of DM, where multiple secondary metabolites exert holistic biological effects. Furthermore, our study also provides data on sensitivity/capacity of different in vitro antioxidant assays.

Beneficial effect of polyherbal formulation in letrozole induced Polycystic ovarian syndrome (PCOS).

Aim of the study: PCOS is an endocrine condition that results in enlarged ovaries with tiny cysts on the margins. The present study aims to investigate the beneficial effect of polyherbal formulation in Letrozole induced PCOS in female Albino Wistar rats. Materials and methods: Acute toxicity study of the polyherbal formulation by administering a single dose (2000 mg/kg) was done in female Albino Wistar rats (20 weeks, 250 g) following OECD guideline 423. For PCOS induced study female Albino Wistar rats were divided into six groups (6 animals/group). Group I (control) was given 0.5% carboxymethylcellulose (CMC) suspension daily as vehicle control. Letrozole (1 mg/kg) was orally administered for 21 days in Group II to VI for induction of PCOS. After induction of PCOS, animals were treated with standard drug (Group III- Clomiphene citrate- 1 mg/kg) and polyherbal tablets (Group IV - 500 mg/kg, Group V- 750 mg/kg, and Group VI - 1000 mg/kg) up to 50 days. Vaginal smears were taken daily to check the estrous cycle. Body weight was measured weekly. Blood samples were withdrawn on 0,21 and 50 days for the determination of fasting blood glucose, lipid profile, LH, FSH, and hormonal levels. Results: Administration of letrozole caused the abnormality in serum sex hormone profile, lipid profile, glucose, and the estrous cycle. The treatment with polyherbal formulation significantly decreased (P < 0.0001) the level of testosterone and improved estradiol and progesterone levels (P < 0.0001). There was a decrease in elevated glucose levels from 71.51 ± 0.15 mg/dl in disease induced group to 57.33 ± 1.90 mg/dl in treatment groups. The triglycerides level was normalized to 33.41 ± 1.81 mg/dl and HDL level was increased to 40.63 ± 1.35 mg/dl in treatment groups. The polyherbal formulation by exerting its beneficial effect also caused the disappearance of the cysts in the ovaries. Conclusion: The polyherbal formulation was found to be effective in PCOS. The effect may be attributed to the individual herbs reported having a significant effect on the pathophysiology of letrozole induced PCOS.

Amelioration of hyperglycemia and hyperlipidemia in a high-fat diet-fed mice by supplementation of a developed optimized polyherbal formulation.

This study evaluated in vivo anti-diabetic and anti-obesity activity of a polyherbal formulation's methanolic extract containing an optimized ratio of edible seeds (Salvia hispanica, Chenopodium quinoa, Nelumbo nucifera). Diet-induced obese mice model (C57BL/6) was developed by feeding the mice a high-fat diet for 10 weeks resulting in hyperglycemia and obesity. Different doses (125, 250 and 500 mg/kg of body weight) of formulation were administered orally daily for 6 weeks. Fasting blood glucose and body weight were monitored throughout the study. At the end of the study, serum parameters were analyzed and histological examinations were performed. There was a significant reduction in fasting blood glucose levels and body weight in animal groups receiving polyherbal formulation. Lipid profile was improved as revealed by a reduction in serum triglycerides and total cholesterol. Histological study showed an improvement in liver, kidney and pancreatic sections of treated mice. High-performance thin layer chromatography was performed to identify the phytochemicals responsible for the above-mentioned bioactivities. The results revealed the presence of flavonoid (rutin) in seeds of N.nucifera and in the polyherbal formulation. For the first time, this study demonstrated the anti-diabetic and anti-obesity potential of the optimized formulation. The formulation can be used as a potential therapy for management of diabesity.

Formulation and evaluation of SGLT2 inhibitory effect of a polyherbal mixture inspired from Ayurvedic system of medicine.

Background and aim: The ingredients viz., Artemisia roxburghiana, Cissampelos pareira, Stephania glabra, Drimia indica, Roylea cinerea, Tinospora sinensis and Curcuma longa of the present formulation are used to treat diabetes in the Indian traditional medical system. Adopting the concept of multiple herbal mixtures for better therapeutic effects from the ancient Ayurvedic text Sarangdhar Samhita, the present study aimed to develop a polyherbal formulation (PHF) of seven herbs and to evaluate its sodium-glucose cotransporter protein-2 (SGLT2) inhibitory effect on type 2 diabetic rats. Experimental procedure: Streptozotocin (STZ) (60 mg/kg) and nicotinamide (NAM) (120 mg/kg) were intraperitoneally administered to induce type 2 diabetes in Wistar rats. The animals were divided into 5 groups viz. normal control, diabetic control, positive control (dapagliflozin at 0.1 mg/kg) and two test groups (PHF at 250 and 500 mg/kg). Various parameters including blood glucose, serum glutamic pyruvic transaminase (SGPT), serum glutamic-oxaloacetic transaminase (SGOT), bilirubin, triglycerides and creatinine were measured. Results and conclusion: The treatment with PHF (250 and 500 mg/kg) showed a significant (p < 0.05) decrease in blood glucose levels by 56.37% and 58.17%, respectively. The levels of SGOT, SGPT and bilirubin were significantly reduced in PHF-fed diabetic rats. Histopathological examination revealed no major changes in the treated groups as compared to the normal control. The molecular docking study showed strong binding of ß-sitosterol, insulanoline, warifteine, dehydrocorydalmine, taraxerol acetate, lupeol, corydalmine and luteolin to SGLT2 protein. The present study concludes that PHF has promising antidiabetic activity via inhibiting SGLT2 protein without showing any adverse effects.

Lipomodulatory and anti-oxidative stress effects of a polyherbal formulation based on garlic and avocado seed extracts on high fat high sucrose diet fed rats.

Objective: To determine antioxidant potentials of Allium sativum and Persea americana seeds extracts and three formulation-based extracts in vitro, and to evaluate the effects of the best formulation on oxidative stress and dyslipidemia on rats fed with high fat and high sucrose diet (HFHSD). Methods: Aqueous extracts of Allium sativum, Persia. americana and three formulations were mixed at various portions (A. s/P. a; w/w): F (1:1), F (3: 1), and F(1:3). They were then tested for their antioxidant potentials in vitro using FRAP, DPPH and NO radicals to identify the best formulation. Four hundred (400) mg/kg b.w. of formulation F(1:1) were administered once daily for 21 days to rats previously fed with HFHSD for 8 weeks. Standard diet, vitamin E, and Atorvastatin were used as controls. After 21 days, body weight, blood glucose, lipid markers, activities of transaminases and markers of the antioxidant systems were assessed. Results: The Formulation F(1:1) showed the best in vitro activity with IC50 values of 6.5 and 2.23 mg/mL respectively for FRAP and DPPH- radical scavenging capacity. HFHSD caused a depletion of antioxidants associated with an increase of pro-oxidants and all the lipid markers except HDL-c Treatment with F(1:1) significantly increased TAC, SOD, and catalase activities, while MDA, protein carbonyls, and NO levels decreased (p < 0.05). Formulation F(1:1) decreased triglycerides (119.88 ± 4.25 mg/dL) and LDL-c (3.78 ± 0.66 mg/dL) levels and significantly increased the HDL-c level: (108.07 ± 6.29 mg/mL). Furthermore, Formulation F(1:1) significantly caused weight loss (2.31%), reduced blood glucose levels (27.38%) and ALT activity. Conclusion: The formulation F(1:1) could be a good candidate for the prevention and treatment of oxidative stress, dyslipidemia and features of metabolic syndrome.

Medha Plus - A novel polyherbal formulation ameliorates cognitive behaviors and disease pathology in models of Alzheimer's disease.

Alzheimer's disease (AD) is a multi-faceted neurodegenerative disorder that leads to drastic cognitive impairments culminating in death. Pathologically, it is characterized by amyloid-ß (Aß) plaques, neurofibrillary tangles and neurodegeneration in brain. Complete cure of AD remains elusive to date. Available synthetic drugs only provide symptomatic reliefs targeting single molecule, hence, are unable to address the multi-factorial aspects in AD pathogenesis. It is imperative to develop combinatorial drugs that address the multiple molecular targets in AD. We show a unique polyherbal formulation of Brahmi, Mandukaparni, Shankhpushpi, Yastimadhu, Kokilaksha and Shunthi called 'Medha Plus' (MP), conventionally used for improving memory and reducing anxiety, was able to ameliorate cognitive deficits and associated pathological hallmarks of AD. Viability assays revealed that MP prevented Aß-induced loss of neurites as well as neuronal apoptosis in cellular models. An array of behavioral studies showed that MP was able to recover AD-associated memory deficits in both Aß-injected rats and 5XFAD mice. Immunohistochemical studies further revealed that MP treatment reduced Aß depositshpi and decreased apoptotic cell death in the hippocampus. Enzymatic assays demonstrated anti-oxidative and anti-acetyl cholinesterase properties of MP especially in hippocampus of Aß-injected rats. An underlying improvement in synaptic plasticity was observed with MP treatment in 5XFAD mice along with an increased expression of phospho-Akt at serine 473 indicating a role of PI3K/Akt signaling in correcting these synaptic deficits. Thus, our strong experiment-driven approach shows that MP is an incredible combinatorial drug that targets multiple molecular targets with exemplary neuroprotective properties and is proposed for clinical trial.