Molecular mechanisms of antidiabetic effect of betulinic acid in lotus rhizome.

OBJECTIVES: To explain the bio-physiological mechanisms of the antidiabetic effect of betulinic acid in Lotus rhizome. BACKGROUND: Even though Sri Lankan native medicine uses Lotus rhizome as a medicinal food for diabetes mellitus, its antidiabetic property has not been scientifically explained yet. It is found to compose several medicinally active components with antidiabetic properties, including Betulinic acid. METHODS: A narrator review was conducted with a literature search in PubMed and Google Scholar databases using the search terms "Nelumbo nucifera rhizome", "Lotus rhizome", "phytochemicals", "antidiabetic effect", "hypoglycaemic effect" "Betulinic acid", and "molecular mechanism". RESULTS: The triterpenoid, Betulinic acid exerts its antidiabetic effect via seven bio-physiological mechanisms including, inhibiting α-glucosidase and α-amylase, upregulating the expression of peroxisome proliferator-activated receptor gamma coactivator-1 α, enhancing AS160 protein phosphorylation, stimulating adenosine monophosphate-activated protein kinase activation, stimulating Glucose transporter type 4 synthesis and translocation, inhibiting Protein Tyrosine Phosphatase 1 B activity preventing dephosphorylation of insulin receptor and stimulating Takeda-G-protein-receptor-5 resulting in an increased release of insulin from insulin-containing granulesCONCLUSION: The available scientific knowledge explains that betulinic acid in Lotus rhizome can improve glucose homeostasis contributing to the antidiabetic effect of this root (Tab. 1, Fig. 6, Ref. 29).

Efficacy and safety of Sri Lankan traditional medicine regimen for knee osteoarthritis: study protocol for an open-label, active comparator, randomized controlled trial.

BACKGROUND: Knee osteoarthritis (KOA) is the most common form of arthritis, causing disability and impaired quality of life especially in the elderly. Sri Lankan traditional medicine (STM) is widely used to treat OA, but no clinical trial evaluated on STM regimens for KOA to discuss their safety and efficacy in the treatment. The aim of this study is to compare the efficacy and safety of STM regimen for KOA in comparison to recommended conventional pain management therapy over a period of 8 weeks on relieving the condition. STUDY DESIGN: This is a clinical trial following a protocol-driven open-labeled randomized controlled study enrolling patients with KOA that will be conducted as a single-center trial in the National Ayurveda Teaching Hospital, Sri Lanka. Rasnadvigunabhagasaya herbal decoction (RDBD) and an herbal pill Yoaraja Guggulu were selected as the rescue medication for treating joint disorders. The two Ayurvedic dosage forms will be tested against the non-steroidal anti-inflammatory drugs tab paracetamol and tab ibuprofen as the rescue medication for their safety and efficacy. As test products for external application, oil with an herbal fomentation-Kumburuetaperumkayam Pottani (KAP)-and paste-Sandivadam Lepaya (SVL)-were selected. External applications will be tested against the diclofenac sodium gel and hot water fomentation. KOA patients will be allocated randomly into two arms, and the medications will be given orally for 60 days and externally for 30 days. The primary endpoint is the change in the score on the WOMAC after 08 weeks. WOMAC and KOOS will be recorded and compared between the two arms prior to visiting 1, at the end of 15 days and end of 30 days, and end of the 45 days and end of the second month and 3 months of follow-up. KOOS and WOMAC subscales, a pain disability index, a visual analog scale for pain and sleep quality, and a quality-of-life index are used as secondary outcome measurements. DISCUSSION: This clinical trial will be able to provide evidence-based scientific data on Sri Lankan traditional medicine regimens in the management of KOA. This trial is expected to develop capacity to scientifically evaluate various STM that are claimed to have efficacy in treatment of various disease conditions. TRIAL REGISTRATION: ISRCTN58050062 .

Acute and Subchronic Oral Safety Profiles of the Sudarshana Suspension.

Sudarshana powder (SP) is an Ayurvedic preparation, which contains 53 herbal ingredients along with 50% of Andrographis paniculata and is clinically used with bees honey. This study was aimed to determine the safety profile of the SP, and its novel preparation Sudarshana suspension (SS) on male Wistar rats and tolerance studies were conducted for healthy adult volunteers. Acute and subacute toxicity studies of the SS and hot water extract of SP were assessed in Wistar rats by observing the general behavior, analyzing biochemical and haematological parameters, and pathological observation. Healthy consented adult volunteers (n = 35) of either sex were selected, and tolerance studies of SS were tested by measuring the biochemical and haematological parameters. There were no significant (p > 0.05) changes observed in the treated animals with SS and hot water extract of SP compared with control in body weights, food intake, and water consumption as well as the biochemical and haematological parameters. Histopathological studies revealed no significant (p > 0.05) changes in the liver, heart, and kidney tissues. The experimental results suggest that novel formulation SS was potentially safe for chronic administration in rats, and no significant differences (p > 0.05) were observed in tested parameters on day 3 and day 8 when compared to the day 0 (baseline) values in healthy volunteers. Healthy volunteers did not report any adverse effects or any other complications during the treatment period and the follow-up period. Therefore, it can be concluded that the novel preparation Sudarshana suspension does not cause any significant toxic effects on the blood parameters in animal and human models.

Efficacy and safety of two Ayurvedic dosage forms for allergic rhinitis: Study protocol for an open-label randomized controlled trial.

BACKGROUND: Allergic rhinitis (AR) is an immune response of the nasal mucosa to airborne allergens and involves nasal congestion, watery nasal discharge, itching of the nose, and sneezing. The symptoms of allergic rhinitis may significantly affect a patient's quality of life and can be associated with conditions such as fatigue, headache, cognitive impairment, and sleep disturbances. Various complementary and alternative medicine treatments have been used for this condition in clinical practice. The Ayurveda system of medicine is the most common complementary medicine system practiced in Sri Lanka. The aim of this study is to examine the efficacy and safety of a decoction used in traditional Ayurveda for allergic rhinitis and its ready- to-use freeze dried formulation in comparison to an antihistamine over a period of 4 weeks on relief of symptoms in allergic rhinitis. STUDY DESIGN: This is a three-arm, open-label, non-inferiority, randomized controlled clinical trial enrolling patients with AR. Tamalakyadi decoction containing 12 ingredients (TMD12), used in traditional Ayurveda and its freeze-dried formulation are the test products. The efficacy and safety of the two Ayurvedic dosage forms will be tested against the antihistamine loratadine. Patients with symptoms of AR will be allocated randomly into the three arms after a 1-week run-in period and the medications will be given orally for 28 days. Total Nasal symptom Score (TNSS) of the patients will be used as the primary efficacy endpoint. TNSS will be recorded and compared between the three arms prior to visit 1, at the end of 28 days, and end of the first and second months of follow-up. Symptom scores of daytime nasal symptoms, night time nasal symptoms, non-nasal symptoms and health-related quality of life questionnaire are used as secondary end points. DISCUSSION: This clinical trial will be able to provide evidence-based scientific data on Ayurvedic dosage form, TMD12, and the freeze-dried formulation in the treatment of allergic rhinitis. This trial is expected to develop capacity to scientifically evaluate various Ayurvedic treatments that are claimed to have efficacy in treatment of various disease conditions. TRIAL REGISTRATION: ISRCTN18149439 (6 May 2019).

Comparative Phytochemical Analysis and Antioxidant Activities of Tamalakyadi Decoction with Its Modified Dosage Forms.

BACKGROUND AND OBJECTIVE: Tamalakyadi decoction (TD) is a classical formulation mentioned in authentic traditional medicine text Sarasankshepaya under nasal diseases and used as a remedy for allergic rhinitis. It consists of 12 plant ingredients. Decoction preparations are widely used in Sri Lankan traditional system and considered effective and safe for treating many disorders. However, decoctions have to be used only in fresh state due to shorter shelf life and loss of stability. This decoction preparation method leads to decreasing the patient compliance and is also time consuming. Hence, the objective of the present study was to convert TD to consumer friendly novel dosage form, namely, freeze dried, spray dried, and traditional ganasara forms. METHODOLOGY: Therefore, we compared the phytochemical constituents and antioxidant activities of TD with its modified dosage forms. The chemical comparison of four dosage forms comprises phytochemical screening, TLC and HPTLC fingerprint profiles and the antioxidant activities by DPPH free radical scavenging activity, Ferric reducing antioxidant power (FRAP), total polyphenol content (TPC), and total flavonoid content (TFC). RESULTS: Phytochemical screening revealed the presence of alkaloids, saponins, tannins, steroids, flavonoids, phenols, and terpenoids in all dosage forms. However, the saponins, alkaloids, flavonoids, terpenoids, and steroids were more prominent in TD and freeze dried preparation than the other two preparations. HPTLC fingerprint pattern of freeze dried dosage was more similar with HPTLC fingerprint pattern of TD in terms of number of peaks and their intensity compared to that of spray dried and ganasara dosage forms. Antioxidant activities such as DPPH, FRAP, TPC, and TFC were higher in decoction and freeze dried preparation than in spray dried and ganasara preparation. CONCLUSION: Freeze dried TD is the most suitable ready to use preparation having similar chemical properties and antioxidant activities to TD.

Evaluation of the In Vitro and In Vivo Antioxidant Potentials of Sudarshana Powder.

Sudarshana powder (SP) is one of the most effective Ayurveda powder preparations for paediatric febrile conditions. The objective of the present study was to evaluate the in vitro and in vivo antioxidant potentials of SP. The in vitro antioxidant effects were evaluated using ABTS radical cation decolourization assay where the TROLOX equivalent antioxidant capacity (TEAC) was determined. The in vivo antioxidant activity of SP was determined in Wistar rats using the Lipid Peroxidation (LPO) assay in serum. The in vitro assay was referred to as the TROLOX equivalent antioxidant capacity (TEAC) assay. For the in vivo assay, animals were dosed for 21 consecutive days and blood was drawn to evaluate the MDA level. The in vitro antioxidant activity of 0.5 µg of SP was equivalent to 14.45 µg of standard TROLOX. The percentage inhibition against the radical formation was 50.93 ± 0.53%. The SP showed a statistically significant (p < 0.01) decrease in the serum level of thiobarbituric acid-reactive substance in the test rats when compared with the control group. These findings suggest that the SP possesses potent antioxidant activity which may be responsible for some of its reported bioactivities.

Inhibitory action on the production of advanced glycation end products (AGEs) and suppression of free radicals in vitro by a Sri Lankan polyherbal formulation Nawarathne Kalka.

BACKGROUND: Advanced glycation end products (AGEs) and free radicals are inflammatory mediators and are implicated in many diseases such as diabetes, cancer, rheumatoid arthritis etc. Multi targeted poly herbal drug systems like Nawarathne Kalka (NK) are able to quench the overall effect of these mediators as they contain good combinations of phytochemicals that have least side effects in contrast to modern medicinal drugs. The objectives of this study were to evaluate phytochemical composition, free radical scavenging activity, cytotoxicity and the inhibitory action on the formation of AGEs by aqueous extract of NK. METHODS: Total phenolic content (TPC) and total flavonoid content (TFC) were determined using Folin ciocalteu method and aluminium chloride assay respectively. Free radical scavenging activity was assessed by DPPH radical scavenging assay (DRSA), phosphomolybdenum reduction antioxidant assay (PRAA) and nitric oxide (NO) scavenging assay. Brine Shrimp Lethality (BSL) bioassay was performed as preliminary screening for cytotoxic activity. Inhibitory action on AGE formation was evaluated using fructose mediated glycation of bovine serum albumin using fluorescence spectroscopic method. RESULTS: The TPC and TFC were 75.1 ± 3.0 mg/g gallic acid equivalents and 68.7 ± 7.8 mg/g epigallocatechin gallate equivalents. The DRSA yielded EC50 of 19.15 ± 2.24 µg mL(-1) for NK. DRSA of NK extract was greater than butylated hydroxy toluene (EC50 = 96.50 ± 4.51 µg mL(-1)) but lesser than L-ascorbic acid (EC50 = 5.60 ± 0.51 µg mL(-1)). The total antioxidant capacity of NK as evidenced by PRAA was 106.4 ± 8.2 mg/g L-ascorbic acid equivalents. NK showed EC50 value of 99.3 ± 8.4 µg mL(-1) in the NO scavenging assay compared to the standard ascorbic acid (EC50 = 7.3 ± 0.3 µg mL(-1)). The extract indicated moderate cytotoxic activity in the BSL bioassay. The extract showed effective inhibitory action on the formation of AGEs with EC50 values of 116 ± 19 µg mL(-1), 125 ± 35 µg mL(-1) and 84 ± 28 µg mL(-1) in data obtained over three consecutive weeks respectively. Comparatively the reference standard, aminoguanidine at a concentration of 500 µg mL(-1) demonstrated 65 % inhibition on the formation of AGE after one week of sample incubation. CONCLUSIONS: The results proved the potential of NK as a free radical scavenger, moderate cytotoxic agent and an inhibitor on the formation of advanced glycation end-products.

Effect of Sri Lankan traditional medicine and Ayurveda on Sandhigata Vata (osteoarthritis of knee joint).

Reported case was a 63-year-old female with end-stage osteoarthritis (OA) (Sandhigata Vata) of the left knee joint accompanied by exostoses. Radiology (X-ray) report confirmed it as a Kellgren-Lawrence grade III or less with exostoses. At the beginning, the Knee Society Rating System scores of pain, movement and stability were poor, and function score was fair. Srilankan traditional and Ayurveda medicine treatment was given in three regimens for 70 days. After 70 days, external treatment of oleation and 2 capsules of Shallaki (Boswellia serrata Triana and Planch) and two tablets of Jeewya (comprised of Emblica officinalis Gaertn., Tinospora cordifolia [Willd.] Millers. and Terminalia chebula Retz.), twice daily were continued over 5 months. Visual analogue scale for pain, knee scores in the Knee Society online rating system and a Ayurveda clinical assessment criteria was used to evaluate the effects of treatments in weekly basis. After treatment for 70 days, the Knee Society Rating System scores of pain, movement and stability were also improved up to good level and function score was improved up to excellent level. During the follow-up period, joint symptoms and signs and the knee scores were unchanged. In conclusion, this OA patient's quality of life was improved by the combined treatment of Sri Lankan traditional medicine and Ayurveda.

Functional herbal food ingredients used in type 2 diabetes mellitus.

From many reports it is clear that diabetes will be one of the major diseases in the coming years. As a result there is a rapidly increasing interest in searching new medicines, or even better searching prophylactic methods. Based on a large number of chemical and pharmacological research work, numerous bioactive compounds have been found in functional herbal food ingredients for diabetes. The present paper reviews functional herbal food ingredients with regards to their anti-diabetic active principles and pharmacological test results, which are commonly used in Asian culinary system and medical system and have demonstrated clinical or/and experimental anti-diabetic effectiveness. Our idea of reviewing this article is to give more attention to these functional food ingredients as targets medicinal foods in order to prevent or slow down the development of type 2 diabetes mellitus.

Anti-inflammatory effects of Clematis chinensis Osbeck extract(AR-6) may be associated with NF-κB, TNF-α, and COX-2 in collagen-induced arthritis in rat.

The root of Clematis chinensis Osbeck has been used widely in rheumatoid arthritis in Chinese traditional medicine, and AR-6 is a triterpene saponin isolated from it. In this present study, we investigated the in vivo effects of oral AR-6 in chronic rat with collagen-induced arthritis (CIA) and possible molecular mechanism. CIA was induced by immunizing 56 female Sprague-Dawley (SD) rats with chicken typeIIcollagen (CII). Following eighteen days, the immunization rats with CIA were treated with AR-6 (32, 16, 8 mg/kg), cyclophosphamide (7 mg/kg), and TGP (Total Glucosides of Paeonia) (180 mg/kg) for 7 days, and rats without CIA were given the same volume of purified water. TNF-α and IL-1ß levels in peripheral blood will be measured by ELISA, and Western blot analysis will be used to detect the expression of NF-κB p65 subunits, TNF-α and COX-2, in synovial membrane. We found that therapeutic treatment with AR-6 markedly improves the paw swelling and histopathological changes. Moreover, the serum levels of pro-inflammatory cytokines TNF-α and IL-1ß were markedly lowered, and the expression of NF-κB p65 subunits, TNF-α and COX-2, in the synovial membrane of CIA rats was significantly inhibited in the AR-6-treated groups. These results enable to prove that AR-6 has a potential anti-inflammatory effect in CIA rats, and its mechanism may relate to the inhibition of the expression of NF-κB p65 subunits, TNF-α and COX-2.

Ex vivo and in vivo effect of Chinese herbal pill Yi Shen Juan Bi (YJB) on experimental arthritis.

AIM OF THIS STUDY: Chinese herbal pill Yi Shen Juan Bi (YJB), which is a compound herbal drug, has traditionally been used as an anti-rheumatic drug in Chinese local clinics. Thus, we investigated the therapeutic effect of the YJB in rheumatoid arthritis (RA), using Freund's complete induced adjuvant arthritis (AA) in rat model. METHODS: For that purpose, macrophage derived cytokine tumor necrosis factor alpha (TNF-α) and interleukin-1 (IL-1) were measured in ex vivo by enzyme linked immunosorbent assay (ELISA). We also assayed the effect of YJB on peritoneal macrophage derived nitric oxide (NO) in ex vivo by Griess reaction. Prostaglandin E (PGE) in metapedes was assayed by ultraviolet spectrophotometer method. Further synovial Bax level was examined by Western blot analysis. RESULTS: YJB significantly decreased the production of peritoneal macrophages derived TNF-α, IL-1 and NO. YJB also significantly decreased prostaglandin E (PGE) and upregulated the Bax expression in AA rat's synovium. CONCLUSION: YJB is a potential anti-rheumatic agent targeting the inflammatory and immunomodulatory response of macrophages while down regulating the PGE and up-regulating the pro-apoptotic Bax expression. Such characteristics of YJB on AA may be advantageous to the treatment of clinical rheumatoid arthritis.

Immunomodulatory activity of a Chinese herbal drug Yi Shen Juan Bi in adjuvant arthritis.

OBJECTIVE: To investigate the immunomodulating mechanisms of a Chinese herbal medicine Yi Shen Juan Bi (YJB) in treatment of adjuvant arthritis (AA) in rats. MATERIALS AND METHODS: Levels of serum tumor necrosis factor alpha (TNF-alpha) and interleukin-1beta (IL-1beta) were measured by the Enzyme-Linked Immunosorbent Assay (ELISA). Expression of TNF-alpha mRNA and IL-1beta mRNA in synovial cells was measured with the semi-quantitative technique of reverse transcription-polymerase chain reaction (RT-PCR), while caspase-3 was examined by western blot analysis. RESULTS: The administration of YJB significantly decreased the production of serum TNF-alpha and IL-1beta. It also decreased significantly the TNF-alpha mRNA, IL-1beta mRNA, and caspase-3 expression in synoviocytes. CONCLUSIONS: YJB produces the immunomodulatory effects by downregulating the over-activated cytokines, while it activates caspase-3, which is the key executioner of apoptosis in the immune system. This may be the one of the underlying mechanisms that explains how YJB treats the rheumatoid arthritis.