In Vitro and In Vivo Anti-Inflammatory Activity of a Combination of Extracts from Cinnamomum zeylanicum, Alpinia galanga, and Withania somnifera used in Unani Medicine.

Objectives: Continuous and excessive secretion of pro-inflammatory and anti-inflammatory chemicals and cytokines may further deteriorate inflammation. Anti-inflammatory drugs play an imperative role in inhibiting the evolution of inflammatory diseases. As per the Unani doctrine, a holistic treatment approach is used to treat illnesses. Therefore, drugs having different actions are used to achieve the synergic effect. Three drugs (Cinnamomum zeylanicum, Alpinia galanga, and Withania somnifera), which are frequently used in Unani medicine for joint disorders were selected to evaluate the anti-inflammatory activity of the extract derived from them. Methods: We used RAW 264.7 macrophage cells to see the expression of inflammatory markers IL-1ß, IL-6, and TNF-α. Cytotoxic activity was assessed with MTT assay and Nitric Oxide (NO) was evaluated using Griess reagent. Further, anti-inflammatory activity was evaluated in Wistar Albino rats using carrageenan-induced paw oedema and immunohistochemistry assays for Cyclooxygenase-2 (COX-2). All the data were analyzed using ANOVA and Dunnett t test for multiple comparisons. Results: This extract did not show any cytotoxic effect and the gene expression was significantly reduced for IL-1ß, IL-6, and TNF-α in a dose-dependent manner. Further, NO production was also significantly reduced in the test groups. Immunohistochemistry revealed that the test groups had less inflammation as compared to the control group. Conclusion: It may be inferred that the ethanolic extract of the three herbs has strong anti-inflammatory activity in the tested inflammatory models and the extract is safe as it did not show any cytotoxic effects in both in vitro and in vivo conditions.

A randomized controlled clinical trial to evaluate safety and efficacy of a Unani formulation in the management of Kalaf (Melasma).

OBJECTIVES: Kalaf (Melasma) is an acquired facial hypermelanism. It has direct impact on patient's quality of life and leads to development of various personality disorders. Lack of effective treatment and recurrences have drawn the attention of researcher to find alternative treatment. This study aimed to evaluate safety and efficacy of a topical Unani formulation in the management of melasma. METHODS: We conducted a prospective randomized controlled clinical study on the participants diagnosed with melasma. The participants (n=72) randomized into test (n=36) and control (n=36) groups. Sixty participants (n=30 in each group) completed the duration of therapy. The participants of the test group were treated with a classical Unani formulation and control group with hydroquinone 4%. The primary end point was change in mean MASI score and secondary end point was improvement in quality of life after eight weeks of treatment. RESULTS: The Unani formulation reduced 40.5% mean MASI score (17.31 ± 9.58 to 10.28 ± 5.92) in comparison to 32% reduction in mean MASI score (20.58 ± 9.49 to 13.92 ± 7.38) in the control group after eight weeks of treatment. When comparing with baseline the difference in MASI score was found statistically significant in both groups (p<0.05). On intergroup comparison, the change in MASI score between both groups was not statistically significant (p>0.05). In addition, MQOL and DQLI also improved significantly in both groups. CONCLUSIONS: This study concluded that the Unani formulation and the control drug were equally effective and safer in the management of melasma.

A non-inferiority randomized controlled clinical trial comparing Unani formulations and PUVAsol in non-segmental vitiligo.

OBJECTIVES: Greco-Arab medicine is an ancient system of medicine with greater treasure on therapeutics of vitiligo. The trial Unani formulations have not been scientifically explored for their safety and efficacy, but have been repeatedly prescribed by the great Unani physicians in the management of Baras (vitiligo). Hence, these interventions were selected for the trial. METHODS: In this randomized, controlled, open-label clinical trial, 82 participants with non-segmental vitiligo aged 18-40 years were block randomized to either receive Unani interventions or control for 16 weeks. Out of 82 participants, 42 were randomized to the Unani group and 40 were randomized to the control group. The primary outcome measure was change in vitiligo area scoring index (VASI), which was assessed on weeks 4, 8, 12 and 16. The secondary outcome measures included the patient's global assessment on VAS and investigator's global assessment based on photographic evaluation at baseline and after the treatment. Safety parameters included hemogram, LFTs, RFTs, CXR, ECG, urine, and stool examinations, which were evaluated at baseline and after the treatment. RESULTS: The per-protocol analysis was done on 30 participants in each group and the response in Unani group was not inferior to those receiving control group. The mean ± SD of vitiligo area scoring index (VASI) decreased from 4.09 ± 2.87 and 5.50 ± 5.73 at baseline to 3.13 ± 2.20 and 4.29 ± 4.95 at the end of the trial in both the Unani and control groups respectively. CONCLUSIONS: The study inferred that both the interventions are equally effective and well-tolerated in patients with non-segmental vitiligo.

The Efficacy and Safety of Herbal Unani Formulations in Chronic Plaque Psoriasis: A Single-arm Clinical Trial.

Background: Despite there being advanced treatment options, psoriasis remains an incurable and recurring disease. Noteworthy scholars of Unani (Greco-Arab) medicine have proposed many drugs and formulations for psoriasis but the scientific evidence on the same is scarce. Hence, trial formulations were selected for the study. Primary Study Objectives: This study was designed to evaluate the efficacy and safety of two herbal Unani formulations, Ma΄jun Mundi and Qairuti Karnab, in the management of chronic plaque psoriasis (CPP). Methods/Design: This open-label, single-arm clinical trial was conducted on 33 participants, of whom 30 completed the 12-week treatment course. Setting: This study was conducted at the Central Research Institute of Unani Medicine (CRIUM), Hyderabad, Telangana, India, from 01 August 2018 to 25 May 2019. Participants: Participants of any gender aged 18 to 65 years with clinically diagnosed CPP and psoriasis area severity index (PASI) ≥ 10% were included in the trial. Interventions: The participants received 5 g of Ma΄jun Mundi (a semisolid preparation) orally, twice daily with water, followed by the topical application of Qairuti Karnab (a homogenous paste) to cover the lesions over 12 weeks. Outcome Measures: The primary outcome measure was the change in PASI determined pre- and post-trial in terms of mean and percentage reduction. Secondary outcome measures were changes in patient global assessment (PGA) on a 100 mm visual analog scale, investigator global assessment (IGA) on a 6-point scale, and subjective parameters including erythema, induration, scaling, and itchiness. Results: The analysis revealed a significant reduction in the PASI score, with 12 subjects (40%) achieving PASI 75 and 3 subjects (10%) achieving PASI 90. Significant improvements were also observed in secondary outcome measures with no adverse events. Conclusion: The findings of the study indicate that the trial formulations exhibit a notable anti-psoriatic effect without any adverse effects. The formulations are worthy of further evaluation as an alternative treatment for CPP.

Phytochemical investigation, antioxidant and anticancer activities of various Unani drugs.

OBJECTIVES: To analyze the phytochemicals, antioxidant, and anticancer activities on MCF-7 human breast cancer cell line using aqueous, hydro-ethanol, and methanol extracts of different Unani drugs, e.g., Halela Siyah, Aftimoon, Bisfayej, Ustukhudoos, and Kutki. METHODS: The qualitative examination (alkaloids, terpenoids, tannins, and saponins), anticancer activity, and an antioxidant assay of the three different extracts were done by MTT assay and DPPH assay, respectively, using different Unani drugs. RESULTS: The qualitative examination confirmed the substantive presence of phytochemical constituents in all the extracts of these drugs. The Methanolic extract of Halela Siyah had the highest DPPH scavenging activity (91%), while Bisfayej had the lowest (58%). Similarly, the hydro-ethanolic extract showed approximately identical activity for Halela Siyah (89%), Aftimoon (88%), Bisfayej (84%), Kutki (82%), and Ustukhudoos (81%). The aqueous extracts of Halela Siyah (88%) had the highest DPPH scavenging activity, whereas Bisfayej (73%) had the lowest. The methanolic extract of Aftimoon demonstrated the greatest anticancer activity (IC50 - 108), while Aftimoon showed the least activity (IC50 - 316). Halela Siyah (IC50 - 175) and Aftimoon (IC50 - 178) showed substantially the same activity in aqueous extracts. Ustukhudoos hydro-ethanol extracts had the highest (IC50 - 130) activity, whereas Aftimoon had the lowest (IC50 - 204). CONCLUSIONS: In conclusion, our findings evaluated the presence of phytochemicals, good antioxidant activity, and anticancer activity in different extracts of drugs used in this study. The study shows these drugs have potential anticancer activity against breast cancer in MCF-7 cell lines.

Ethno-pharmacology of Asaroon (Asarum europaeum L.) with special reference to Unani System of Medicine.

Asaroon is the rhizome of Asarum europaeum L. and is commonly used in Unani medicines for its various pharmacological actions. It is an evergreen plant with glossy foliage. It belongs to the family of Aristolochiaceae and is native to Europe and the United State of America. Some species of Asaroon have been found in the Eastern Himalayan region. Asaroon has actions like Muharrik-i-A'sab (nervine stimulant), Mudirr-i-Bawl (diuretics), Mudirr-i-Hayd (emmenagogue), Musakkin-i-Alam (analgesic), Mufattit-i-sudad (remove obstructions) and Muqawwi-i-Jigar (hepatotonic), etc. It is used in the management of Humma (fevers), Waja 'al-Mafasil (polyarthritis), Sara (epilepsy), Falij (paralysis), Ihtibas al-Tamth (amenorrhea) and Niqris (gout), etc. as per the Unani system of medicine (USM). It is used as a single herb as well as with a combination of other drugs to manage many diseases. The A. europaeum L. contains volatile oils and flavonoids along with other secondary metabolites. In the Indian market, Valeriana wallichii DC has been sold as Tagar but in some cases, it is sold as Asaroon. It is a clear case of adulteration by replacement of costly foreign drugs with a similar-looking indigenous drug. In this manuscript, we have discussed the Ethno-pharmacology of the A. europaeum L. with special reference to USM and basic differences with V. wallichii DC to show that both drugs are different and their actions and uses are also different from each other.

Nephroprotective effect of Apium graveolens L. against Cisplatin-induced nephrotoxicity.

BACKGROUND: Cisplatin is extensively used in treating cancers, and its primary side-effect is nephrotoxicity. It accumulates in proximal convoluted tubules where it promotes cellular damage by oxidative stress, apoptosis, and inflammation, etc. In Unani medicine, Tukhm-e-Karafs(Apium graveolens L.) (TK) is mentioned in the literature to manage various kidney ailments due to its diuretic and deobstruent activities. OBJECTIVE: To investigate the nephroprotective effects of powder of TK in Cisplatin-induced nephrotoxicity in an animal model and to validate the Unani claim of its nephroprotective action. MATERIAL AND METHODS: In curative protocol, cisplatin (5 mg/kg body weight i.p) was administered on day one and powder of TK (500 and 1000 mg/kg p.o.) from the sixth day onwards for ten days. TK (500 and 1000 mg/kg p.o.) was given for ten days and Cisplatin (5 mg/kg body weight i.p) on day 11 in the protective model. At the end of the study, all the animals were sacrificed, and renal biochemical parameters were determined. KIM-1 level was also investigated in the kidney homogenate in conjunction with histopathological inspection of kidney tissues. RESULTS: Significant increase in serum creatinine and BUN, presence of mononuclear cell infiltration, tubular dilation and vacuolation in renal histopathology, and increased KIM-1 level confirmed the nephrotoxicity due to Cisplatin. TK's administration protects the kidney as suggested by the changes in biochemical renal function, decreased level of KIM-1, and improvement in histopathological changes. CONCLUSION: The result advocated that TK prevented renal injury and maintained normal renal function in both models. It may be due to improved clearance of Cisplatin from kidney tubules and reduction in reactive oxygen species (ROS) produced by the inflammatory response.

The anticancer and free radical scavenging potential of the Unani medicine Agaricus albus L.

OBJECTIVES: Traditional Unani medicine plays a major role in maintaining health in developing countries. For ages, Unani physicians have described many Classical Unani Formulations (CUF) for their anti-cancerous action (Dafi'-i-saratan) in the human population. These formulations contain various individual drugs which also have anticancer activity. The present study was designed to screen and compare cytotoxic and antioxidant properties of different extracts of Agaricus albus L. (A. albus), a drug used in the Unani system of medicine on human breast cancer cell lines (MCF-7). METHODS: Cytotoxic effect was assessed using MTT assay kit and free radical scavenging activity was done by using DPPH assay. Aqueous, hydro-ethanol and methanolic extracts were used at different concentrations. RESULTS: All extracts were shown good antioxidant and cytotoxic activity. Among all extracts, maximum cytotoxic effect was observed in methanolic extract and aqueous extract at the concentration of 1,000 µg against MCF-7 cell line in comparison to paclitaxel. The maximum antioxidant activity was observed in the hydroethanolic extract at the concentration of 1,000 µg against the MCF-7 cell line in comparison to ascorbic acid. CONCLUSIONS: The present study advocates, the anticancer and antioxidant property of A. albus and suggests that this drug may be used as a potential drug for cancer treatment after a successful and detailed preclinical study and clinical trial.

Obesity Potentiates the Risk of Drug-Induced Long QT Syndrome - Preliminary Evidence from WNIN/Ob Spontaneously Obese Rat.

Drug-induced long QT syndrome (DI-LQTS) is fatal and known to have a higher incidence in women rather than in men. Multiple risk factors potentiate the incidence of DI-LQTS, but the actual contribution of obesity remains largely unexplored. Correspondingly, the present study is aimed to evaluate the susceptibility of DI-LQTS in WNIN/Ob rat in comparison with its lean counterpart using 3-lead electrocardiography. Four- and eight-month-old female WNIN/Ob and their lean controls were used for the experimentation. Non-invasive blood pressure measurement and total body electric conductivity (TOBEC) analysis were carried out. After the baseline evaluations, animals were anesthetized with Ketamine (50 mg/kg). Haloperidol (12.5 mg/kg single dose) was administered intraperitoneally and ECG was taken at 0, 10, 20, 30, 60 min, and 24 h time points. Myocardial lystes were used to assess the BNP, protein carbonylation, and hydroxyproline content. Adiposity, as assessed by TOBEC, is higher in obese rats with elevated mean arterial blood pressure. Baseline-corrected QT interval (QTc) is significantly higher in the obese rat with a wider QRS complex. The incidence of PVC and VT are more intense in the obese rat. Haloperidol-induced QT prolongation in obese rats was rapidly induced than in lean, which was observed to remain till 24 h in obese groups while normalized in lean controls. Higher levels of BNP, protein carbonylation, hydroxyproline content, and relative heart weights indicated the presence of cardiac hypertrophy. The study provides preliminary evidence that obesity can be a potential risk factor for DI-LQTS with faster onset and longer subsistence.

A traditional poly-herbal formulation improves cognitive function in C57BL/6 mice / Una formulación tradicional de poli-hierbas mejora la función cognitiva en ratones C57BL / 6

INTRODUCTION: Khamira Gawzaban Ambari Jadwar Ood Saleeb Wala (KGAJOS) is a polyherbal compound Unani Pharmacopoeial formulation described in traditional Unani texts as Muqawwi-e-Aza-e-Raeesa (tonic for brain, heart, liver and stomach). KGAJOS is reported to possess anxiolytic and antidepressant activity in mice. Though it is used clinically for various neurological conditions, preclinical efficacy of this formulation in learning and memory enhancement / improvement is not established. METHOD: KGAJOS was evaluated for cognitive function improvement activity using Morris water maze test in C57BL/6 mice. Piracetam was used as positive control for comparison. Anymaze video tracking software was used for tracking the path of mice in pool as per standard protocol. RESULTS: During probe trial in Morris water maze test, a significant increase in time spent in platform quadrant was observed at 1000 and 1500 mg/kg bw of KGAJOS (p < 0.01 and 0.001, respectively) as well as in piracetam group (p < 0.01) compared to vehicle control. Latency to reach the platform quadrant (escape latency) was significantly reduced (p < 0.001) in piracetam and KGAJOS group at 1000 and 1500 mg/kg bw compared to vehicle control. No change in time spent in platform quadrant and escape latency was observed at 500 mg/kg bw of KGAJOS. CONCLUSIONS: Morris water maze experiment conducted in mice revealed improved learning and memory function of KGAJOS at the dose levels of 1000 and 1500 mg/kg bw whereas 500 mg/kg bw was not found to be effective. Observed efficacy of KGAJOS confirmed the traditional claims and usage of this formulation in conditions associated with cognition and memory

INTRODUCCIÓN: Khamira Gawzaban Ambari Jadwar Ood Saleeb Wala (KGAJOS) es una formulación de Unani compuesto de poliherbal descrito como tónico para el cerebro, corazón, hígado y estómago. Este estudio se realizó para evaluar la eficacia preclínica de KGAJOS en el aprendizaje y la memoria. MÉTODO: Se evaluó la actividad de mejora de la función cognitiva de KGAJOS utilizando la prueba de laberinto de agua de Morris en ratones C57BL / 6. Se utilizó piracetam como control positivo. Se utilizó el software de seguimiento de video Anymaze para rastrear la ruta. RESULTADOS: Durante la prueba de la sonda, se observó un aumento significativo en el tiempo empleado en el cuadrante de la plataforma a 1000 y 1500 mg / kg de peso corporal de KGAJOS (p < 0,01 y 0,001, respectivamente) y en el grupo de piracetam (p < 0,01) en comparación con el control. La latencia para alcanzar el cuadrante de la plataforma (latencia de escape) se redujo significativamente (p < 0,001) en el grupo de piracetam y KGAJOS a 1000 y 1500 mg / kg de peso corporal en comparación con el control. CONCLUSIONES: El experimento del laberinto de agua de Morris reveló una mejora en la función de aprendizaje y memoria con 1000 y 1500 mg / kg de peso corporal de KGAJOS, mientras que 500 mg / kg de peso corporal no fue efectivo. La eficacia observada de KGAJOS confirmó las afirmaciones tradicionales y el uso de esta formulación en condiciones asociadas con la cognición y la memoria

Compensation to clinical trial participants in India: A gap analysis.

The recent amendments notified by the Government of India, for conducting clinical trial, is greatly appreciable as promoting safety and well-being of human subjects. These rules clearly state that medical management of injuries in clinical trials is mandatory, and clinical trial-related injury or death needs to be compensated over and above the medical management. These rules need to be reconsidered for simplification and better understanding of issues regarding compensation. There is a need of clarity at some points which should be discussed with all stakeholders for better understanding of current regulations. In our view, attention must also be given to academic investigators, during discussion to promote availability of cost-effective treatment in India.

Comparative toxicity study on classical and modified version of Jawarish Jalinoos (a traditional Unani formulation) in rats.

BACKGROUND: Jawarish Jalinoos (JJ) is a classical semisolid traditional Unani formulation clinically used for the treatment of weakness of vital organs, liver, and stomach. Although JJ has been widely used clinically for several decades, no scientific report is available for its safety. METHODS: JJ and its sugar-free tablet version (SFJJ; formulated to target diabetic population) were assessed for safety in rats. Ninety-day repeated dose oral toxicity study was performed as per the Organisation for Economic Co-operation and Development Guideline 408. JJ was orally administered at the dose of 2000 mg/kg bw/d, whereas SFJJ was orally administered at the doses of 506 mg/kg body weight (bw)/d, 1012 mg/kg bw/d, and 2024 mg/kg bw/d for 90 days. The animals were periodically observed for clinical signs of toxicity, mortality, morbidity, body weight changes, and feed consumption. At the end of the study, hematology, clinical biochemistry, electrolytes, gross pathology, relative organ weight, and histological examination were performed. RESULTS: Treatment with SFJJ and JJ showed no significant differences in body weight gain, feed consumption, hematology, clinical biochemistry, and serum electrolytes. No gross pathological findings and differences in relative organ weights were observed between control and drug treated rats. Histological examination revealed no toxicologically significant abnormalities related with SFJJ or JJ treatment. CONCLUSION: The 90-day repeated dose oral toxicity study demonstrates that the no observed adverse effect level of SFJJ and JJ is greater than 2024 mg/kg bw/d and 2000 mg/kg bw/d (p.o.) in rats, respectively. Both formulations were found to be safe up to the tested dose levels and experimental conditions, and therefore safe for clinical use as specified in the literature.