"Purplish Blue" or "Greenish Grey"? Indigo Qualities and Extraction Yields from Six Species.

Indigo quality is determined by its indigotin content. Another quality indicator is colour. For an evaluation of species, indigo samples from Indigofera tinctoria, Indigofera suffruticosa, Indigofera arrecta, Persicaria tinctoria, Strobilanthes cusia and Wrightia laevis cultivated in Austria and China were visually classified and analysed spectrophotometrically and using a L*a*b* measuring device. In addition to a standardised hot-extraction method without lime, some samples were extracted simulating traditional methods at ambient temperatures using lime. The highest indigotin contents were achieved with Indigofera arrecta (55%, Austria) and Strobilanthes cusia (56%, China). There were no statistically significant differences between the indigo extraction yields of the species cultivated in Austria, but Indigofera arrecta and Persicaria tinctoria had statistically significantly higher indigotin extraction yields than Indigofera tinctoria and Indigofera suffruticosa. From the species extracted in China, Strobilanthes cusia showed higher values in all parameters than Indigofera tinctoria, Indigofera suffruticosa and Wrightia laevis. Compared with the standardised method, the method simulating local practice yielded more indigo but had a lower indigotin content; the indigotin extraction yields did not differ greatly. L*a*b* values enabled precise estimations of the indigotin content, making it an interesting option for quality control, as inexpensive, easy-to-handle L*a*b* measuring instruments have become available.

Clinical outcomes of patients with remitting ulcerative colitis after discontinuation of indigo naturalis.

Indigo naturalis is an effective treatment for ulcerative colitis. However, long-term use of indigo naturalis causes adverse events, such as pulmonary hypertension. The natural history of patients with ulcerative colitis who discontinued indigo naturalis after induction therapy is unknown. Moreover, the clinical features of patients who relapsed within 52 weeks after the discontinuation of indigo naturalis are unclear. This study aimed to assess the clinical outcomes of patients with ulcerative colitis after discontinuation of indigo naturalis and to identify potential markers responsible for relapse. This single-center retrospective study investigated the follow-up of 72 patients who achieved a clinical response 8 weeks after indigo naturalis treatment. We observed relapse in patients with ulcerative colitis after the discontinuation of indigo naturalis. We analyzed the factors predicting long-term outcomes after discontinuation of indigo naturalis. Relapse was observed in 24%, 57%, and 71% of patients at 8, 26, and 52 weeks, respectively. There were no predictive markers in patients who relapsed within 52 weeks after the discontinuation of indigo naturalis. The ulcerative colitis relapse rate after indigo naturalis discontinuation was high. Follow-up treatment is required after the discontinuation of indigo naturalis in patients with ulcerative colitis.

The double-edged sword effect of indigo naturalis.

Indigo naturalis (IN) is a dried powder derived from plants such as Baphicacanthus cusia (Neeks) Bremek., Polygonum tinctorium Ait. and Isatis indigotica Fork. It has a historical application as a dye in ancient India, Egypt, Africa and China. Over time, it has been introduced to China and Japan for treatment of various ailments including hemoptysis, epistaxis, chest discomfort, and aphtha. Clinical and pre-clinical studies have widely demonstrated its promising effects on autoimmune diseases like psoriasis and Ulcerative colitis (UC). Despite the documented efficacy of IN in UC patients, concerns have been raised on the development of adverse effects with long term consumption, prompting a closer examination of its safety and tolerability in these contexts. This review aims to comprehensively assess the efficacy of IN in both clinical and pre-clinical settings, with a detailed exploration of the mechanisms of action involved. Additionally, it summarizes the observed potential toxicity of IN in animal and human settings was summarized. This review will deepen our understanding on the beneficial and detrimental effects of IN in UC, providing valuable insights for its future application in patients with this condition.

Research on different compound combinations of Realgar-Indigo naturalis formula to reverse acute promyelocytic leukemia arsenic resistance by regulating autophagy through mTOR pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: In China, the Chinese patent drug Realgar-Indigo naturalis Formula (RIF) is utilized for the therapy of acute promyelocytic leukemia (APL). Comprising four traditional Chinese herb-Realgar, Indigo naturalis, Salvia miltiorrhiza, and Pseudostellaria heterophylla-it notably includes tetra-arsenic tetra-sulfide, indirubin, tanshinone IIa, and total saponins of Radix Pseudostellariae as its primary active components. Due to its arsenic content, RIF distinctly contributes to the therapy for APL. However, the challenge of arsenic resistance in APL patients complicates the clinical use of arsenic agents. Interestingly, RIF demonstrates a high remission rate in APL patients, suggesting that its efficacy is not significantly compromised by arsenic resistance. Yet, the current state of research on RIF's ability to reverse arsenic resistance remains unclear. AIM OF THE STUDY: To investigate the mechanism of different combinations of the compound of RIF in reversing arsenic resistance in APL. MATERIALS AND METHODS: The present study utilized the arsenic-resistant HL60-PMLA216V-RARα cell line to investigate the effects of various RIF compounds, namely tetra-arsenic tetra-sulfide (A), indirubin (I), tanshinone IIa (T), and total saponins of Radix Pseudostellariae (S). The assessment of cell viability, observation of cell morphology, and evaluation of cell apoptosis were performed. Furthermore, the mitochondrial membrane potential, changes in the levels of PMLA216V-RARα, apoptosis-related factors, and the PI3K/AKT/mTOR pathway were examined, along with autophagy in all experimental groups. Meanwhile, we observed the changes about autophagy after blocking the PI3K or mTOR pathway. RESULTS: Tanshinone IIa, indirubin and total saponins of Radix Pseudostellariae could enhance the effect of tetra-arsenic tetra-sulfide down-regulating PMLA216V-RARα, and the mechanism was suggested to be related to inhibiting mTOR pathway to activate autophagy. CONCLUSIONS: We illustrated that the synergistic effect of different compound combinations of RIF can regulate autophagy through the mTOR pathway, enhance cell apoptosis, and degrade arsenic-resistant PMLA216V-RARα.

Indigo naturalis (Qing dai) for inflammatory bowel disease: A systematic review and meta-analysis.

BACKGROUND: Indigo naturalis (Qing dai) is a traditional therapy reported to be useful in inflammatory bowel disease (IBD), especially for ulcerative colitis. We performed a systematic review of its efficacy and safety in IBD. METHODS: Electronic databases (Pubmed, Embase, and Scopus) were searched on 4th March 2023 to identify reports about the use of indigo naturalis in IBD. We extracted data with respect to clinical response, remission, endoscopic and histological responses, and adverse events with the use of indigo naturalis in IBD. Pooled clinical response rates and remission rates were calculated. The quality of studies was assessed using Joanna-Briggs tools. RESULTS: Nine studies reporting on 299 patients were included. The pooled clinical response rate was 0.796 (95 %CI, 0.7465-0.8379, I2=0), and the clinical remission rate in ulcerative colitis was 0.668 (0.488- 0.809, I2=85.2 %). The pooled relative risk of clinical response was higher in the indigo naturalis group as compared to placebo in the two randomized trials [3.82 (2.04; 7.14, I2=0)]. Except for one reversible pulmonary arterial hypertension case, most reported adverse effects were mild. The endoscopic and histological responses, when reported, suggested that indigo naturalis is effective for ulcerative colitis. The limitations of the systematic review included a small number of randomized studies, reports only from East Asia and a relatively small number of patients, especially for Crohn's disease. CONCLUSION: Indigo naturalis is effective in the treatment of ulcerative colitis. Future studies should evaluate the comparative efficacy with other drugs.

Liquid Chromatography-Tandem Mass Spectrometry-Based Metabolomics Analysis of Indigo Naturalis Treatment of Ulcerative Colitis in Mice.

Ulcerative colitis (UC), often known as UC, is an inflammatory disease of the intestines that has frequent and long-lasting flare-ups. It is unknown precisely how the traditional Chinese drug Indigo Naturalis (IN) heals inflammatory bowel disease, despite its long-standing use in China and Japan. Finding new metabolite biomarkers linked to UC could improve our understanding of the disease, speed up the diagnostic process, and provide insight into how certain drugs work to treat the condition. Our work is designed to use a metabolomic method to analyze potential alterations in endogenous substances and their impact on metabolic pathways in a mouse model of UC. To determine which biomarkers and metabolisms are more frequently connected with IN's effects on UC, liquid chromatography-tandem mass spectrometry analysis of the serum metabolomics of UC mice and normal mice was performed. The outcomes demonstrated that IN boosted the health of UC mice and reduced the severity of their metabolic dysfunction. In the UC model, it was also found that IN changed the way 17 biomarkers and 3 metabolisms functioned.

High-Quality Indigo Naturalis Obtained with Automatic Foam Separation.

Indigo Naturalis is not only an ancient plant dye but also a famous herbal medicine with antibacterial, anti-inflammatory, and anticancer properties. In traditional processes, thousands of manual stirring separate the high-quality Indigo Naturalis from the crude pulp system. However, this method is time-consuming and labor-intensive, resulting in an unstable quality and low yield, which cannot meet the requirements of modern industrial production. In this study, foam-separation technology was used to increase the industrial applicability of high-quality Indigo Naturalis. The process parameters were optimized based on the content of active ingredients, skin irritation effects, and antioxidative stress activity. The results showed that the optimal process of the foam separation achieved the liquid level difference of 40 cm and the foaming intensity of 0.35 MPa. Compared with the original sample, the indigo and indirubin contents in purified Indigo Naturalis were 1.6 and 3 times higher, the total ash content decreased from 86 to 70%, the pH value decreased from 12.18 to 9.71, and the leachate doubled. Animal experiments suggested the significantly reduced irritation (p < 0.01) and enhanced antioxidative stress activity (p < 0.01) of Indigo Naturalis after foam separation. Therefore, the foam-separation equipment developed in this study enabled the refinement of active ingredients in Indigo Naturalis, which greatly improved the production efficiency and quality.

Dissection of scientific compatibility of Chinese medicinal formula Realgar-Indigo naturalis as an effective treatment for promyelocytic leukemia from the perspective of toxicology.

ETHNOPHARMACOLOGICAL RELEVANCE: Realgar-Indigo naturalis formula (RIF), a first-line drug for the treatment of acute promyelocytic leukemia (APL),is also a TCM formula entirely designed based on TCM theories. There have been studies that explain the scientific connotation of the compatibility of RIF from the perspective of pharmacodynamics. However, as one of the arsenic-containing preparations, the safety of realgar is widely concerned, and there has not been systematic studies to explain the scientific connotation of RIF from the perspective of toxicology. AIM OF THIS STUDY: Dissection of scientific compatibility of Chinese medicinal formula Realgar-Indigo naturalis as an effective treatment for promyelocytic leukemia from the perspective of toxicology. MATERIALS AND METHODS: We used normal mice and an APL model to explore (i) the effects of different components on intestinal permeability, (ii) the changes in intestinal flora, and (iii) toxic effects. At the same time, a bionic extraction method was used to study the effects of different components on the dissolution of soluble arsenic in realgar under the acidic environment in the stomach and the alkaline environment in the intestinal tract. RESULTS: Salvia miltiorrhiza Bunge can repair the intestinal mucosal barrier, maintain the homeostasis of intestinal flora, intervene in the dissolution process of realgar, reverse the increase in intestinal permeability and the disturbance of intestinal flora caused by realgar, and reduce toxicity. CONCLUSION: From the perspective of toxicology, we propose new insights into the definition of the roles of each component in the RIF formula, namely realgar is the monarch, Indigo naturalis is the minister, Salvia miltiorrhiza Bungeis the assistant.

Dramatically Improved Severe Pulmonary Arterial Hypertension Caused by Qing-Dai (Chinese Herbal Drug) for Ulcerative Colitis.

Pulmonary arterial hypertension (PAH) is a rare and fatal disease for which some causative drugs have been developed. Qing-Dai is a Chinese herbal drug that is sometimes used as a specific treatment for ulcerative colitis in Asia, including Japan. Here, we report a case of severe Qing-Dai-induced PAH. A 19-year-old woman who has been taking Qing-Dai for 8 months was admitted for exertional dyspnea. Her mean pulmonary artery pressure dramatically improved from 72 to 18 mmHg with Qing-Dai discontinuation and PAH-specific therapy. After 6 years of onset, she had not relapsed with PAH with PAH-specific therapy.

Stability and safety of transbronchial dye mixture for preoperative localization in a porcine model.

OBJECTIVE: For thoracoscopy, the usefulness of a dye mixture of indigo carmine and Lipiodol for localizing lung lesions has been reported. However, little is known about the stability and safety of this dye mixture injected on the visceral pleura through a bronchoscope. METHODS: Porcine models were divided into three groups according to the detection time of the dye mixture: group A with a detection time of 4 h; group B, 8 h; and group C, 24 h. A dye mixture of indigo carmine and Lipiodol (0.5 mL each) was sprayed onto the visceral pleura both in the ventral and dorsal regions via a spray catheter. RESULTS: Twelve markings were created on the visceral pleura of the porcine lung (six ventral and six dorsal) in the six porcine models. At predetermined detection times, all 12 dye markings (100%) were visible on the visceral pleura. The mean longest diameter of the dye marking in the ventral and dorsal regions was 18.8 mm and 24.3 mm, respectively. In groups B and C, pathological changes in the lymphatic system, such as lymphatic dilatations, were found; minimal changes were found in group B, however, these changes with oval-shaped lymphatic cysts and Lipiodol accumulation, were more evident in group C. CONCLUSIONS: The dye mixture of indigo carmine and Lipiodol had reliable stability and visibility. In terms of safety, it may be necessary to check the dye mixture on the lung surface within 8 h.

The use of medicinal plants for the treatment of psoriasis: A systematic review and meta-analysis.

Background Psoriasis is a chronic inflammatory disease that presents as scaly patches on the skin that affects about 3% of the world's population. Adherence to treatment and discrimination against people are common problems, adversely impacts quality of life. Objectives The aim of this study was to investigate the use of medicinal plants as therapeutic adjuvants in the treatment of plaque psoriasis through a systematic review and meta-analysis. Methods A systematic review and meta-analysis of randomized controlled trials in patients with plaque psoriasis was carried out, comparing the efficacy of herbal treatments alone or in association with other therapies. The search was performed in the databases of The Cochrane Library, Lilacs, Medline via PubMed and Embase, only including studies published from 2016 to 2020.The certainty of evidence was assessed using the Grading of Recommendations, Assessment, Development and Evaluation (GRADE) framework. A systematic review and meta-analysis of randomized controlled trials (RCT) in patients with plaque psoriasis was carried out, comparing the efficacy of herbal treatments alone or in association with other therapies. We comprehensively searched the MEDLINE, Embase, Lilacs and Cochrane Library databases, only including studies published from 2016 to 2020. The certainty of evidence was assessed using the GRADE approach. Results Out of 2,268 articles evaluated, only seven RCT were eligible for final analysis. Five of these studies evidenced low risk of bias and a high level of evidence. Limitations Few RCT of medicinal plants. Conclusion This meta-analysis indicates that medicinal plants may be used as topical or oral products, either alone or combined with other forms of treatment. These products have the potential to greatly improve the quality of life of the patient.

Discovery of New Phenylacetone Monooxygenase Variants for the Development of Substituted Indigoids through Biocatalysis.

Indigoids are natural pigments obtained from plants by ancient cultures. Romans used them mainly as dyes, whereas Asian cultures applied these compounds as treatment agents for several diseases. In the modern era, the chemical industry has made it possible to identify and develop synthetic routes to obtain them from petroleum derivatives. However, these processes require high temperatures and pressures and large amounts of solvents, acids, and alkali agents. Thus, enzyme engineering and the development of bacteria as whole-cell biocatalysts emerges as a promising green alternative to avoid the use of these hazardous materials and consequently prevent toxic waste generation. In this research, we obtained two novel variants of phenylacetone monooxygenase (PAMO) by iterative saturation mutagenesis. Heterologous expression of these two enzymes, called PAMOHPCD and PAMOHPED, in E. coli was serendipitously found to produce indigoids. These interesting results encourage us to characterize the thermal stability and enzyme kinetics of these new variants and to evaluate indigo and indirubin production in a whole-cell system by HPLC. The highest yields were obtained with PAMOHPCD supplemented with L-tryptophan, producing ~3000 mg/L indigo and ~130.0 mg/L indirubin. Additionally, both enzymes could oxidize and produce several indigo derivatives from substituted indoles, with PAMOHPCD being able to produce the well-known Tyrian purple. Our results indicate that the PAMO variants described herein have potential application in the textile, pharmaceutics, and semiconductors industries, prompting the use of environmentally friendly strategies to obtain a diverse variety of indigoids.

Localization Technique Using Mixture of Indigo Carmine and Lipiodol of Pulmonary Nodule via Bronchoscopic Navigation.

Background and Objectives: As the number of minimally invasive surgeries, including video-assisted thoracoscopic surgery, increases, small, deeply located lung nodules are difficult to visualize or palpate; therefore, localization is important. We studied the use of a mixture of indigo-carmine and lipiodol, coupled with a transbronchial approach-to achieve accurate localization and minimize patient discomfort and complications. Materials and Methods: A total of 60 patients were enrolled from May 2019 to April 2022, and surgery was performed after the bronchoscopy procedure. Wedge resection or segmentectomy was performed, depending on the location and size of the lesion. Results: In 58/60 (96.7%) patients, the localization of the nodules was successful after localization, and 2/60 required c-arm assistance. None of the patients complained of discomfort during the procedure; in all cases, margins were found to be free from carcinoma, as determined by the final pathology results. Conclusions: We recommend this localization technique using mixture of indigo carmine and lipiodol, in concert with the transbronchial approach, because the procedure time is short, patient's discomfort is low, and success rate is high.

Psoriasis treatment using Indigo Naturalis: Progress and strategy.

ETHNOPHARMACOLOGICAL RELEVANCE: In recent years, there are increasing that the number of patients with psoriasis day by day, and it has become a common disease endangering public health. However, there is no specific cure for psoriasis or control of recurrence. Therefore, it's necessity to seek alternative and efficient therapy, such as Traditional Chinese Medicine (TCM). As a TCM and effective medicine for the treatment of psoriasis, Indigo Naturalis (Baphicacanthus Cusia (Nees) Bremek.) has the effect of clearing heat, detoxifying blood, eliminating spots, reducing fire and calming panic, and it is used in many classical prescriptions for the treatment of psoriasis. AIM OF REVIEW: To review the latest progress and strategies of Indigo Naturalis in the treatment of psoriasis. This manuscript mainly clarifies the traditional medicinal applications, the mechanism of action and application strategies of Indigo Naturalis, and its preparations in the treatment of psoriasis. MATERIALS AND METHODS: Detailed information on Indigo Naturalis was collected from various online databases (PubMed, GeenMedical, Web of Science, Google Scholar, China National Knowledge Infrastructure Database, and National Intellectual Property Administration). RESULTS: This manuscript reviews a great deal of information about how Indigo Naturalis can treat psoriasis through immune cells, signal pathways and disease-related mediators. The mechanism of cymbididae is expounded from the aspects of regulating keratinocyte proliferation and differentiation, regulating inflammatory infiltration of cellular immune system and improving microvascular dilation and hyperplasia in skin lesions. CONCLUSION: The action mechanisms of Indigo Naturalis on psoriasis reflect the characteristics of multiple components, multiple targets, and multiple pathways of Traditional Chinese medicine. However, some pharmacological and clinical research methods are improper, so that the results are difficult to explain at present. Therefore, further in-depth research is needed to provide knowledge in a wider range of areas to confirm the great therapeutic potential of Indigo Naturalis.

[Optimization of extraction process of Children's Qingfei Zhisou Syrup based on pharmacodynamics].

This study determined the extraction rates of indirubin in Indigo Naturalis by ethanol reflux extraction method and water extraction method. The pharmacodynamic study against cough induced by ammonia water in the mouse model and the cough induced by citric acid in the guinea pig model were performed to optimize the extraction process of the sovereign medicinal Indigo Naturalis and the whole prescription of Children's Qingfei Zhisou Syrup. The extraction rate of indirubin by the ethanol reflux method was 51.89%, and indirubin was not detected in the product of water extraction. Two samples of Children's Qingfei Zhisou Syrup prepared with different methods can prolong the incubation period of cough and suppress the frequency of coughs in pharmacodynamic experiments. In terms of prolonging the incubation period of cough, the two samples prepared with different methods had no significant difference. In terms of reducing the frequency of coughs, the high-dose Five kinds of ethanol extracts such as indigo naturalis and three kinds of water extracts such as gypsum had better effect against the citric acid-induced cough of guinea pigs than other samples(P<0.05). The extraction rate of indirubin in Children's Qingfei Zhisou Syrup sample prepared with ethanol was higher than that with water. The two samples of Children's Qingfei Zhisou Syrup prepared with the two methods showed good antitussive effects. The sample prepared with 5 ingredients(including Indigo Naturalis) extracted with ethanol and 3 ingredients(including Gypsum Fibrosum) extracted with water had better alleviation effect on the citric acid-induced cough of guinea pig than the whole water extract sample. In conclusion, the optimum extraction scheme is ethanol extraction for 5 ingredients including Indigo Naturalis in combination with water extraction for 3 ingredients including Gypsum Fibrosum, and the Children's Qingfei Zhisou Syrup produced in this manner has better antitussive efficacy.

Polygonum tinctorium leaves suppress sodium dextran sulfate-induced colitis through interleukin-10-related pathway.

Indigo naturalis, a herbal medicine purified from indigo-containing plants, such as Strobilanthes cusia, Isatis tinctoria, and Polygonum tinctorium, has been reported to be useful in the treatment of ulcerative colitis by activating the aryl hydrocarbon receptor. However, the aryl hydrocarbon receptor pathway causes crucial side effects, such as pulmonary arterial hypertension. Although P. tinctorium is one of the plant derivatives of indigo naturalis, it is not identical to it. To date, the pure leaves of P. tinctorium have not been reported to ameliorate ulcerative colitis. Therefore, we investigated the effect of pure P. tinctorium leaves, which are consumed in some regions, on experimental colitis induced in mice using sodium dextran sulfate. We found that P. tinctorium leaves ameliorated weight loss (P < 0.01) and pathological inflammatory changes in the colon (P < 0.05), enhanced mRNA expression of interleukin-10 (P < 0.05), and decreased expression of tumor necrosis factor-in colonic tissues (P < 0.05), as determined using quantitative real-time reverse transcription polymerase chain reaction. The intraperitoneal administration of an aryl hydrocarbon receptor antagonist did not antagonize the inhibition of mucosal destruction, whereas an anti-interleukin-10 receptor antibody did. These results suggest that P. tinctorium ameliorate sodium dextran sulfate-induced intestinal inflammation via interleukin-10-related pathway, independent of the aryl hydrocarbon receptor pathway. P. tinctorium leaves have the potential to be a new, safe treatment for ulcerative colitis.

Integration of the metabolome and transcriptome reveals indigo biosynthesis in Phaius flavus flowers under freezing treatment.

Background: Indigo-containing plant tissues change blue after a freezing treatment, which is accompanied by changes in indigo and its related compounds. Phaius flavus is one of the few monocot plants containing indigo. The change to blue after freezing was described to explore the biosynthesis of indigo in P. flavus. Methods: In this study, we surveyed the dynamic change of P. flavus flower metabolomics and transcriptomics. Results: The non-targeted metabolomics and targeted metabolomics results revealed a total of 98 different metabolites, the contents of indole, indican, indigo, and indirubin were significantly different after the change to blue from the freezing treatment. A transcriptome analysis screened ten different genes related to indigo upstream biosynthesis, including three anthranilate synthase genes, two phosphoribosyl-anthranilate isomerase genes, one indole-3-glycerolphosphate synthase gene, five tryptophan synthase genes. In addition, we further candidate 37 cytochrome P450 enzyme genes, one uridine diphosphate glucosyltransferase gene, and 24 ß-D-glucosidase genes were screened that may have participated in the downstream biosynthesis of indigo. This study explained the changes of indigo-related compounds at the metabolic level and gene expression level during the process of P. flavus under freezing and provided new insights for increasing the production of indigo-related compounds in P. flavus. In addition, transcriptome sequencing provides the basis for functional verification of the indigo biosynthesis key genes in P. flavus.

Indigo Pulverata Levis (Chung-Dae, Persicaria tinctoria) Alleviates Atopic Dermatitis-like Inflammatory Responses In Vivo and In Vitro.

Atopic dermatitis (AD) is a chronic inflammatory skin disease associated with a type 2 T helper cell (Th2) immune response. The IndigoPulverata Levis extract (CHD) is used in traditional Southeast Asian medicine; however, its beneficial effects on AD remain uninvestigated. Therefore, we investigated the therapeutic effects of CHD in 2,4-dinitrochlorobenzene (DNCB)-induced BALB/c mice and tumor necrosis factor (TNF)-α- and interferon gamma (IFN)-γ-stimulated HaCaT cells. We evaluated immune cell infiltration, skin thickness, and the serum IgE and TNF-α levels in DNCB-induced AD mice. Moreover, we measured the expression levels of pro-inflammatory cytokines, mitogen-activated protein kinase (MAPK), and the nuclear factor-kappa B (NF-κB) in the mice dorsal skin. We also studied the effect of CHD on the translocation of NF-κB p65 and inflammatory chemokines in HaCaT cells. Our in vivo results revealed that CHD reduced the dermis and epidermis thicknesses and inhibited immune cell infiltration. Furthermore, it suppressed the proinflammatory cytokine expression and MAPK and NF-κB phosphorylations in the skin tissue and decreased serum IgE and TNF-α levels. In vitro results indicated that CHD downregulated inflammatory chemokines and blocked NF-κB p65 translocation. Thus, we deduced that CHD is a potential drug candidate for AD treatment.

Evaluation of the treatment of fever, pain and inflammation with Indigofera suffruticosa Miller Leaves Aqueous Extract.

ETHNOPHARMACOLOGICAL RELEVANCE: Indigofera suffruticosa has reports of its popular use for analgesy in several cultures. Besides that, all parts of this plant are used for some medicinal outcome. The leaves are used in teas, decoctions, juices and included in baths for treating fever and inflammatory processes. AIM OF THE STUDY: To evaluate the anti-inflammatory activity mechanisms of I. suffruticosa leave aqueous extract (IsAE). MATERIALS AND METHODS: Phytochemical screening of IsAE was performed by thin layer chromatography. Total flavonoid content was determined and expressed by milligram of quercetin equivalent per gram of extract (mgEQ/g). 50% of the lethal dose that kills animals (LD50) was determined by acute toxicity in mice. Anti-inflammatory activity was evaluated through carrageenan-induced paw edema, peritonitis, and protein denaturation inhibition. Anti-nociceptive potential was evaluated by acetic acid-induced writhing and formalin tests. Antipyretic activity was assessed by yeast-induced fever. RESULTS: Phytochemical analysis revealed the presence of flavonoids and acid gallic in a quantity of 33.9 mg QE/g. Acute toxicity evaluation resulted in a LD50 of 3807.88 mg/kg. For carrageenan-induced paw edema test, IsAE in both doses (20 and 100 mg/kg) reduced the edema in 83.93%. IsAE reduced nitric oxide (NO) production and leucocytes migration to peritonitis inflammation site and at a concentration of 0.5 mg/mL showed also inhibition of protein denaturation similar to indomethacin in the same concentration. IsAE inhibited in 72.60% the number of contortions in writhing test. In formalin test, IsAE was also efficient, but showed results only in the second phase. In addition, the concentration of 100 mg/kg reduced fever significantly. CONCLUSIONS: IsAE proved to be anti-inflammatory, acting in different parts of the inflammation process, confirming its popular use.