Earthworm extract attenuates silica-induced pulmonary fibrosis through Nrf2-dependent mechanisms.

Silicosis is an occupational pulmonary fibrosis caused by inhalation of silica (SiO2) and there are no ideal drugs to treat this disease. Earthworm extract (EE), a natural nutrient, has been reported to have anti-inflammatory, antioxidant, and anti-apoptosis effects. The purpose of the current study was to test the protective effects of EE against SiO2-induced pulmonary fibrosis and to explore the underlying mechanisms using both in vivo and in vitro models. We found that treatment with EE significantly reduced lung inflammation and fibrosis and improved lung structure and function in SiO2-instilled mice. Further mechanistic investigations revealed that EE administration markedly inhibited SiO2-induced oxidative stress, mitochondrial apoptotic pathway, and epithelial-mesenchymal transition in HBE and A549 cells. Furthermore, we demonstrate that Nrf2 activation partly mediates the interventional effects of EE against SiO2-induced pulmonary fibrosis. Our study has identified EE to be a potential anti-oxidative, anti-inflammatory, and anti-fibrotic drug for silicosis.

Standardization of homeopathic mother tincture of Toxicodendron pubescens and correlation of its flavonoid markers with the biological activity.

BACKGROUND: Standardization and quality control of homeopathic drugs is very challenging. As mother tinctures are derived from complex natural resources, there is a need of systematic evaluation of chemical markers which correlate with the proposed biological activities of mother tinctures. METHODS: In present study, High-Performance Thin-Layer Chromatography (HPTLC) standardization method of homeopathic mother tinctures of Toxicodendron pubescens using quercitrin and rutin as chemical markers is validated and correlations of content of these markers with its anti-inflammatory effects are established. For HPTLC analysis, precoated silica gel plates were used as stationary phase. Two flavonoids, namely quercitrin and rutin were used as markers. Separation was achieved using methylene chloride:methanol:water:glacial acetic acid (15:1.5:1:8 v/v/v) as mobile phase. The developed plates were scanned at 365 nm. RESULTS: It was observed that quercitrin (Rf value 0.63) and Rutin (Rf value 0.41) are well resolved. The minimum detectable concentrations for quercitrin and rutin were 5 ng/spot. The linearity range was between 100 and 2000 ng/spot for both the markers. Subsequently, anti-inflammatory activity of these formulations was determined against carrageenan-induced paw edema in rats, pain threshold determined by electronic Von-Frey apparatus and paw withdrawal latency (PWL) on hot-plate. All the tested formulations of Rhus Tox showed anti-inflammatory and analgesic activity against carrageenan induced paw edema in rats. Quantitative correlation between the content of markers and anti-inflammatory activity of mother tinctures was established. RESULTS: Anti-inflammatory effect as well as effect on paw withdrawal and pain threshold, at third hour after carrageenan injection, correlated with quercitrin and rutin content in the respective formulations. CONCLUSIONS: This study validates a quantitative HPTLC method for standardization of homeopathic mother tincture of Rhus Tox and establishes quercitrin and rutin as markers corresponding its biological activity. Contents of quercitrin and rutin in T. pubescens mother tincture correlates with its anti-inflammatory and analgesic actions and the validated HPTLC method can be used in standardization of homeopathic mother tincture of T. pubescens.

Homeopathic Rhus toxicodendron has dual effects on the inflammatory response in the mouse preosteoblastic cell line MC3T3-e1.

BACKGROUND: Homeopathic remedy Rhus toxicodendron (Rhus tox) is used for several symptoms including skin irritations, rheumatic pains, mucous membrane afflictions, and typhoid type fever. Previously, we reported that Rhus tox treatment increased the cyclooxygenase-2 (COX-2) mRNA expression in primary cultured mouse chondrocytes. METHODS: A preosteoblastic mouse cell line, MC3T3-e1, was treated with different homeopathic dilutions of Rhus tox and the COX-2 mRNA and protein expression was examined using reverse transcriptase-polymerase chain reaction (RT-PCR) and immunoblotting. Additionally, nitric oxide (NO) generation was examined in LPS-induced MC3T3-e1 cells using a Griess reaction assay. RESULTS: Stimulation with different concentrations of Rhus tox increased the expression of Cox2 mRNA, with 30X Rhus tox showing the most prominent increase in mRNA expression. In addition, treatment with 30X Rhus tox significantly increased prostaglandin E2 (PGE2) release compared with other homeopathic dilutions. However, the COX-2 protein expression level differed slightly from its mRNA expression, because the 30C Rhus tox treatment increased COX-2 protein to a greater extent compared with other dilutions. NO generation was dramatically decreased in MC3T3-e1 cells after Rhus tox treatment co-stimulated with lipopolysaccharide. CONCLUSION: Homeopathic dilution of Rhus tox has a dual activity that increases COX-2 expression and decreases NO generation, thus modulating inflammation. Further study is needed to examine the cellular signaling mechanisms that are associated with inflammatory regulation by Rhus tox treatment in greater detail.

[Analysis of liposoluble constituents in Holotrichia diomphalia by GC-MS and investigation their anti-inflammatory and analgesic activities].

OBJECTIVE: To analysis liposoluble constituents of Holotrichia diomphalia by GC-MS and measure their anti-inflammatory and analgesic activities. METHODS: The composition of liposoluble constituents were determined by GC-MS. The dimethylbenzene-induced mice inflammatory models were established. The pain models were obtained by hot plate and acetic acid in mice. RESULTS: Twenty-two components were identified from the petroleum ether extract of Holotrichia diomphalia. The major components were oleic acid, palmitic acid and palmitoleic acid. The petroleum ether extract was able to significantly inhibit the mice ear edema induced by dimethyl-benzene. The pain in mice caused by acetic acid and hot plate were evidently suppressed by the petroleum ether extract. CONCLUSION: The petroleum ether extract of Holotrichia diomphalia has obvious anti-inflammation and analgesic effects.

Modulation of arthritis in rats by Toxicodendron pubescens and its homeopathic dilutions.

BACKGROUND: Toxicodendron pubescens P. Mill (Anacardiaceae) known in homeopathy as Rhus toxicodendron (Rhus tox) is used as an anti-inflammatory medicine in homeopathic practice. In this study, Rhus tox in its crude form and homeopathic dilutions (3cH, 6cH, 30cH, 200cH) was evaluated for effects on Complete Freund's Adjuvant (CFA) induced arthritis in rats. METHOD: We assessed the severity of arthritis through observations including inflammatory lesions, body and organ weight and hematological parameters including C-reactive protein (CRP). Blinded radiological analysis of the affected joints and pain intensity determination was also carried out. RESULTS: Rhus tox protected rats from CFA-induced inflammatory lesions, body weight changes and hematological alterations. Rhus tox protected against radiological joint alterations due to arthritis. Arthritic pain scores were also favorably affected by Rhus tox. All the dilutions of Rhus tox including crude form showed anti-arthritic activity. The maximum protective effect was evident in the crude form at 10mg/kg/day, by mouth. CONCLUSION: This study supports claims in the homeopathic literature on the role of Rhus tox and its ultra dilutions in the treatment of arthritis and associated pain. Further study is needed to explain this anti-arthritic effect of Rhus tox.

Application of "spider-web" mode in discovery and identification of Q-markers from Xuefu Zhuyu capsule.

BACKGROUND: The selection of quality control indicators in a complex system is a key scientific issue for the study of Chinese materia medica (CMM), which is directly related to its safety and efficacy. In order to scientifically understand and control the quality of CMM, quality marker (Q-marker) has been recently raised as a new concept, which provided a novel research idea for the quality control and evaluation of CMM. PURPOSE: By a new and integrated "spider-web" mode, Q-markers of Xuefu Zhuyu capsule (XZC) were comprehensively uncovered, conducing to great improvement of quality control of XZC. METHODS: Mainly established by three dimensions derived from six variables including content, stability and activity, "spider-web" mode was constructed to evaluate Q-marker property of candidate compounds by taking regression area of the tested compounds into account. RESULTS: The candidate compounds with larger regression area were preferentially adopted as Q-markers, which should possess the satisfactorily integrated properties of content, stability and activity. Six compounds, naringin, isoliquiritin, paeoniflorin, protocatechuic acid, neohesperidin and ferulic acid, were identified and preferred as Q-markers of XZC. CONCLUSION: Based on "spider-web" mode, Q-markers from Xuefu Zhuyu capsule were successfully screened, which would substantially perform quality control of XZC and prove the feasibility of "spider-web" mode in solving the selection of quality control indicators from compound formulae.

Immunomodulatory activity of Toxicodendron pubescens in experimental models.

BACKGROUND: Toxicodendron pubescens is a botanical name of Rhus toxicodendron (Rhus tox). This plant is widely used in its homeopathically diluted form in the treatment of inflammatory and edematous conditions. In this study, various dilutions of Rhus tox including its crude form have been evaluated for their effects on immune response in the in vivo and in vitro experimental models. METHODS: Rhus tox in the form of mother tincture, 6cH, 30cH, 200cH and 1000cH dilutions was tested through in vivo models including sheep red blood cells (SRBCs) induced cellular and humoral immune response in C57/BL6 mice. The effects of Rhus tox dilutions were also evaluated in vitro on the functions of human polymorphonuclear (PMN) cells such as phagocytosis and intracellular killing of Candida albicans, chemotaxis, and reduction of nitroblue tetrazolium (NBT) dye. RESULTS: Rhus tox was found to intensify SRBCs induced antibody titer and delayed type hypersensitivity response in mice. Even higher dilutions such as 200cH and 1000cH were found to affect the immune response; however, the crude form, mother tincture, 6cH and 30cH dilutions revealed more potent effects than the 200cH and 1000cH dilutions. In in vitro assays, all the dilutions exerted stimulation of phagocytosis, candidacidal activity and chemotaxis of human PMN cells. The NBT dye reduction assay revealed that oxidative processes in the PMN cells are accelerated in the presence of Rhus tox. This study shows that Rhus tox possesses immunostimulatory activity in its crude form as well as in homeopathically diluted forms. These effects appeared to be concentration dependent as higher dilutions had less potent effects.

[Effects and mechanisms of shidi capsule on adjuvant arthritis model].

OBJECTIVE: To probe into the effects and mechanisms of Shidi capsule on adjuvant arthritis model (AA). METHOD: SD rats, maleness, were stochastically divided into the normal comparison, the model, the dexamethasone (0.45 mg kg(-1)), total glucosides of paeony (150 mg kg(-1)) and Shidi capsule (gruffs) high (5400 mg kg(-1)), middle (2700 mg kg(-1)) and low (1350 mg kg(-1)) dose groups. Except the normal comparison group, rats in the other groups were injected Freund's complete adjuvant reagent into the rat' right metapedes to establish the AA model. Drugs were given by intragastric administration. Then paw swelling, the content of prostaglandin E2 in inflamed paws, MDA and SOD in blood serum, the spleen and thoracic gland index, and histopathology change of malleolus joint were observed. RESULT: The Shidi high, middle and low dose groups could inhibit the primary and secondary swelling of the rats' inflamed metapedes in a dose-dependent manner. Compared with the model group, each group of Shidi capsule could increase SOD,decrease MDA and PGE2, advance the spleen, and thoracic gland index, and relive the histopathology change of malleolus joint. CONCLUSION: Shidi capsule could inhibit the inflammation of synovial membrane obviously, relieve the degree of injury of cell, and therefore have therapeutic efficacy to AA rats.

The homeopathic antiarthitic preparation Zeel comp. N: a review of molecular and clinical data.

Zeel comp. N (Zeel) is a homeopathic medication that has been widely used for many years for the treatment of arthritic disorders in a large number of countries worldwide. In recent years, a growing body of clinical and molecular evidence has been accumulating that shed light on the possible antiarthritic effects of this preparation. A number of studies report anti-inflammatory effects from Zeel. In vitro studies have indicated Zeel-mediated inhibition of the pathways involving the enzymes cyclooxygenase-1 and -2, and also the 5-lipoxygenase pathways, affecting levels of both eicosanoids and leukotrienes. Thus, Zeel may reduce the main two classes of molecules responsible for arthritic pain and inflammation. This review describes recent research on Zeel and discusses the need for further studies to clarify the role of the compound in the antiarthritic armamentarium of complementary medicine.

[Study on mechanism underlying the treatment of rheumatoid arthritis by Keshiling].

OBJECTIVE: To explore the mechanism underlying the treatment of rheumatoid arthritis by Keshiling (KSL) in the rats model of FCA-induced arthritis. METHOD: The experimental arthritis was induced by FCA in the rats. The content of PGE2 in the inflammatory swelling toes was evaluated by ultraviolet spectrophotometric method. The ConA and LPS-induced lymphocytes proliferation and the production of interleukin-2 (IL-2) secreted by thymus were determined by MTT assay. RESULT: Results showed that the increases of lymphocyte proliferation and IL-2 production in AIA rats could be inhibited by KSL at the concentrations of 540 and 270 mg x kg(-1) in vivo and vitro. KSL at the same doses decreased the contents of PGE2 in inflammatory swelling toes, and the decreased A values were 25.6,16.1, 10.0 (A x 10(3)), respectively. After administration of KSL in vivo at 540 and 270 mg x kg(-1) the T lymphocyte proliferation were attenuated by 32.1% and 31.0%, and the production of IL-2 was inhibited by 17.5% and 14.0% respectively. While the inhibitory rates of T lymphocyte proliferation were reduced by 39.0% and 22.1% and the production of IL-2 was diminished by 27.3% and 18.2% respectively following the administration of KSL in vitro. CONCLUSION: KSL possesses the anti-inflammation function.

Safety and anti-inflammatory activity of curcumin: a component of tumeric (Curcuma longa).

INTRODUCTION: Tumeric is a spice that comes from the root Curcuma longa, a member of the ginger family, Zingaberaceae. In Ayurveda (Indian traditional medicine), tumeric has been used for its medicinal properties for various indications and through different routes of administration, including topically, orally, and by inhalation. Curcuminoids are components of tumeric, which include mainly curcumin (diferuloyl methane), demethoxycurcumin, and bisdemethoxycurcmin. OBJECTIVES: The goal of this systematic review of the literature was to summarize the literature on the safety and anti-inflammatory activity of curcumin. METHODS: A search of the computerized database MEDLINE (1966 to January 2002), a manual search of bibliographies of papers identified through MEDLINE, and an Internet search using multiple search engines for references on this topic was conducted. The PDR for Herbal Medicines, and four textbooks on herbal medicine and their bibliographies were also searched. RESULTS: A large number of studies on curcumin were identified. These included studies on the antioxidant, anti-inflammatory, antiviral, and antifungal properties of curcuminoids. Studies on the toxicity and anti-inflammatory properties of curcumin have included in vitro, animal, and human studies. A phase 1 human trial with 25 subjects using up to 8000 mg of curcumin per day for 3 months found no toxicity from curcumin. Five other human trials using 1125-2500 mg of curcumin per day have also found it to be safe. These human studies have found some evidence of anti-inflammatory activity of curcumin. The laboratory studies have identified a number of different molecules involved in inflammation that are inhibited by curcumin including phospholipase, lipooxygenase, cyclooxygenase 2, leukotrienes, thromboxane, prostaglandins, nitric oxide, collagenase, elastase, hyaluronidase, monocyte chemoattractant protein-1 (MCP-1), interferon-inducible protein, tumor necrosis factor (TNF), and interleukin-12 (IL-12). CONCLUSIONS: Curcumin has been demonstrated to be safe in six human trials and has demonstrated anti-inflammatory activity. It may exert its anti-inflammatory activity by inhibition of a number of different molecules that play a role in inflammation.

Comparison of the postoperative analgesic and sedative effects of carprofen and papaveretum in the dog.

Forty dogs undergoing a variety of surgical procedures were assigned randomly to one of two groups. All the animals were premedicated with acepromazine (0.05 mg/kg bodyweight) intramuscularly, and anaesthesia was induced with thiopentone sodium, or propofol in the case of lean animals, and maintained with halothane in an oxygen/nitrous oxide mixture using a non-rebreathing circuit. The dogs in group 1 were given papaveretum (0.2 mg/kg) slowly intravenously within 35 minutes of induction of anaesthesia and the dogs in group 2 were given carprofen (4 mg/kg) in the same way. The dogs were scored for sedation and pain by a trained theatre nurse, who did not know which group they belonged to, using a visual analogue scale, at 15, 30, 60, 120, 240 and 360 minutes after the halothane was switched off at the end of the procedure. Nine of the dogs were withdrawn from the trial (eight of them from the papaveretum group) because of inadequate pain relief and these animals were given pethidine (3 mg/kg intramuscularly) which produced adequate analgesia within 15 minutes in all but one case. Carprofen provided profound analgesia which was as effective and of longer duration than that produced by papaveretum, and was associated with significantly less postoperative sedation and a quicker return to the normal conscious state.

[Pharmacological studies on the comparison between the drainage and certified Fel Ursi].

A comparison between the drainage and certified Fel Ursi was studied by pharmacological methods. The results demonstrated that the drainage Fel Ursi was similar to the certified products in anti-inflammatory, antipyretic, sedative, anticonvulsive, antispasmodic and bacteriostatic actions.

[Effects of musk glucoprotein on the function of rat polymorphonuclear leukocytes activated by IL-8 in vitro].

OBJECTIVE: To investigate the effects of musk-1, a glucoprotein component isolated from the water extract of musk, on some functions of rat polymorphonuclear leukocytes activated by IL-8 in vitro. METHOD: An in vitro incubation system was used. Superoxide anion production was determined by cytochrome C reduction. beta-glucuronidase and lysozyme release was quantitated by enzyme reactions in which phenolph-thaleinglucuronic acid and Micrococcus Lysodeikticus were as the substrates, respectively. RESULTS: In comparison with control, musk-1 at concentration 1-100 micrograms.ml-1 can increase superoxide anion production by 91.7%-291%, and decrease beta-glucuronidase and lysozyme release by 2.2%-58.1% and 3.9%-39.8%, respectively. CONCLUSION: Inhibition of lysosomel enzyme release might be considered as one of mechanisms of antiinflammatory action of musk.

[Ancient and modern studies on medicinal ants].

Textual study of historical and modern scientific studies on chemical constituents and pharmacological actions of medicinal ants were carried out. The records in the ancient literatures conform to modern scientific research. Rational utilization and development were suggested.

[Effects of the glucoprotein component of musk on functions of rat polymorphonuclear leukocytes activated by LTB4 in vitro].

To investigate the effects of musk-1, a glucoprotein component isolated from the water extract of musk, on some functions of rat polymorphonuclear leukocytes activated by LTB4, an in vitro incubation system with rat polymorphonuclear leukocytes was used. The superoxide anion production was determined by cytochrome C reduction, and the beta-glucuronidase and lysozyme release was quantitated by enzyme reactions in which phenolphthaleinglucuronic acid and micrococcus lysodeikticus were used as the substrates. In comparison with the control, musk-1 at final concentrations of 1 microgram/ml-100 micrograms/ml can increase the superoxide anion production by 28.7%-202.1% and decrease the beta-glucuronidase and lysozyme release by 3%-46% and 6%-32% respectively in rat polymorphonuclear leukocytes. It is concluded that musk-1 can significantly affect the functions of rat polymorphonuclear leukocytes activated by LTB4. One of the mechanisms of this anti-inflammatory action of musk may consist in the inhibition of lysosomal enzyme release.

[Effects of musk glucoprotein on chemotaxis of polymorphonuclear leukocytes in vivo and in vitro].

OBJECTIVE: To investigate the effects of Musk glucoprotein on chemotaxis of Polymorphonuclear leukocytes(PMN). METHOD: The chemotaxis of PMN in abdominal cavity in rat induced by carboxymethyl cellulose(CMC) was used as an in vivo animal model and in in vitro it was evaluated by Boyden chamber. The concentration of cytosolic free Ca2+ was quantitated with the fluorescent Ca2+ indicator Fura-2. RESULT: The water extract of Musk at dose of 5, 20, 80 mg.kg-1 (s.c.) significantly inhibited the chemotaxis of PMN in rat; Musk-1 at concentration of 1-100 micrograms.mL-1 can significantly inhibit the chemotaxis of rabbit PMN in vitro; Musk-1 at concentration of 1-100 micrograms.mL-1 can significantly inhibit the increase of cytosolic Ca2+ concentration in PMN of rat. CONCLUSION: Part of mechanisms underlying antiinflammatory action of Musk is to inhibit the chemotaxis of PMN.

[Studies on mechanism of antiasthmatic and anti-inflammatory actions of CDCA].

OBJECTIVE: To observe antiasthmatic and anti-inflammatory actions mechanisms of CDCA. METHOD: The content of NO was determined by method of nitroreductase chromatometry in serum and trachea tissue. The content of cAMP was analysed by method of competitive protein-binding assay. The content of PGE2 was determined by method of ultraviolet spectrophotometry. RESULT: CDCA significantly decreased the content of NO of serum and trachea tissue in mice. CDCA increased greatly the content of cAMP of trachea tissue in rats. CDCA significantly decreased the content of PGE2 of trachea and lung tissue in mice. CONCLUSION: Mechanisms of antiasthmatic and anti-inflammatory actions of CDCA are related to increasing the content of cAMP in trachea tissue and decreasing the constituent of NO and PGE2 in body.

[Anti-inflammatory and analgesic effects of scolopendra from different habitats].

The water extracts of several species of the Chinese drug scolopendra from different habitats can inhibit obviously the increased permeability of abdominal blood capillaries and ear inflammation in mice. They can also raise the pain thresholds in mice during hot-plate and writhing tests. Their toxicity is very low.

[Pharmacodynamics study on "tao hua zhi xie granule"].

The anthraco-propulsion and the drug induced mice diarrhea models were used to observe the intestine propulsion and the anti-diarrhea effect of "Tao Hua Zhi Xie Granule" (THZXG), and the anti-inflammation effect of THZXG was studied. The results showed that THZXG could obviously reduce the incidence and frequency of the mice diarrhea induced by Folium Sennae and castor-oil, and propelling movement of mice small intestine after hypodermic injection of neostigmine. The actions were acted in a dose-dependent manner. 11.70 and 17.55 mg/kg THZXG(ig) could also inhibit the increased permeability of intraperitoneal capillary induced by acetic acid in mice.