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Αλέξανδρος Γ. Σφακιανάκης

Thursday, June 13, 2019

Ethnopharmacology

Antidiabetic and hypolipidemic efficacy of skin and seed extracts of Momordica cymbalaria on alloxan induced diabetic model in rats

Publication date: 15 September 2019

Source: Journal of Ethnopharmacology, Volume 241

Author(s): Abbirami Elangovan, Abinaya Subramanian, Siva Durairaj, Jeyadevi Ramachandran, Dinesh Kumar Lakshmanan, Guna Ravichandran, Gayathri Nambirajan, Sivasudha Thilagar

Abstract
Ethnopharmacological relevance

Momordica cymbalaria, a wild vegetable belongs to the Cucurbitaceae family, has long been used as a food and a remedy for diabetes mellitus in the Asian native medicinal system.

Aim of the study

This study aims to evaluate the efficacy of ethanolic extract of skin (EESK) and methanolic extract of seed (MESE) of M. cymbalaria (MC), for their hypoglycemic and hypolipidemic effects in alloxan induced diabetic rats.

Materials and methods

The diabetes induced rats were given skin and seed extracts at doses 250 and 500 mg/kg b.w. p.o. for 28 days. Alloxan monohydrate (120 mg/kg) was used to induce diabetes mellitus. Daily food and water intake were assessed. Blood glucose levels and body weights were measured every 7 days throughout the experiment. Antioxidant assays, different biochemical and glycemic parameters were evaluated. Histopathological studies on pancreas, liver and kidney were also studied.

Results

Treatment of EESK and MESE showed dose significant decrease in fasting blood glucose level (FBG) in experimental diabetic animals with significant reduction in food and water intake and increase in body weight. Findings confirmed the hypoglycemic and hypolipidemic effects of EESK and MESE in the experimental groups. The impaired glucose tolerance and altered activities of the hepatic enzymes such as AST, ALT and ALP levels of diabetic rats were significantly improved by the administration of EESK and MESE. Oral treatment with MC extract for 28 days demonstrated significant protective effects on the lipid profile, biochemical parameters and antioxidant levels. Besides, biochemical findings were supported by histopathological investigations.

Conclusion

These results suggest that the treatment with EESK and MESE of MC at a dose of 500 mg/kg b.w. have better protective effects against hyperglycemia, hyperlipidemia and oxidative stress generated during diabetes justifying the use of the plant in traditional systems of medicine.

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Neoline is the active ingredient of processed aconite root against murine peripheral neuropathic pain model, and its pharmacokinetics in rats

Publication date: 15 September 2019

Source: Journal of Ethnopharmacology, Volume 241

Author(s): Yohei Tanimura, Masato Yoshida, Kan'ichiro Ishiuchi, Masahiro Ohsawa, Toshiaki Makino

Abstract
Ethnopharmacological relevance

Processed aconite root (PA), the root of Aconitum carmichaeli (Ranunculaceae), is a crude drug used in traditional Chinese or Japanese kampo medicine to treat pain associated with coldness. In our previous study, PA and its active ingredient, neoline, alleviated oxaliplatin-induced peripheral neuropathy in mice.

Aim of the study

The present study investigated the effects of PA on a murine peripheral neuropathy model induced by intraperitoneal injection of paclitaxel and partial ligation of the sciatic nerve (Seltzer model), and identified its active ingredients.

Materials and methods

PA powder (1 g/kg/day) was orally administered, and either neoline or benzoylmesaconine (10 mg/kg/day) was subcutaneously injected into the murine model. Mechanical hyperalgesia was evaluated via the von Frey filament method. PA extract was orally administered to rats; blood samples were chronologically collected, and the plasma concentrations of Aconitum alkaloids were measured. The contents of Aconitum alkaloids in commercial PA products were also measured.

Results

PA extract and neoline significantly attenuated the mechanical hyperalgesia induced by either paclitaxel or partial ligation of the sciatic nerve in mice. In the plasma samples of rats treated with PA extract, higher concentrations of benzoylmesaconine and neoline were apparent among Aconitum alkaloids. The contents of benzoylmesaconine and neoline varied among PA products with different processing procedures. Subcutaneous injection of benzoylmesaconine did not attenuate the hyperalgesia induced by each paclitaxel, partial ligation of the sciatic nerve, or oxaliplatin in mice.

Conclusions

The present results indicate that PA and its active ingredient, neoline, are promising agents for the alleviation of neuropathic pain. Neoline can be used as a marker compound to determine the quality of the PA products for the treatment of neuropathic pain.

Graphical abstract

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Ethnobotanical survey of medicinal plants in central Abyan governorate, Yemen

Publication date: 15 September 2019

Source: Journal of Ethnopharmacology, Volume 241

Author(s): Mohamed Al-Fatimi

Abstract
Ethnopharmacological relevance

Traditional medicinal plant knowledge in Yemen is still only passed on orally from one generation to another. A few studies, mostly very limited in scope, have been dedicated to this indigenous knowledge so far. This paper presents the results of the first extensive field study in southern mainland Yemen, undertaken in three communities in the central region of Abyan governorate. The study is aimed to preserve the heritage of this indigenous knowledge and to explore and select local medicinal plants that promise high pharmacological efficacy for further pharmacological and phytochemical investigations.

Methods

356 indigenous informants (273 males, 83 females) were interviewed about the modes of application and uses of the medicinal plants in the region. The ethnobotanical data were recorded by semi-structured face-to-face interviews, substantiated by specimen collections and taxonomic identifications and quantitative data analysis including informant consensus factor (ICF) and number of use-reports.

Results

In total, 195 medicinal plant species (170 wild and 25 cultivated) belonging to 138 genera and 55 flowering plant families, were recorded for the treatment of 155 different ailments classified into 16 categories. Apocynaceae (25 species), Fabaceae (18 species), Euphorbiaceae (16 species) and Asteraceae (14 species) were the most frequently used plant families. Paste (86) followed by unprepared plant exudates (77) and decoction (55) were the most common herbal preparation modes. Dermal application (51.8%) was the most common administration route. The highest number of use reports and the maximum number of used medicinal plants were recorded for treatment of skin and gastrointestinal ailments. Forty-five species have never been reported in the ethnomedicinal literature before. Plant species with the most citations were Aloe vacillans Forssk. (malaria), Solanum incanum L. (tooth decay), Caralluma awdeliana (Deflers) A. Berger (diabetes), Tribulus terrestris L. (kidney stones), Aristolochia bracteolata Lam. (snake poison), Hydnora abyssinica A.Br. (stomach ulcer), Indigofera oblongifolia Forssk. (urine retention) and Chrozophora oblongifolia (Delile) A. Juss ex Spreng (haemorrhoids).

Conclusions

Significant traditional knowledge of the uses of local medicinal plant species was recorded for the first time for central Abyan in particular and southern Yemen in general. The study shows that the medicinal plants still play an important role in the primary health care in the study area. The ethnobotanical results provide a basis for further pharmacological, biological, pharmacognostical, and phytochemical investigations.

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A lipid-soluble extract of Pinellia pedatisecta Schott enhances antitumor T cell responses by restoring tumor-associated dendritic cell activation and maturation

Publication date: 15 September 2019

Source: Journal of Ethnopharmacology, Volume 241

Author(s): Yumeng Wang, Haixia Huang, Sheng Yao, Guiling Li, Congjian Xu, Yang Ye, Suiqi Gui

Abstract
Ethnopharmacological relevance

Pinellia pedatisecta Schott (PPS)is a traditional Chinese medicine functioning as reducing swelling and drying dampness. Pinellia pedatisecta Schott extract (PE) has been confirmed to suppress cervical tumor growth and modulate the antitumor CD4+T helper immunity towards Th1.

Aims

To explore the roles of PE in modulating tumor-associated dendritic cell (TADC) activation and function.

Methods

For in vivo studies, HPV+TC-1 mouse tumor models were conducted and treated with PE for 3 weeks (10 mg/kg/d or 20 mg/kg/day). The immune profiles of spleen, tumor-draining lymph nodes (TDLNs), tumor and serum were analyzed by flow cytometry and multiplexed bead-based immunoassay. For in vitro studies, TADCs were generated by tumor-conditioned medium and treated with PE solution. The maturation and function of TADCs were evaluated by flow cytometry, ELISA, mixed lymphocyte reaction (MLR) and cytotoxic T lymphocyte (CTL) assay. Furthermore, the effect of PE on SOCS1 pathway was examined by western blotting and real time PCR.

Results

PE upregulated the expression of major histocompatibility complex class II (MHCII) and costimulatory molecules CD80 and CD86 on TADCs and promoted IL-12 secretion from TADCs. In addition, PE-treated TADCs promoted the proliferation of CD4+ and CD8+ T cells and induced the differentiation of IFN-γ+CD4+ and GZMB+CD8+ T cells. PE-treated TADCs also elicited a more powerful antigen-specific cytotoxic T lymphocyte (CTL) response. Furthermore, PE treatment in vivo enhanced the proliferation, activated the functional ability (increased Ki67, CD137, GZMB or IFN-γ, TNF-α expression) and reversed the exhaustion (impaired CD95 or PD-1 expression) of antitumor T cells. Mechanistically, PE inhibited SOCS1-restrained JAK2 activation in TADCs.

Conclusions

PE efficiently restored the immature status of TADCs and enhanced their function as antigen-presenting cells to further elicit antitumor Th1 and CTL responses, suggesting that PE may be a potential immunomodulatory drug for cancer treatment.

Graphical abstract

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Binding of the polysaccharide from Acanthopanax giraldii Harms to toll-like receptor 4 activates macrophages

Publication date: 15 September 2019

Source: Journal of Ethnopharmacology, Volume 241

Author(s): Qingqing Li, Zhiting Chen, Zhilu Xu, Shaoyun Han, Huihui Hao, Jiang Wu, Fengxiang Sun, Xiaoyan Fu, Ruyue Li, Birong Zheng, Xiaoxiao Guo, Tongtong Zhang, Yong Chen

Abstract
Ethnopharmacological relevance

The traditional Chinese medicine, Acanthopanax giraldii Harms, is commonly used to treat arthralgia due to wind, cold and dampness, as well as weakness in the feet and knees. Its other reported effects include eliminating flatulence, strengthening muscles and bones, and delaying aging. The polysaccharides in A. giraldii Harms are the major bioactive substances that confer the herb's antioxidant properties as well as anticancer and antiviral effects.

Aims of the study

To elucidate the underlying mechanism and signaling cascade involved in the homogeneous A. giraldii Harms polysaccharide II (AHP–II)–mediated immunomodulation of mice macrophages.

Materials and methods

The phagocytosis of neutral red and the production of nitric oxide, interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α), were measured to determine AHP–II–induced macrophage activation. Confocal microscopy and flow cytometry were used to confirm the binding of AHP-II to macrophages. The involvement of Toll-like receptor (TLR) 4 in AHP–II–induced macrophage activation was demonstrated using antibody blocking and macrophages from C3H/HeJ TLR4-mutant mice. Western blotting was used to map AHP–II–induced downstream signaling pathways.

Results

AHP-II increased the phagocytosis of macrophages and the release of nitric oxide, IL-6 and TNF-α cytokines. Direct, saturable and reversible binding of AHP-II to macrophages was observed, while it can be inhibited by the anti-TLR4 antibody. In addition, the presence of the anti-TLR4 antibody inhibited AHP–II–induced macrophage IL-6 and TNF-α production in the peritoneal macrophages of C3H/HeJ mice. Moreover, AHP–II–TLR4-stimulated macrophages activate the downstream intracellular ERK and JNK/nuclear factor (NF)-κB signaling pathways. In addition, the AHP–II–mediated regulation of IL-6 and TNF-α production from macrophages was greatly affected by specific ERK, JNK and NF-κB inhibitors.

Conclusion

Our study elucidated the immunomodulatory mechanism of AHP-II in macrophage activation and identified TLR4 as the main receptor coordinating AHP-II binding. Our findings suggest AHP-II may be used as a novel immunopotentiator for medical purposes.

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Utilization trends in traditional Chinese medicine for acute myocardial infarction

Publication date: 15 September 2019

Source: Journal of Ethnopharmacology, Volume 241

Author(s): Chung-Yen Lu, Pei-Chin Lu, Pei-Chun Chen

Abstract
Ethnopharmacological relevance

Among heart diseases, acute myocardial infarction (AMI) is the most serious and life-threatening emergency. In Taiwan, heart disease has consistently ranked second among the top 10 leading causes of death since 2007, second only to malignant tumors; however, population-based studies on the use of traditional Chinese medicine (TCM) in AMI cases are limited.

Aim of the study: This study investigated the characteristics of TCM users and prescriptions of TCM, and their differences between two cohorts of patients with AMI, identified 10 years apart.

Materials and methods

A cross-sectional study was conducted using the Taiwan National Health Insurance claims database. From among two random sample of 1 million beneficiaries selected from the claims database, we identified two cohorts of patients with first hospitalization for AMI in between 2000–2001 and 2010–2011. Patients who had received TCM therapy within one year after hospital discharge were defined as TCM users, whereas, all the other patients with AMI were considered non-users of TCM. We compared the characteristics of TCM use and the patterns of prescriptions between the two cohorts.

Results

The proportion of patients receiving TCM care was similar between the two AMI cohorts; 20% (85/418) of the patients were diagnosed in 2000–2001 and 21% (169/817) in 2010–2011. In the 2010–2011 AMI cohort, the proportion of men was smaller among TCM users than non-users, and TCM users were less likely to have hyperlipidemia. Among TCM users, the most frequently prescribed herb was Dan-shen (Salvia miltiorrhiza Bunge, Salvia root) in both cohorts. The most commonly used Chinese herbal formulations were Xue-Fu-Zhu-Yu-Tang (Blood Mansion Dispel Stasis) for the 2000–2001 cohort and Zhi-Gan-Cao-Tang (Honey-Fried Licorice Decoction) for the 2010–2011 cohort.

Conclusions

This study revealed the differences in the prescription frequency of Chinese herbal formulation among the two cohort of patients with AMI, suggesting that the practice of prescribing TCM has evolved from post-antique formula to classical remedies during the 10 years evaluated. Further investigations are needed to evaluate if the change in the utilization of Chinese herbal formulations impacts the effectiveness of the treatment.

Graphical abstract

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Anxiolytic- and anxiogenic-like effects of Montanoa tomentosa (Asteraceae): Dependence on the endocrine condition

Publication date: 15 September 2019

Source: Journal of Ethnopharmacology, Volume 241

Author(s): Erika Estrada-Camarena, Isabel Sollozo-Dupont, Dannia Islas-Preciado, María Eva González-Trujano, Miguel Carro-Juárez, Carolina López-Rubalcava

Abstract
Ethnopharmacological relevance

Montanoa tomentosa Cerv. (MT) is a native plant from Mexico used in traditional medicine as a remedy for reproductive impairments and relaxing effects. In previous studies, it has been shown that the endocrine state could modify the antianxiety-like actions of anxiolytic compounds. Although women are the primary user of MT, no studies have evaluated the potential impact of the endocrine milieu on its anti-anxiety actions.

Aims of the study:Ascertain the antianxiety effects of M. tomentosa in rats with different hormonal conditions, and to analyze the participation of the GABAA receptor in ovariectomized rats treated with MT.

Materials and methods

The animal model of anxiety used was the elevated plus-maze (EPM). Rats' endocrine conditions were: a) Low hormone levels (rats in diestrus I and II phases); b) High hormone levels (proestrus/estrus phases); c) No hormones (ovariectomized rats); and d) Rats under progesterone withdrawal (PW). To evaluate the participation of the GABAA receptor in the anxiolytic-like action of MT the antagonist picrotoxin was used.

Results

Results showed that MT induced dose-dependent anxiolytic-like actions in rats with low hormone level conditions. Also, MT reduced anxiety-like behavior in female rats under PW, in contrast to diazepam which was ineffective. MT's anxiolytic-like effect was blocked by picrotoxin, suggesting the participation of the GABAA receptor complex. However, increased anxiety-like behavior was observed in rats with a high hormone level condition and low doses of MT.

Conclusions

Beneficial anxiolytic-like actions of MT are observed under low hormone conditions, particularly in the PW challenge (a condition that can be related to a premenstrual period). Furthermore, the participation of the GABAAreceptor is evidenced. However, hormonal variations could induce the opposite effects, hence women should be cautious.

Graphical abstract

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Hazard assessment of Maerua subcordata (Gilg) DeWolf. for selected endpoints using a battery of in vitro tests

Publication date: 15 September 2019

Source: Journal of Ethnopharmacology, Volume 241

Author(s): Mebrahtom Gebrelibanos Hiben, Lenny Kamelia, Laura de Haan, Bert Spenkelink, Sebastiaan Wesseling, Jacques Vervoort, Ivonne M.C.M. Rietjens

Abstract
Ethnopharmacological relevance

Maerua subcordata (Gilg) DeWolf is a medicinal and wild food plant growing mainly in east Africa. Especially its root tuber is widely used in traditional medicine to treat several infectious and chronic diseases but also in some toxicity implications like use as abortifacient.

Aim of the study

the present study applied in silico and in vitro tests to identify possible hazards of M. subcordata (fruit, leaf, root, seed) methanol extracts focussing on developmental toxicity.

Materials and methods

Ames test, estrogen receptor alpha (ERα) assay, aryl hydrocarbon receptor (AhR) assay, embryonic stem cell test (EST), and zebrafish embryotoxicity test (ZET) were employed. Besides, a Derek Nexus toxicity prediction was performed on candidate structures obtained from metabolomics profiling of the extracts using liquid chromatography coupled to multistage mass spectroscopy (LC/MSn) and a MAGMa software based structural annotation.

Results

Glucosinolates, which degrade to isothiocyanates, and biogenic amines were among the candidate molecules identified in the extracts by LC/MSn - MAGMa software structural annotation. Isothiocyanates and some other candidate molecules suggested a positive mutagenicity alert in Derek toxicity predictions. All the extracts showed negative mutagenicity in the Ames test. However, the Derek predictions also identified endocrine and developmental toxicity as possible endpoints of concern. This was further assessed using in vitro tests. Results obtained reveal that leaf extract shows AhR and ERα agonist activities, inhibited differentiation of ES-D3 stem cells into contracting cardiomyocytes in the EST (p < 0.001) as well as inhibited hatching (p < 0.01) and showed acute toxicity (p < 0.01) in the ZET. Also, the fruit extract showed toxicity (p < 0.05) towards zebrafish embryos and both fruit and seed extracts showed AhR agonist activities while root extract was devoid of activity in all in vitro assays.

Conclusion

The leaf extract tests positive in in vitro tests that may point towards a developmental toxicity hazard. The current evaluations did not raise concerns of genotoxicity or developmental toxicity for the fruit, seed and root extracts. This is important given the use of especially these parts of M. subcordata, in traditional medicine and/or as (famine) food.

Graphical abstract

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The effect of isoflavaspidic acid PB extracted from Dryopteris fragrans (L.) Schott on planktonic and biofilm growth of dermatophytes and the possible mechanism of antibiofilm

Publication date: 15 September 2019

Source: Journal of Ethnopharmacology, Volume 241

Author(s): Haoqi Lin, Xueping Liu, Zhibin Shen, Wanqiu Cheng, Zhijun Zeng, Yanfen Chen, Chunping Tang, Tao Jiang

Abstract
Ethnopharmacological relevance

Dryopteris fragrans (L.) Schott (D. fragrans), a deciduous perennial herb, has been traditionally used for treatment of various skin diseases in Heilongjiang province of China for many years. Phloroglucinol derivatives extracted from D. fragrans were the most effective fraction against dermatophytes. Isoflavaspidic acid PB is a typically phloroglucinol derivative which extracted from D. fragrans and has been reported to exert anti-fungal activities against several dermatophytes.

Aim of the study. This study aimed to evaluate anti-fungal and anti-biofilm activity of isoflavaspidic acid PB on planktonic and biofilm growth of dermatophytes and explore possible mechanisms of anti-biofilm.

Materials and methods

Minimal inhibitory concentrations (MIC) and minimal fungicidal concentrations (MFC) of isoflavaspidic acid PB against 25 isolates of dermatophytes were determined by the Clinical and Laboratory Standards Institute (CLSI) M38-A2 method. The effects of isoflavaspidic acid PB on dermatophytes biofilm formation and pre-formed biofilm were assessed by 2.3-bis (2-methoxy-4-nitro-5-sulfophenyl)-5-[carbonyl (phenylamino)]-2H-tetrazolium hydroxide (XTT) assay. Morphology of mature biofilm were observed by Scanning Electron Microscope (SEM). Biomass, exopolysaccharide and ergosterol content of mature biofilm were analyzed by gravimetric analysis, anthranone sulfuric acid method and Ultra Performance Liquid Chromatography (UPLC) assay respectively.

Result

The MIC and MFC ranges of isoflavaspidic acid PB against 25 isolates of dermatophytes were 20–80 μg/mL and 40–80 μg/mL respectively. Isoflavaspidic acid PB (2 MIC) inhibited not only Trichophyton biofilm formation (54.8% ∼ 81.2%) but also the metabolic activity of mature biofilm (20.7% ∼ 44.2%). The result of SEM showed that isoflavaspidic acid PB (8 MIC) could destroy the morphology of hyphae seriously. Comparing with control group, biomass, exopolysaccharide and ergosterol content of the mature biofilm under isoflavaspidic acid PB (8 MIC) were significantly decreased (P < 0.01).

Conclusion

Isoflavaspidic acid PB had anti-fungal and fungicidal activities against dermatophytes. Isoflavaspidic acid PB could inhibit the biofilm of Trichophyton. The mechanism might be related to the decline of the biofilm biomass, exopolysaccharide and ergosterol content. These results showed that isoflavaspidic acid PB could be explored for promising anti-biofilm drugs.

Graphical abstract

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Amelioration of dry eye syndrome in db/db mice with diabetes mellitus by treatment with Tibetan Medicine Formula Jikan Mingmu Drops

Publication date: 15 September 2019

Source: Journal of Ethnopharmacology, Volume 241

Author(s): Xiaopeng Ai, Ya Hou, Xiaobo Wang, Xiaoyan Wang, Yusheng Liang, Zhengwen Zhu, Ping Wang, Yong Zeng, Xianjia Li, Xianrong Lai, Xianli Meng, Qi'en Li

Abstract
Ethnopharmacological relevance

Jikan Mingmu Drops (JMD), a traditional Tibetan medicine containing six herbs, has been used to treat dry eye syndrome (DES) in individuals with diabetes mellitus.

Aim of study

However, the activity of JMD ameliorates DES with diabetes mellitus has not been previously examined. The aim of the study is to investigate the molecular mechanism of JMD on db/db mice.

Materials and methods

The main chemical constituents of JMD were analyzed by high-performance liquid chromatography and gas chromatography–mass spectrometry. DES was then induced in db/db mice by applying 0.2% benzalkonium chloride to the ocular surface for 7 days. Eye drops containing JMD (0.25, 0.5, or 1 g/mL) or vehicle subsequently were administered three times daily for another 7 days, and the therapeutic effects were evaluated by phenol red thread tear and sodium fluorescein tests. Conjunctival specimens were subjected to hematoxylin and eosin staining and periodic acid–Schiff staining to examine pathological changes and number of goblet cells. ELISA was performed to assess the levels of various inflammatory cytokines.

Results

JMD contains hydroxysafflor yellow A, magnoflorine, jatrorrhizine hydrochloride, palmatine hydrochloride, berberine hydrochloride, gallic acid, ellagic acid, tauroursodeoxycholic acid, camphor, isoborneol, borneol, trans-cinnamic acid, and muscone. JMD treatment significantly increased the tear volume, decreased the corneal fluorescein staining score, restored the morphology and structure of conjunctival epithelial cells, and markedly downregulated the levels of interleukin (IL)-6, IL-17α, IL-1β, tumor necrosis factor-α, and vascular endothelial growth factor in the conjunctiva. Further data showed that these protective effects were accompanied by inhibition of inflammation in a dose-dependent manner.

Conclusions

Amelioration of DES in db/db mice with diabetes mellitus by treatment with Tibetan medicine formula JMD maybe related to its anti-inflammatory effects.

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Alexandros Sfakianakis
Anapafseos 5 . Agios Nikolaos
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