The in vivo and in vitro diabetic wound healing effects of a 2-herb formula and its mechanisms of action

Ethnopharmacological relevance

The herbs Radix Astragali (RA) and Radix Rehmanniae (RR) have long been used in traditional Chinese Medicine and serve as the principal herbs in treating diabetic foot ulcer.

Aim of study

Diabetic complications, such as foot ulcer, impose major public health burdens worldwide. In our previous clinical studies, two Chinese medicine formulae F1 and F2 have achieved over 80% limb salvage. A simplified 2-herb formula (NF3) comprising of RA and RR in the ratio of 2:1 was used for further study. NF3 was examined for the ulcer healing effect in diabetic rats, and its potential mechanisms of action in fibroblast proliferation, angiogenesis and anti-inflammation in vitro.

Materials and methods

A chemically induced diabetic foot ulcer rat model was used for studying the wound healing effect. In the in vitro mechanistic studies, human fibroblast cells (Hs27), human umbilical vein endothelial cells (HUVEC) and mouse macrophage cells (RAW264.7) were assessed for tissue regeneration, angiogenesis and anti-inflammatory activities, respectively.

Results

Our in vivo results demonstrated a significant reduction of wound area at day 8 in NF3 (0.98 g/kg) group as compared to control (p < 0.01). NF3 could significantly stimulate Hs27 proliferation in a dose dependent manner (p < 0.05). Besides, NF3 could significantly increase the cell migration and tube formation (p < 0.05-0.001) of HUVEC in the angiogenesis study. Furthermore, significant inhibition of nitric oxide production (p < 0.01) was found in NF3-treated macrophage cells, suggesting its anti-inflammatory activity.

Conclusions

Our study presents for the first time scientific evidence towards the efficacy of the two-herb formula NF3 in enhancing diabetic wound healing through the actions of tissue regeneration, angiogenesis and anti-inflammation.     

Effects of Chinese herbal cataplasm Xiaozhang Tie on cirrhotic ascites

Background

Conventional methods of treating cirrhotic ascites are inadequate. We sought to identify a novel, effective approach to relieve the suffering of patients with cirrhotic ascites.

Aim of the study

To investigate the efficacy of Xiaozhang Tie, a traditional Chinese herbal cataplasm composed of dahuang (Rheum palmatum L.), laifuzi (Raphanus sativus L.), concocted gansui (Euphorbia kansui T.N. Liou ex T.P. Wang), chenxiang [Aquilaria sinensis (Lour.) Gilg], dingxiang (Eugenia caryophyllata Thunb.), bingpian (Borneolum syntheticum) and shexiang (artificial Moschus), as an adjuvant in treating cirrhotic ascites.

Materials and methods

A multicenter, randomized, placebo-controlled trial was conducted. One hundred patients with cirrhotic ascites were divided into two groups of equal size. The test group took an umbilical compress with Xiaozhang Tie for 30 days while the control group was administered an umbilical compress with placebo, in addition to primary therapy. Efficacy was evaluated according to the criteria including ascites volume, urine 24-h volume, abdominal circumference, body weight, abdominal distention, appetite, flatus and defecation.

Results

Ninety-two patients completed the study, 7 were withdrawn and 1 was excluded. The effective rate of grades I and II was 63.3% for the test group (n = 49) and 38.0% for the control one (n = 50). Both groups showed decreased body weight and abdominal circumference, increased urine volume and improved symptoms after treatment. However, the differences between pre-treatment and post-treatment in body weight, abdominal circumference and urine volume were 8.7 ± 5.8 kg, 12.4 ± 8.3 cm and 683 ± 644 ml respectively in the test group, noticeably higher than those in the control group, which were 5.3 ± 4.6 kg, 8.0 ± 6.5 cm and 372 ± 697 ml, respectively. The ranking orders of the symptoms of the test group were significantly lower than those of the control group after treatment. No severe adverse reactions were seen.

Conclusion

Xiaozhang Tie as an adjuvant to primary therapy of cirrhotic ascites is safe and shows a remarkable efficacy on relieving abdominal distention.     

Optimized project of traditional Chinese medicine in treating chronic kidney disease stage 3: a multicenter double-blinded randomized controlled trial

Ethnopharmacological relevance

Stage 3 is the key phase of chronic kidney disease. Traditional Chinese medicine (TCM) has been used for the treatment of chronic kidney disease. But a large sample trial is desirable.

Materials and methods

A total of 578 Chinese patients with primary glomerulonephritis in CKD stage 3 were randomly assigned to three groups: patients received TCM (TCM group), benazepril (Ben group), TCM combined with benazepril (TCM + Ben group). Patients were followed up for 24 weeks. The primary endpoint was the time to the composite of 50% increased of serum creatinine, end stage renal disease or death.

Results

eGFR in the TCM and the TCM + Ben group were improved (week 24 vs. baseline, P < 0.05) while eGFR in the Ben group was decreased (week 24 vs. baseline, P > 0.05). 24 h urinary protein excretion (UP) and urinary albumin/creatinine (UAlb/Cr) were decreased in the TCM + Ben (week 24 vs. baseline, P < 0.05) and the Ben group (week 24 vs. baseline, P > 0.05). UP and UAlb/Cr were increased in the TCM group to week 12, then were stable (week 24 vs. baseline, P < 0.05). The hemoglobin in the TCM group was also improved (week 24 vs. baseline, P < 0.05). The accumulative survival rate in the TCM + Ben group was higher than that in the TCM group and the Ben group (P = 0.044). Side effects in the TCM group were the lowest in these groups (P < 0.05). The patients with dry cough in the TCM + Ben group and the Ben group were increased as compared with the TCM group (P < 0.05). Hyperkalemia happened less frequently in the TCM group as compared with the other two groups (P = 0.052).

Conclusions

For the patients with CKD stage 3, TCM can improve eGFR and hemoglobin with lower side effects. Benazepril significantly decreased the proteinuria. Chinese medicine integrated with benazepril can ameliorate renal function and decrease proteinuria synergistically.     

Zhen-wu-tang, a blended traditional Chinese herbal medicine, ameliorates proteinuria and renal damage of streptozotocin-induced diabetic nephropathy in rats

Ethnopharmacological relevance

Zhen-wu-tang (ZWT) is a blended traditional Chinese medicine specifically used for various kidney diseases.

Aim of the study

The present study is to evaluate the effects of ZWT on diabetic nephropathy and investigate the potential anti-diabetic mechanisms.

Materials and methods

Streptozotocin (STZ)-induced diabetic rats were orally administrated ZWT (80,320 mg/kg b.w.) once a day, for a period of 12 weeks. Body weight, urinary volume, urine protein and blood biochemical parameters were measured every 3 weeks. At the end of the observation period, kidneys were isolated for histology, angiotensin II content determination, real time PCR and Western blot analyses.

Results

ZWT (320 mg/kg b.w.) prevented body weight loss, reduced polyurea, urinary protein excretion, serum creatinine and blood urea nitrogen, although it did not alter the hyperglycemia. It ameliorated glomerular hypertrophy and expansion of the mesangial area, swell and effacement of foot process. It also suppressed the increased expression of angiotensin II, nephrin and podocin.

Conclusions

Data from this study suggest that ZWT possess a protective effect on renal damage of STZ-induced diabetic nephropathy in rats, by suppressing hyperactivity of renal renin-angiotensin system that turns in to modulate renal nephrin and podocin expressions, thereby protecting podocyte from injury.     

Effect of khat chewing on periodontal pathogens in subgingival biofilm from chronic periodontitis patients

Aims

Existing in vitro and in vivo data suggest that khat may have a favorable effect on periodontal microbiota. The purpose of this study was to assess the effect of khat chewing on major periodontal pathogens in subgingival plaque samples from subjects with chronic periodontitis.

Materials and methods

40 subgingival plaque samples were obtained from periodontitis and healthy sites of 10 khat chewers (40 y median age) and 10 khat non-chewers (37.5 y median age) with chronic periodontitis. Absolute and relative counts of 6 periodontal pathogens were determined in each sample using highly sensitive and specific Taqman real-time PCR assays. Data were analyzed using an ordinal regression model.

Results

Significantly more total bacteria were detected in samples from the periodontitis sites of the khat chewers (OR = 20). Treponema denticola was present at significantly higher absolute counts at the healthy as well as periodontitis sites of the khat chewers (OR = 3.13 and 13, respectively). However, the khat chewers harbored significantly lower absolute counts of Porphyromonas gingivalis at the healthy sites (OR = 0.07). Furthermore, khat chewing was significantly associated with lower relative counts of Porphyromonas gingivalis, fusobacterium ssp., prevotella ssp. and Parvimonas micra-like species in subgingival plaque samples from both healthy and periodontitis sites (OR = 0.11-0.33). Only Treponema denticola was found in higher relative counts at the healthy sites of the khat chewers (OR = 2.98).

Conclusions

Overall, there was a lower burden of pathogens in the khat chewers. Findings from the current study are suggestive of a potential prebiotic effect for khat on periodontal microbiota.     

The effect of bamboo extract on hepatic biotransforming enzymes - Findings from an obese-diabetic mouse model

Aim of the study

Bamboo leaves are used as a component in traditional Chinese medicine for the anti-inflammatory function. Our previous studies have demonstrated that an ethanol/water extract from Phyllostachys edulis ameliorated obesity-associated chronic systemic inflammation in mice, and therefore relieving the symptoms of type 2 diabetes. The aim of this project was to further investigate the effects of this bamboo extract on hepatic biotransformation enzymes in both lean and obese mice, as an initial step in the toxicological evaluation of using this traditional medicine in obese/diabetic population.

Materials and methods

Male C57BL/6J mice were randomized to 4 groups and fed standard (10% kcal from fat) diet with or without bamboo extract supplementation at a dose of 10 gram per kilogram diet (n = 10 and n = 9, respectively), or high fat (45% kcal from fat) diet with or without bamboo extract (n = 8 and N = 7, respectively). The dietary treatment lasted for 6 months. Subsequently, the activities and expression of the major Phase I and II hepatic biotransformation enzymes were assessed in subcellular fractions from murine livers.

Results

Three groups of mice, lean bamboo extract-supplemented, obese/diabetic, and bamboo extract-supplemented obese/diabetic, showed greater activities of cytochromes P450 1a2 and 3a11 compared to control but no changes in the expression level of these proteins. For Phase II enzymes, bamboo extract supplementation in lean mice caused decreased glutathione-S-transferase activity (−12%) and greater uridine diphosphate glucuronosyltransferase activity (+46%), but had no effect on sulfotransferase activity. Conversely, the obese/diabetic condition itself increased glutathione-S-transferase and uridine diphosphate glucuronosyltransferase activities, but decreased total sulfotransferase activity and sulfotransferase 2a1 expression.

Conclusions

Bamboo extract and obesity/diabetes show significant independent effects on hepatic biotransformation as well as interaction effects in mice. These changes may alter the clearance of endo- and xenobiotics, including bamboo extract itself, hence this effect should be carefully considered in the medicinal application of bamboo extract as it has potential to alter its own metabolism and that of other medications concurrently administered to obese diabetic patients.     

Comparative effects of Rauwolfia vomitoria and chlorpromazine on locomotor behaviour and anxiety in mice

Aim of the study

Since remedies for mental disorders have been sought through both orthodox and traditional medicine this study compared the effects of the antipsychotic, chlorpromazine (Cpz), the herb Rauwolfia vomitoria (RV) and its alkaloid reserpine (Res) in mice.

Materials and methods

Ninety male CD-1 strain of mice (75-80 days old; 30-34 g body weight) were divided into 3 major groups and each consisting 5 subgroups (n = 6). Cpz (0.0, 0.25, 1.0, 2.0 and 4.0 mg/kg, i.p.), was administered 30 min before testing. RV (0.0, 0.25, 1.0, 2.0 and 4.0 mg/kg, i.p.) and Res (0.0, 0.1, 0.4, 0.8, 1.6 mg/kg, i.p.) were administered 24 h before testing. The open field test was used to assess locomotor and exploratory behaviour, acceleratory rotarod for motor coordination, light/dark box for anxiety.

Results

CPZ dose-dependently decreased locomotor and exploration behaviour and impaired motor coordination (p < 0.01). RV also decreased locomotor behaviour at 4.0 mg/kg (p < 0.5) but did not alter exploration and motor coordination. Res however, decreased locomotion and exploration and impaired motor coordination 0.8 and 1.6 mg/kg (p < 0.05). In the light/dark box, CPZ increased anxiety related behaviour at 1.0, 2.0 mg/kg (p < 0.05) whereas RV dose-dependently decreased anxiety from 1.0 to 4.0 mg/kg (p < 0.01). Res, unlike RV, dose-dependently increased anxiety related behaviour from 0.4 to 1.6 mg/kg.

Conclusion

Root bark extract from Rauwolfia vomitoria produced better behavioural effects with less distortion in motor coordination when compared to chlorpromazine and so has a great potential as an alternative antipsychotic agent compared to chlorpromazine. Since Res did not produce same effects as RV, the effect of RV may not be due solely to Res as claimed.     

Ethno-veterinary uses and informants consensus factor of medicinal plants of Sariska region, Rajasthan, India

Aim of study

The study was conducted in Sariska region of Rajasthan, India to identify the important species used for ethno-veterinary medicine; finding out methods for various ethno-veterinary medicine preparations, and calculate the informant consensus factor (ICF) in relation to medicinal plant use.

Materials and methods

An ethno-veterinary survey was carried out in the Sariska region of Rajasthan, India. A total of 490 informants (287 men, 203 women) belonging mainly to families which had strong links with animal keeping activities of the area were interviewed using ‘specimen display’ method and forest walk method.

Results

The highest ICF (0.61) was scored for the digestive problems including ailments stomachache, indigestion, liver expansion, diarrhea, intestinal worms, and stomach disorder. Citrullus colocynthis is used for fever and general sickness, with a highest use value (UV) of 0.62. While Pedalium murex, and Ziziphus nummularia used for diarrhea (UV = 0.57) and Azadirachta indica, used as antiseptic, in foot and mouth disease and prevention from diseases were reported with a UV 0.51.

Conclusions

The documentation of this inherited rich traditional ethno-medicinal knowledge has provided novel information and this will not only provide recognition of this undocumented knowledge but will also help in conservation of such rare, gradually vanishing important ethno-veterinary species. It will also provide new pharmacological dimensions for better health care of the human being regarding many ailments.     

Polyphyllin D, a steroidal saponin from Paris polyphylla, inhibits endothelial cell functions in vitro and angiogenesis in zebrafish embryos in vivo

Ethnopharmacological relevance

Angiogenesis, the process of blood vessel formation, is critical to tumour growth. The importance of angiogenesis in tumour development has lead to the development of anti-angiogenic strategies to inhibit tumour growth. In this study, polyphyllin D (PD), an active component in Chinese herb, Paris polyphylla, was evaluated for its potential anti-angiogenic effects.

Materials and methods

The inhibitory effects of PD on three important processes involved in angiogenesis, i.e. proliferation, migration and differentiation were examined using human microvascular endothelial cell line HMEC-1 by MTT assay, scratch assay and tube formation assay, respectively. Using zebrafish embryos as an animal model of angiogenesis, the anti-angiogenic effect of PD was further verified in vivo.

Results

PD suppressed the growth of HMEC-1 cells at 0.1-0.4 μM without toxic effects. At 0.3 μM and 0.4 μM, PD significantly inhibited endothelial cell migration and capillary tube formation. About 70% of the zebrafish embryos showed defects in intersegmental vessel formation upon treatment with PD at concentrations of 0.156 μM and 0.313 μM.

Conclusion

The anti-angiogenic effects of PD have been explored in the study which implied a potential therapeutic development of PD in cancer treatment.     

Extracts of Arisaema rhizomatum C.E.C. Fischer attenuate inflammatory response on collagen-induced arthritis in BALB/c mice

Aim of the study

Arisaema rhizomatum C.E.C. Fischer (ARCF), called as “Xuelijian”, a local herb just growing in China, has been used as a traditional ethnic Chinese medicine for long because of its remarkable activity to alleviate pain and inflammation for patients suffering from rheumatism among the people with weak side-effect. However, rare study on the anti-arthritic activity of ARCF has been reported in vivo. The aim of this study is to investigate the protective effect of the herb on collagen-induced arthritis in mice and explore the potential immunological mechanisms.

Materials and methods

CIA was induced in male BALB/c mice by been subcutaneously injected type II bovine collagen (CII) for twice. The combined MeOH extract (ME) of ARCF rhizome was successively partitioned into four fractions with petroleum ether (PE), ethyl acetate (EE), n-butyl alcohol (n-BE) and water (WE). After the second collagen immunization, mice were administered orally with different doses of ME, EE and n-BE (ME 130, 261, 522 mg kg⁻¹; EE 10.2, 20.4, 40.8 mg kg⁻¹; n-BE 52, 104, 208 mg kg⁻¹) every other day for 3 weeks. The progression of edema of paws and knee joints was inspected by using a vernier calliper every 3 days from the 10th day after the first injection to the end of the experiment. The spleen index was measured and the knee joint destruction was observed by pathological sections. Levels of inflammatory cytokines including tumor necrosis factor alpha (TNF-α), interleukin-1 beta (IL-1β), interleukin-6 (IL-6), interleukin-33 (IL-33 or IL-1F11) and rheumatoid factor (RF) in serum were measured by ELISA.

Results

Administration of ME, EE and n-BE significantly suppressed paws and joints swelling and reduced the spleen indexes. Pathological examination demonstrated that ARCF effectively protected anklebone and cartilage from being eroded versus vehicle-treated mice. Moreover, the serum levels of inflammatory cytokines TNF-α, IL-1β, IL-6, IL-33 and RF were markedly lowered in ARCF treated groups compared with the control group (p < 0.05).

Conclusion

Our studies demonstrate that administration of ARCF is obviously suppressed the progression of CIA. The anti-arthritic effectiveness of ARCF will make the herb a strong candidate for further clinical trials on RA patients.     

Reproductive toxicity of Rhizoma Sparganii (Sparganium stoloniferum Buch.-Ham.) in mice: mechanisms of anti-angiogenesis and anti-estrogen pharmacologic activities

Ethnopharmacological relevance

Indications and preliminary studies of Rhizoma Sparganii (RS) suggest its pharmacological mechanism is involved with endocrine/angiogenesis functions. We therefore studied its potential toxicity on reproduction in mice.

Materials and methods

Reproductive toxicity of 100, 200 and 400 mg/kg RS extract were studied in pregnant ICR mice and its offspring. The embryos’ fibroblast growth factor-1 (FGF-1), vascular endothelial growth factor (VEGF) and estrogen receptor-α (ER-α) were evaluated as targets of endocrine/angiogenesis by immunohistochemical test.

Results

The offspring of treated mice (100, 200 and 400 mg/kg RS extract) during their pregnancy had various pathological conditions, suggesting an abnormal FGF signaling phenomenon during pregnancy. Embryos from the 400 mg/kg group had significantly depressed levels of FGF-1 (P < 0.01) and VEGF (P < 0.05) expression levels as compared to controls by immunohistochemical test. Dysplasia in the heart (12.9%), craniofacial region (18.3%) and vertebrae (32.5%) presented in embryos of the 400 mg/kg group. Furthermore, the ER-α expression was inversely proportional to FGF-1 levels in the same embryo (P < 0.01).

Conclusions

These results implicate a FGF signaling abnormality in vivo and indicate that RS has anti-angiogenesis and anti-estrogen toxicity effects in pregnant rodents.     

Aqueous extract of Astragali Radix ameliorates proteinuria in adriamycin nephropathy rats through inhibition of oxidative stress and endothelial nitric oxide synthase

Aim of the study

To investigate the effects of aqueous extract of Astragali Radix (ARE) on the oxidative stress status and endothelial nitric oxide synthase level in adriamycin (ADR) nephropathy rats.

Materials and methods

ADR nephropathy rats were randomly treated with ARE (2.5 g/kg/d, n = 6, ARE group), or benazepril (10 mg/kg/d, n = 6, angiotensin-converting enzyme inhibitor (ACEI) group) for ten weeks. Serum urea nitrogen, creatinine, albumin, total protein, cholesterol and 24-h urinary protein concentration were determined. Renal cortex catalase (CAT), glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), malondialdehyde (MDA) activities, and 24-h urinary NO₃⁻/NO₂⁻ excretion were determined by chromatometry. Renal cortex cyclic guanosine monophosphate (cGMP) level was measured by enzyme immunoassay and eNOS expression was determined by immunohistochemistry.

Results

ARE and ACEI treatments could remarkably reduce more 24 h urinary protein excretion than that in ADR group (88.32 ± 9.96 mg, 81.78 ± 16.28 mg vs. 153.91 ± 28.63 mg, P < 0.01), and there was no difference between ARE and ACEI group. Renal cortex CAT, GSH-Px activities in ARE and ACEI group were significantly higher than ADR group, and renal cortex SOD activity in ARE group was higher than ADR group. Renal cortex MDA activity, cGMP level, and glomerular and tubular eNOS expression in ARE and ACEI group were lower than that in ADR group, and 24-h urinary NO₃⁻/NO₂⁻ excretion in ARE group was lower than ADR group. Renal cortex MDA content (r = 0.895, P < 0.01), cGMP content (r = 0.666, P < 0.01) and eNOS expression in glomerulus (r = 0.910, P < 0.01) were strongly positively associated with 24 h urinary protein excretion. And renal cortex SOD content was negatively associated with 24 h urinary protein excretion (r = −0.861, P < 0.01).

Conclusions

ARE may ameliorate the proteinuria by suppressing the over expression of eNOS, and inhibiting the oxidative injury in ADR nephropathy rats.     

In vitro studies of Gynura divaricata (L.) DC extracts as inhibitors of key enzymes relevant for type 2 diabetes and hypertension

Aim of the study

To evaluate traditionally used herb, Gynura divaricata (L.) DC (Bai Bei San Qi) as in vitro inhibitors of key enzymes involved in the pathogenesis of hyperglycemia and hypertension. We also determined the distribution of enzyme inhibitory activities in different aqueous and non-aqueous extracts.

Materials and methods

The water extract (extract 1) from the aerial parts of Gynura divaricata (L.) was prepared first and then partitioned sequentially with n-butanol, ethyl acetate, and macroporous adsorptive resin (HPD-40) to yield extracts 2-4; the remaining water phase was named extract 5. Angiotensin-1 converting enzyme (ACE), α-amylase α-glycosidase inhibitory activities of the extracts were determined in vitro and chemical composition including total sugar, protein, flavonoid and total alkaloids in the extract were also evaluated.

Results

The water extract of this herb significantly inhibited (p < 0.05) ACE activity (IC₅₀ = 0.37 mg/ml) and showed a moderate potential hypoglycemic effect via in vitro α-amylase (IC₅₀ = 1.36 mg/ml) and α-glycosidase (IC₅₀ = 2.17 mg/ml) inhibition in dose-dependent manner. Further partitioning of the water extract (extracts 2-4) resulted in higher α-amylase inhibitory activities in extract 2 and 3. For α-glycosidase inhibition, extract 3 gave the highest inhibition. ACE inhibitory activities of the extracts were not improved by partitioning. Sugar, protein, flavonoid and alkaloid were found in water extract but only a small portion was partitioned in the n-butanol extract. However, a large portion of the flavonoids and alkaloids were found in ethyl acetate extract.

Conclusion

The results confirmed potential empirical use of Gynura divaricata (L.) DC for the management of hyperglycemia as well as hypertension. The active compounds for inhibition of α-amylase and α-glycosidase inhibition were flavonoids and alkaloids while ACE inhibition probably resulted from synergic effects of all the herb compounds.     

Anti-diabetic activity and potential mechanism of total flavonoids of Selaginella tamariscina (Beauv.) Spring in rats induced by high fat diet and low dose STZ

Aim of the study

To evaluate the anti-diabetic effects of the total flavonoids of Selaginella tamariscina (Beauv.) Spring (TFST), and to explore the pertinent mechanism.

Materials and methods

High fat diet and STZ (35 mg/kg) induced diabetic rats were administered with TFST at graded oral doses (100, 200 and 400 mg/kg/day, ig.) for 8 weeks. A range of parameters, including blood glucose and lipid, serum insulin and glucagon, glucose tolerance, were tested to evaluate its anti-diabetic effects. The determination of protein expression of peroxisome proliferator activated receptor γ (PPAR-γ) in adipose tissue and insulin receptor substrate 1 (IRS-1) in hepatic and skeletal muscle tissues was used to study the mechanism of TFST. Moreover, the preliminary study of TFST on the antioxidant activity was performed.

Results

The TFST possessed anti-diabetic activities as shown by the decreased serum levels of fast blood glucose (FBG), glycosylated hemoglobulin A1C (HbA1c), triglyceride (TG), total cholesterol (TC), free fatty acid (FFA), low density lipoprotein-cholesterol (LDL-C) and glucagon, as well as increased serum levels of high density lipoprotein-cholesterol (HDL-C), insulin and C-peptide. TFST also improved the oral glucose tolerance test (OGTT) to a certain degree. Furthermore, TFST increased the protein expression of PPAR-γ in adipose tissue, and increased the protein expressions of IRS-1 in hepatic and skeletal muscle tissues. These benefits were associated with increased superoxide dismutase (SOD) and decreased malondialdehyde (MDA) in serum.

Conclusions

TFST exert beneficial effects on hyperglycosemia and hyperlipoidemia in diabetic rats possibly through regulating the levers of PPAR-γ in adipose tissue and IRS-1 in hepatic and skeletal muscle tissues.     

Double-blinded randomized controlled trial for immunomodulatory effects of Tulsi (Ocimum sanctum Linn.) leaf extract on healthy volunteers

Ethnopharmacological relevance

Tulsi (Ocimum sanctum Linn.) is considered as a sacred herb and traditionally it is believed that consumption of Tulsi leaf on empty stomach increases immunity. Experimental studies have shown that alcoholic extract of Tulsi modulates immunity.

Materials and Methods

The present study was designed to evaluate the immunomodulatory effects of ethanolic extract of Tulsi leaves through a double-blinded randomized controlled cross-over trial on healthy volunteers. Three hundred milligrams capsules of ethanolic extracts of leaves of Tulsi or placebo were administered to 24 healthy volunteers on empty stomach and the results of 22 subjects who completed the study were analyzed. The primary objective was to study the levels of Th1 and Th2 cytokines (interferon-γ and interleukin-4) during both pre and post intervention period in blood culture supernatants following stimulation with lipopolysaccharide and phytohaemagglutinin. Other immunological parameters such as T-helper and T-cytotoxic cells, B-cells and NK-cells also were analyzed using Flowcytometry.

Results

Statistically significant increase in the levels of IFN-γ (p = 0.039), IL-4 (p = 0.001) and percentages of T-helper cells (p = 0.001) and NK-cells (p = 0.017) were observed after 4 weeks in the Tulsi extract intervention group in contrast to the placebo group.

Conclusions

These observations clearly ascertain the immunomodulatory role of Tulsi leaves extract on healthy volunteers.     

From the vasodilator and hypotensive effects of an extract fraction from Agelanthus dodoneifolius (DC) Danser (Loranthaceae) to the active compound dodoneine

Aim of the study

Effects of the different fractions obtained by partition of ethanolic extract (EE) of Agelanthus dodoneifolius through column chromatography were investigated on rat blood pressure and aortic relaxation and compared to those observed in the presence of crude EE.

Materials and methods

The acute hypotensive activity of EE, fractions and dodoneine, administrated intravenously, was evaluated in anaesthetized rats using the invasive method of blood pressure recording. Bioassay-guided fractionation using rat aorta pre-contracted by norepinephrine to monitor the relaxant activity led to the isolation of dodoneine.

Results

In normotensive rats, injection of EE (0.01-10 mg/kg) produced a dose-dependent decrease in both systolic and diastolic blood pressure without any significant change in heart rate. In a similar way, the EE (0.001-3 mg/mL) caused relaxation of rat pre-contracted aorta in a concentration-dependent manner. Fractionation of the EE afforded 14 fractions, F1-F14, that were tested on rat precontracted aortic rings. At the concentration level of 1 mg/mL, a maximum relaxation effect was observed for fractions F2-F5. F4 was the most effective to elicit a concentration-dependent relaxation effect with an ED₅₀ = 160 ± 1.1 μg/mL (n = 5) and to decreased systolic and diastolic control pressure by 56.9% and 81.6% respectively. F4 contains most of the dihydropyranone dodoneine, with 93% of the sample mass. Dodoneine separated from this fraction was also able to decrease both systolic and diastolic arterial pressure by 32.5% and 38.7% at 100 μg/kg, respectively.

Conclusion

For the first time, this study demonstrates the hypotensive property of the dodoneine present in Agelanthus dodoneifolius.     

Effects on neuroendocrinoimmune network of Lizhong Pill in the reserpine induced rats with spleen deficiency in traditional Chinese medicine

Aim of the study

Lizhong Pill, composed of Radix Ginseng (Panax ginseng C.A. Meyer), Rhizoma Zingiberis (Zingiber officinale Roscoe), Rhizoma Atractylodis Macrocephalae (Atractylodes macrocephala Koidz.) and Radix Glycytthizae (Glycyrrhiza uralensis Fisch.), is a classical herbal product for curing spleen deficiency in traditional Chinese medicine (TCM), and reserpine treated rats show similar signs to TCM spleen deficiency pattern. This paper is aimed to explore the regulatory effect on neuroendocrinoimmune network by Lizhong Pill in reserpine induced TCM spleen deficiency rats.

Materials and methods

100 healthy adult male SD rats, with a mean weight of 200 g, were randomly divided into five groups in average: control group, reserpine treated group, atropine treated group, treatment groups with Lizhong Pill at high dose and low dose (equal to the dosage of crude drugs for 4 g/kg/d and 8 g/kg/d). Rats in reserpine treated group were induced by intraperitoneal injection of reserpine at 0.5 mg/kg d for 4 weeks. The levels of IL-1, IL-6 and gastrin were measured with radioimmunoassay, TNF-α and IFN-γ in serum were measured with ELISA, the level of vasoactive intestinal peptide (VIP) and substance P (SP) in small intestine were determined with radioimmunoassay, and the TNF-α and TGF-β positive cells in small intestine were detected by immunohistological staining. Data were analyzed with SAS 9.1 software package.

Results

The rats in reserpine treated group, body weight, concentrations of IFN-γ, IL-1 and TNF-α in serum, expression of TGF-β in small intestine, VIP in small intestine decreased (P < 0.05), and the level of IL-6 in serum, expression of TNF-α, SP in small intestine and gastrin were increased (P < 0.05). Administration of Lizhong Pill at high dose could increase the body weights at day 21, and the weights of rats in Lizhong Pill groups were much higher compared to reserpine treated group. At high dose of Lizhong Pill could increase the level of TNF-α in serum. Lizhong Pill at high dose and low dose could reverse the changes of IL-1, IL-6 and IFN-γ, gastrin, expression of TGF-β and TNF-α, VIP and SP in small intestine.

Conclusions

The rats treated with reserpine, with similar signs to TCM spleen deficiency, show neuroendocrinoimmune disorders, and the restoration of the neuroendocrinoimmune disorders might be the part of mechanism of Lizhong Pill for reinforcing TCM spleen deficiency.