Hassan Sadraei

Background: Dracocephalum kotschyi is a medicinal plant with well-documented anti-spasmodic and anti-inflammatory properties. However, its potential effects on blood pressure have not been previously investigated. Objective: This study aimed to evaluate the pharmacological effects of D. kotschyi essential oils and its hydroalcoholic extract on blood pressure in rats. Methods: Male Wistar ...

Background and aims: Dracocephalum kotschyi is a medicinal plant native to Iran, which is known for its anti-inflammatory and spasmolytic properties. This study was conducted to investigate the sub-chronic adverse effects of D. kotschyi extract on biochemical and hematological biomarkers in rats, with the aim of identifying any potential adverse effects associated with long-term therapy. Methods: A total of 50 male Wistar rats were randomly allocated into 5 separate groups, each consisting of 10 rats. The extract was administered orally for 90 days at doses of 25, 75, and 300 mg/kg in three distinct groups. A control group and a vehicle-treated group were also included in this study. Following the treatment period, blood samples were collected to assess biochemical and hematological parameters. Liver and kidney tissue samples were prepared for histological examination. Mean comparisons between groups were conducted using the one-way analysis of variance (ANOVA), with P values less than 0.05 considered statistically significant. Results: The hydroalcoholic extract of D. kotschyi at a dose of 300 mg/kg significantly increased levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), and creatinine (Cr). Additionally, ALT and alkaline phosphatase (ALP) levels were significantly elevated at the dose of 75 mg/kg. Other parameters, including electrolytes, glucose, lipid profiles, thyroid hormones, complete blood count (CBC), prothrombin time (PT), and international normalized ratio (INR), remained unaffected, and no tissue damage was observed in the histological examinations. Conclusion: Long-term oral administration of D. kotschyi extract at anti-inflammatory doses in rats resulted in no significant changes in the measured plasma biochemical or hematological biomarkers. However, at higher doses, there was an increase in hepatic biomarkers, as well as LDH and Cr levels, indicating potential organ toxicity associated with overdose.

Introduction: Zaringiah (Dracocephalum kotschyi) is an Iranian endemic herbal plant naturally growing in the Isfahan and Khorasan provinces. Hydroalcoholic extract of Zaringiah has anti-inflammatory, antispasmodic, and immunomodulatory properties. So far, the effect of Zaringiah extract on contraction induced by histamine and serotonin (5-HT) has not been reported. The objective of this research was to investigate the antispasmodic effect of hydroalcoholic, aqueous, chloroform, and ethyl acetate extracts of Zaringiah on rabbit ileum smooth muscle contractions induced by histamine and 5-HT. Methods: Khorasani variant of Zaringiah was used in this study. Aqueous extract was prepared by decoction, while hydroalcoholic extract was obtained by the maceration technique. Chloroform and ethyl acetate fractions were obtained using a solvent in solvent extraction technique. Rabbit isolated ileum was set up in an organ bath filled with Tyrode’s solution. The effects of the above extracts were examined on contractions induced by histamine or 5-HT and compared with each other. Results: Hydroalcoholic extract of Zaringiah inhibited the rabbit ileum contractions induced by histamine (IC50 = 76 ± 7.7 μg/mL) and 5-HT (IC50 = 60 ± 6.4 μg/mL), as well as spontaneous contractions (IC50 = 63 ± 15 μg/mL). The aqueous extract, as well as chloroform and ethyl acetate fractions, concentration-dependently inhibited spontaneous, histamine, and 5-HT induced contractions. Conclusion: There was not a significant difference among the inhibitory actions of hydroalcoholic, aqueous, chloroform, and ethyl acetate extracts of Zaringiah on rabbit ileum, indicating the distribution of active constituents in both polar and non-polar mediums.

Introduction: Zaringiah (Dracocephalum kotschyi) is a famous Iranian herbal plant with anti-inflammatory and spasmolytic activities. There is no standard drug dosage form for the D. kotschyi extract on the market. The objective of this project was to design a suitable oral dosage form for the hydroalcoholic extract of D. kotschyi. Methods: Standard granules were prepared using the moist granulation technique. Physical properties of the granules were determined before filling the capsule with fixed doses of the drug (25 mg and 50 mg). Syrup was prepared in sucrose solution at 5 mg/mL concentration. Bioactivity and phytochemical assays were used for dosage form stability and uniformity evaluations before and after 3- and 6- months incubation. Pharmacological bioassay method was designed to determine the bioactivity of the products before and after incubation. Pharmacological effects of the prepared capsule and syrup were determined on rat isolated ileum and intestinal meal transit, respectively. Results: In this study, D. kotschyi extract was effectively formulated as capsule and syrup for oral consumption. Environmental and aging factors had no significant effect on the total flavonoid or phenolic contents or bioactivity of the manufactured products. Furthermore, the ingredients used in the formulation had no effect on the bioactivity of the active substances in the extract. Conclusion: The standard oral dosage forms prepared from D. kotschyi extract can be used for clinical trials. In addition, we introduced a reliable bioassay technique, which might be applied for the evaluation of herbal medicines with antispasmodic activities.

Introduction: Naringenin is a flavonoid constituent of many herbal plants, including citreous fruits. Biological studies have suggested various therapeutic effects for naringenin, including protective effects on gastrointestinal (GI) motility. The present study was performed to investigate the involvement of ATP-sensitive K+ channels on the effect of naringenin in rat ileum motility. Methods: Ileum contractions were induced by either KCl or acetylcholine (ACh) in vitro. Inhibitory concentration-response curves were constructed for naringenin and diazoxide after exposure of rat isolated ileum to KCl (20mM) or ACh (500nM). The relaxant effects of naringenin and diazoxide were also examined in the presence of glibenclamide. Furthermore, oral effects of diazoxide (25 mg/kg) and naringenin (25, 50 mg/kg) were also assessed on the intestinal charcoal meal transit in mice (n=10) in the absence and presence of glibenclamide (50 mg/kg). Results: Diazoxide and naringenin in a concentration-dependent manner inhibited ileum contractions induced by low bath concentration of KCl (20mM). However, both drugs had no effect on contractions induced by a high concentration of KCl (160mM). The inhibitory effects of diazoxide and naringenin were blocked by glibenclamide. Oral administration of diazoxide and naringenin significantly reduced the intestinal transit of charcoal meal. The delay in the intestinal transit was blocked by the oral dose of glibenclamide. The effect of naringenin on the rat intestinal strip pre-contracted with the KCl was relatively similar to that of ATP-sensitive K+ channel opener (diazoxide). Conclusion: This research supports that ATP-sensitive K+ channels are involved in the rat small intestinal smooth muscles relaxation induced by naringenin.

Introduction: Honeysuckle (Lonicera japonica) is an ornamental flowering plant with numerous traditional medicinal uses. It has been claimed to have anti-spasmodic activities, but only limited studies have been done to support this. The objective of this research was to investigate anti-spasmodic effect of L. japonica flower extract on uterus contractions. Methods: Hydroalcoholic extract was prepared using the maceration technique. In addition, chloroform and ethyl acetate fractions were prepared using a solvent in solvent fractionation technique. Essential oils were collected using the hydro-distillation technique. Rat isolated uterus was suspended in an organ bath and contracted with oxytocin, acetylcholine (ACh), KCl, or application of electrical field stimulation (EFS). The relaxant effects of the extract, its fractions, and nifedipine were examined on uterine contrition induced by the above-mentioned stimuli. Results: Nifedipine in a concentration-dependent manner inhibited uterine contraction induced by oxytocin, KCl, ACh, and EFS. L. japonica flower extract also exhibited an inhibitory effect on the isolated rat uterus. Comparison of the hydroalcoholic extract with its chloroform and ethyl acetate fractions showed that the chloroform fraction was the most potent and the ethyl acetate the weakest part of the plant with antispasmodic activity. The relaxant effect of essential oil had close similarities to that of chloroform extract. Conclusion: Lipophilic compounds isolated by the chloroform partition of crude hydroalcoholic extract of L. japonica flower exhibited the most antispasmodic activity. Ethyl acetate partition of the same extract exhibited the least activity. Therefore, it can be concluded that the spasmolytic constituents of L. japonica flower reside in chloroform partitioning. The nonpolar essential oils may also have a contribution.

Introduction: Dracocephalum kotschyi is an Iranian traditional medicine with anti-inflammatory and bronchodilatory properties. The objective of this research was to investigate the preventive effect of D. kotschyi extract in bleomycin induced fibrosis. Methods: Fibrosis was induced by intratracheal administration of a single dose of bleomycin (5 mg/kg) in rat. One group received daily administration of normal saline. Other groups were treated daily with oral administration of either hydroalcoholic or aqueous extract of D. kotschyi (20, 40, & 80 mg/kg). Another group was treated with pirfenidone (100 mg/kg). After 4 weeks daily treatments, the animals were sacrificed and the whole lungs were dissected out for biochemical and histopathological examinations. The biochemical examination included assessment of hydroxyproline and malondialdehyde levels. Results: The lung tissues in bleomycin treated groups showed severe tissue injuries. However, lung tissues in the groups that received hydroalcoholic or aqueous extracts of D. kotschyi showed mild to moderate tissue injuries. Intratracheal instillation of bleomycin significantly increased hydroxyproline and malondialdehyde biomarker levels compared with the sham group. In the positive control group treated with pirfenidone, there was a marked reduction in both hydroxyproline and malondialdehyde levels. Both hydroalcoholic and aqueous extracts of D. kotschyi significantly prevented the elevation of hydroxyproline and malondialdehyde levels at the above oral doses in comparison to the vehicle treated control group. Conclusion: Prevention of hydroxyproline and malondialdehyde elevation levels in this experiment indicates that D. kotschyi extract might be a suitable remedy for the treatment of respiratory fibrosis induced by drugs such as bleomycin.

Introduction: Quercetin is a flavonoid compound found in many medicinal plants. Antispasmodic effect of quercetin has been reported in ileum and uterus smooth muscles but not in bladder. Therefore, the objective of this research was to investigate relaxant effect of quercetin in rabbit isolated bladder. Methods: Male rabbit was asphyxiated with carbon dioxide and then sacrificed. The whole bladder was dissected out and placed in oxygenated Tyrode’s solution. Isolated bladder was cut into longitudinal strips and placed in an organ bath for contraction studies. Contractions were induced with KCl (20mM), acetylcholine (5μM) and electrical field stimulation (EFS). Full inhibitory concentration–response curve was constructed for quercetin following addition of above spasmogens. Quercetin was added into the organ bath with 2 fold increments in concentration until maximum response was achieved. Nifedipine was used as positive control group and equivalent volume of quercetin vehicle (water + DMSO) was used as negative control group.Results: Quercetin (4 μg/mL to 640 μg/mL) in a concentration dependent manner inhibited isolated bladder strips contracted by KCl (IC50=159±25 μg/mL), acetylcholine (IC50=43±9.1 μg/mL) and EFS (IC50=38±9.3 μg/mL). In the highest used concentration, quercetin completely removed contractile responses to KCl, acetylcholine and electrical filed stimulation (EFS). Nifedipine totally inhibited KCl response (IC50=115±36 ng/mL) but only partially inhibited acetylcholine and EFS responses. Conclusion: These results confirm the relaxant effect of quercetin on rabbit bladder and if similar effects are seen in human studies, then quercetin would be a suitable drug candidate to be investigated for bladder incontinence.

Introduction:Dracocephalum kotschyiextract has antispasmodic activities on smooth muscle including ileum, uterus and trachea. The objective of this research was to investigate antispasmodic activity of hydroalcoholic and flavonoids extracts ofD. kotschyion rabbit bladder contractions.Methods:Rabbits were euthanized by carbon dioxide asphyxiation and the whole bladder was dissected out and immersed in the Tyrode’s solution. Longitudinal bladder strips were mounted vertically in an organ bath at 37°C and gassed continuously with O₂. Bladder strips were contracted with acetylcholine (ACh), KCl, or electrical field stimulation (EFS). Isotonic tension of the tissue was recorded before and after addition of hydroalcoholic or flavonoids rich extracts ofD. kotschyi. Nifedipine and propantheline were used as standard drugs.Results:Standard drug propantheline, prevented bladder phasic contraction induced by ACh (1µM) without affecting KCl response. On the other hand, cumulative addition of nifedipine attenuated the tonic contractions induced by KCl (20mM) on bladder smooth muscle. Hydroalcoholic and flavonoids extracts ofD. kotschyiat concentration ranges of 10-320 µg/ mL in a concentration dependent way inhibited bladder tonic contraction induced by KCl (n=6). Both extracts also in a concentration-dependent manner relaxed EFS and ACh-induced contractions (range, 20–1280 µg/mL) of bladder smooth musclein vitro. Complete inhibition was achieved with the highest used concentrations of the extracts. The inhibitory effect of the extract was reversible following washing the tissues with fresh Tyrode’s solution.Conclusion:This study clearly demonstrated thatD. kotschyiextracts were able to prevent contractions induced by ACh, KCl or EFS in isolated rabbit bladder. This means that people consuming this medicinal plant may face urinary retention which could be a problem for patients with prostate hypertrophy. On the other hand, this plant might be useful in patients with urinary incontinence. However, its usefulness must be assessed in the controlled clinical trials.

Introduction: Pharmacological studies confirm antispasmodic activities of chamomile (Matricaria chamomilla) extract on intestinal smooth muscles and it has been suggested that chamomile increases uterus tone, but so far there is no scientific studies which support this assumption. Therefore, this study was designed to determine spasmodic and spasmolytic activities of M. chamomilla extracts on rat isolated uterus. Methods: Hydro-alcoholic extract of M. chamomilla was prepared by maceration technique. Flavonoids rich extract was prepared by liquid in liquid extraction technique. The spasmodic effects of the extracts were assessed on spontaneously contracting rat uterus. The myorelaxant effect of M. chamomilla extracts was validated on isolated uterus contractions induced by KCl, acetylcholine (ACh), electrical field stimulation (EFS) and oxytocin. Results: Hydro-alcoholic extract of M. chamomilla (0.8 and 1.6 mg/mL) enhanced spontaneous movement of rat isolated uterus smooth muscle suspended in an organ bath. On the other hand, flavonoids rich fraction only diminished uterus contractile activities. Flavonoids rich extract of the plant at bath concentration ranges of 40 μg/mL to 400 μg/mL attenuated uterus response to ACh, KCl, EFS and oxytocin. The hydro-alcoholic extract of M. chamomilla at higher concentration ranges (250 μg/mL to 1.5 mg/mL) inhibited uterus contractions induced by the above spasmogens. Conclusion: The present study confirms both spasmodic and spasmolytic activities M. chamomilla hydro-alcoholic extract. Therefore, medicinal use of the crude extract of M. chamomilla may initiate uterus contraction which could increase risk of spontaneous miscarriage or premature parturition.