Mohd Roslan Sulaiman

Antinociceptive activity of the essential oil of Zingiber zerumbet.

In the present study, the rhizome essential oil from Zingiber zerumbet (Zingiberaceae) was evaluated for antinociceptive activity using chemical and thermal models of nociception, namely, the acetic acid-induced abdominal writhing test, the hot-plate test and the formalin-induced paw licking test. It was demonstrated that intraperitoneal administration of the essential oil of Z. zerumbet (EOZZ) at the doses of 30, 100 and 300 mg/kg produced significant dose-dependent inhibition of acetic acid-induced abdominal writhing, comparable to that of obtained with acetylsalicylic acid (100 mg/kg). At the same doses, the EOZZ produced significant dose-dependent increases in the latency time in the hot-plate test with respect to controls, and in the formalin-induced paw licking test, the EOZZ also significantly reduced the painful stimulus in both neurogenic and inflammatory phase of the test. In addition, the antinociceptive effect of the EOZZ in the formalin-induced paw licking test as well as hot-plate test was reversed by the nonselective opioid receptor antagonist, naloxone suggesting that the opioid system was involved in its analgesic mechanism of action. On the basis of these data, we concluded that the EOZZ possessed both central and peripheral antinociceptive activities which justifying its popular folkloric use to relieve some pain conditions.

A synthetic curcuminoid derivative inhibits nitric oxide and proinflammatory cytokine synthesis

Curcumin is a highly pleiotropic molecule with significant regulatory effects upon inflammation and inflammatory related diseases. However curcumin has one major important limitation in which it has poor bioavailability. Design of synthetic structural derivatives of curcumin is but one approach that has been used to overcome its poor bioavailability while retaining, or further enhancing, its drug-like effects. We have synthesized a series of curcumin analogues and describe the effects of 2,6-bis-4-(hydroxyl-3-methoxy-benzylidine)-cyclohexanone or BHMC upon nitric oxide and cytokine synthesis in cellular models of inflammation. BHMC showed a significant dose-response inhibitory action upon the synthesis of NO and we have shown that this effect was due to suppression of both iNOS gene and enzyme expression without any effects upon scavenging of nitrite. We also demonstrated that BHMC has a very minimal effect upon iNOS activity with no effect at all upon the secretion of PGE(2) but has a strong inhibitory effect upon MCP-1 and IL-10 secretion and gene expression. Secretion and gene expression of TNF-alpha and IL-6 were moderately inhibited whereas IL-8 and IL-1beta were not altered. We conclude that BHMC selectively inhibits the synthesis of several inflammatory mediators. BHMC should be considered a promising drug lead for preclinical and further pharmacological studies.

Zerumbone-Induced Antinociception: Involvement of the l-Arginine-Nitric Oxide-cGMP -PKC-K+ATP Channel Pathways

This study investigated the antinociceptive effects of zerumbone in chemical behavioural models of nociception in mice. Zerumbone given through intraperitoneal route (i.p.) produced dose-related antinociception when assessed on acetic acid-induced abdominal writhing test in mice. In addition, the i.p. administration of zerumbone exhibited significant inhibition of the neurogenic pain induced by intraplantar (i.pl.) injection of capsaicin and bradykinin. Likewise, zerumbone given by i.p. route reduced the nociception produced by i.pl. injection of glutamate and phorbol myristate acetate (PMA). The antinociception caused by zerumbone in the acetic acid test was significantly attenuated by i.p. pre-treatment of mice with l-arginine (nitric oxide precursor) and glibenclamide (ATP-sensitive K(+) channel inhibitor). However, the antinociception of zerumbone was enhanced by methylene blue (non-specific gyanylyl cyclase inhibitor). Together, these results indicate that zerumbone produces pronounced antinociception against chemical models of nociception in mice. It also strongly suggests that the l-arginine-nitric oxide-cGMP-PKC-K(+) ATP channel pathways, the TRPV1 and kinin B2 receptors play an important role in the zerumbone-induced antinociception.

In vitro antiproliferative and antioxidant activities of the extracts of Muntingia calabura leaves.

The in vitro antiproliferative and antioxidant activities of the aqueous, chloroform and methanol extracts of Muntingia calabura leaves were determined in the present study. Assessed using the 3,(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) (MTT) assay, the aqueous and methanol extracts of M. calabura inhibited the proliferation of MCF-7, HeLa, HT-29, HL-60 and K-562 cancer cells while the chloroform extract only inhibited the proliferation of MCF-7, HeLa, HL-60 and K-562 cancer cells. Interestingly, all extracts of M. calabura, which failed to inhibit the MDA-MB-231 cells proliferation, did not inhibit the proliferation of 3T3 (normal) cells, indicating its safety. All extracts (20, 100 and 500 μg/ml) were found to possess antioxidant activity when tested using the DPPH radical scavenging and superoxide scavenging assays with the methanol, followed by the aqueous and chloroform, extract exhibiting the highest antioxidant activity in both assays. The total phenolic content for the aqueous, methanol and chloroform extracts were 2970.4 ± 6.6, 1279.9 ± 6.1 and 2978.1 ± 4.3 mg/100 g gallic acid, respectively. In conclusion, the M. calabura leaves possess potential antiproliferative and antioxidant activities that could be attributed to its high content of phenolic compounds, and thus, needs to be further explored.

Diuretic properties of Orthosiphon stamineus Benth.

ETHNOPHARMACOLOGICAL RELEVANCE Orthosiphon stamineus has been used in traditional medicine for centuries especially to treat diseases of the urinary system. AIM OF THE STUDY To investigate the diuretic activity, to elucidate its possible mechanism and to evaluate the renal effects of Orthosiphon stamineus extract. MATERIALS AND METHODS Water extracts were administered orally at doses of 5 and 10 mg/kg to Sprague-Dawley rats and the control groups were given commercial diuretic drugs either furosemide or hydrochlorthiazide at 10 mg/kg. Urine volume, urine pH, urine density and urine electrolytes were determined every hour for 4h. Blood was assayed for glucose, albumin, blood urea nitrogen (BUN) and creatinine. RESULTS O. stamineus extract exhibited dose-dependent diuretic activity. However, excretion of Na+ and Cl(-) was not markedly elevated, but urinary excretion of K+ was significantly increased. O. stamineus extracts slightly increased the serum BUN, creatinine and blood glucose level. Although these levels were statistically significant when compared to control, these levels were still within normal range. CONCLUSIONS O. stamineus exhibited diuretic activity, but was less potent than furosemide and hydrochlorothiazide. Care should be taken when consuming this herb as slight increase of kidney function enzymes was recorded.

Flavokawain A Induces Apoptosis in MCF-7 and MDA-MB231 and Inhibits the Metastatic Process In Vitro

Introduction The kava-kava plant (Piper methsyticum) is traditionally known as the pacific elixir by the pacific islanders for its role in a wide range of biological activities. The extract of the roots of this plant contains a variety of interesting molecules including Flavokawain A and this molecule is known to have anti-cancer properties. Breast cancer is still one of the leading diagnosed cancers in women today. The metastatic process is also very pertinent in the progression of tumorigenesis. Methods MCF-7 and MDA-MB231 cells were treated with several concentrations of FKA. The apoptotic analysis was done through the MTT assay, BrdU assay, Annexin V analysis, cell cycle analysis, JC-1 mitochondrial dye, AO/PI dual staining, caspase 8/9 fluorometric assay, quantitative real time PCR and western blot. For the metastatic assays, the in vitro scratch assay, trans-well migration/invasion assay, HUVEC tube formation assay, ex vivo rat aortic ring assay, quantitative real time PCR and western blot were employed. Results We have investigated the effects of FKA on the apoptotic and metastatic process in two breast cancer cell lines. FKA induces apoptosis in both MCF-7 and MDA-MB231 in a dose dependent manner through the intrinsic mitochondrial pathway. Additionally, FKA selectively induces a G2/M arrest in the cell cycle machinery of MDA-MB231 and G1 arrest in MCF-7. This suggests that FKAs anti-cancer activity is dependent on the p53 status. Moreover, FKA also halted the migration and invasion process in MDA-MB231. The similar effects can be seen in the inhibition of the angiogenesis process as well. Conclusions FKA managed to induce apoptosis and inhibit the metastatic process in two breast cancer cell lines, in vitro. Overall, FKA may serve as a promising candidate in the search of a new anti-cancer drug especially in halting the metastatic process but further in vivo evidence is needed.

Antinociceptive, anti-inflammatory and antipyretic effects of Muntingia calabura aqueous extract in animal models

The present study was carried out to elucidate the potential of aqueous extract of Muntingia calabura leaves aqueous extract (MCAE) as antinociceptive, anti-inflammatory and antipyretic agents using the formalin-, carrageenan-induced paw edema- and brewer’s yeast-induced pyrexia tests in rats. The extract was prepared by soaking the dried powdered leaves of M. calabura in distilled water (dH2O) overnight. The supernatant obtained, considered as a stock solution (100% concentration/strength), was then diluted to concentrations of 10% and 50% and used together in all experimental models. The MCAE, at concentrations of 10%, 50% and 100%, were found to show significant antinociceptive, anti-inflammatory and antipyretic activities in all tests. However, all of the activities occurred in a concentration-independent manner. The 50% and 100% concentrations of MCAE produced insignificant antinociceptive and antipyretic activities, respectively. Although the 100% concentration of MCAE produced significant (P<0.05) anti-inflammatory activity, the activity was lower than that of the 10% and 50% concentrations of MCAE. Based on the results, we conclude that the M. calabura leaves possessed antinociceptive, anti-inflammatory and antipyretic activities and, thus, justifies the Peruvian folklore claims of its medicinal values.

The antinociceptive action of aqueous extract from Muntingia calabura leaves: the role of opioid receptors.

Objective: The present study was carried out to investigate the antinociceptive activity of the aqueous extract of Muntingia calabura (MCAE) leaves and to determine the effect of temperature and the involvement of the opioid receptor on the said activity using the abdominal constriction test (ACT) and hot-plate test (HPT) in mice. Materials and Methods: The extract was prepared by soaking the dried powdered leaves of M. calabura in distilled water (dH2O) overnight, and the supernatant obtained was considered as a stock solution with 100% concentration. The stock solution was diluted to 1, 5, 10, 50 and 100% and used to determine the antinociceptive activity of MCAE. A further experiment was done with 50% concentration to determine the effect of temperature and naloxone involvement of the opioid receptor system in MCAE antinociceptive activity. Results: At the various concentrations MCAE showed significant antinociceptive activity in both tests. However, the concentration-dependent activity was observed only in the ACT but not in the HPT. The 50% concentration of MCAEs were also stable against the effect of various temperatures as indicated by the presence of activity in both tests. The temperatures (40, 60 and 100°C) also showed an enhanced extract activity only in the HPT. Pre-treatment with naloxone (2 and 10 mg/kg) blocked the extract activity in both tests, indicating the involvement of the opioid receptor system in MCAE antinociceptive activity. Conclusion: Our data indicate that M. calabura leaves possess antinociceptive activity against chemically and thermally induced noxious stimuli. The bioactive compound(s) responsible for its antinociceptive activity is/are heat-stable and work partly via the opioid receptor system.

Antinociceptive, Anti-Inflammatory and Antipyretic Properties of the Aqueous Extract of Bauhinia purpurea Leaves in Experimental Animals

Objective: The present study was carried out to determine the antinociceptive, anti-inflammatory and antipyretic activities of the aqueous extract of Bauhinia purpurea leaves using animal models. Materials and Methods: The air-dried, powdered leaves (≈20 g) were soaked in distilled water (1:20; w/v) at room temperature for 72 h and the supernatant obtained was freeze-dried. The crude dried extract (≈2.4 g) was prepared in doses of 6.0, 30.0 and 60.0 mg/kg, and subjected to the respective antinociceptive (abdominal constriction, hot plate and formalin tests), anti-inflammatory (carrageenan-induced paw edema test) and antipyretic (brewer’s yeast-induced pyrexia test) assays. Results:The results obtained indicate that the extract possessed significant (p < 0.05) antinociceptive, anti-inflammatory and antipyretic activities, which were not dependent on the doses of extract used. The highest concentration of extract was less effective as an anti-inflammatory and an antipyretic agent. Conclusion: This study showed that the aqueous extract of B. purpurea leaves possesses potential pharmacological activities that require further investigation and, thus, confirms the folklore use of the plant in the treatment of ailments associated with pain and inflammation.

Possible participation of nitric oxide/cyclic guanosine monophosphate/protein kinase C/ATP-sensitive K(+) channels pathway in the systemic antinociception of flavokawin B.

The possible mechanisms of action in the antinociceptive activity induced by systemic administration (intraperitoneal, i.p.) of flavokawin B (FKB) were analysed using chemical models of nociception in mice. It was demonstrated that i.p. administration of FKB to the mice at 0.3, 1.0, 3.0 and 10 mg/kg produced significant dose-related reduction in the number of abdominal constrictions. The antinociception induced by FKB in the acetic acid test was significantly attenuated by i.p. pre-treatment of mice with L-arginine, the substrate for nitric oxide synthase or glibenclamide, the ATP-sensitive K(+) channel inhibitor, but was enhanced by methylene blue, the non-specific guanylyl cyclase inhibitor. FKB also produced dose-dependent inhibition of licking response caused by intraplantar injection of phorbol 12-myristate 13-acetate, a protein kinase C activator (PKC). Together, these data indicate that the NO/cyclic guanosine monophosphate/PKC/ATP-sensitive K(+) channel pathway possibly participated in the antinociceptive action induced by FKB.