The identities and anti-herpes simplex virus activity of Clinacanthus nutans and Clinacanthus siamensis

<p><b>OBJECTIVE</b>To distinguish the difference among the Clinacanthus nutans (Burm. f.) Lindau (C. nutans) and Clinacanthus siamensis Bremek (C. siamensis) by assessing pharmacognosy characteristics, molecular aspect and also to evaluate their anti-herpes simplex virus (HSV) type 1 and type 2 activities.</p><p><b>METHODS</b>Macroscopic and microscopic evaluation were performed according to WHO Geneva guideline. Stomatal number, stomatal index and palisade ratio of leaves were evaluated. Genomic DNA was extracted by modified CTAB method and ITS region was amplified using PCR and then sequenced. Dry leaves were subsequently extracted with n-hexane, dichloromethane and methanol and antiviral activity was performed using plaque reduction assay and the cytotoxicity of the extracts on Vero cells was determined by MTT assay.</p><p><b>RESULTS</b>Cross section of midrib and stem showed similar major components. Leaf measurement index of stomatal number, stomatal index and palisade ratio of C. nutans were 168.32±29.49, 13.83±0.86 and 6.84±0.66, respectively, while C. siamensis were 161.60±18.04, 11.93±0.81 and 3.37±0.31, respectively. The PCR amplification of ITS region generated the PCR product approximately 700 bp in size. There were 34 polymorphisms within the ITS region which consisted of 11 Indels and 23 nucleotide substitutions. The IC50 values of C. nutans extracted with n-hexane, dichloromethane and methanol against HSV-1 were (32.05±3.63) µg/mL, (44.50±2.66) µg/mL, (64.93±7.00) µg/mL, respectively where as those of C. siamensis were (60.00±11.61) µg/mL, (55.69±4.41) µg/mL, (37.39±5.85) µg/mL, respectively. Anti HSV-2 activity of n-hexane, dichloromethane and methanol C. nutans leaves extracts were (72.62±12.60) µg/mL, (65.19±21.45) µg/mL, (65.13±2.22) µg/mL, respectively where as those of C. siamensis were (46.52±4.08) µg/mL, (49.63±2.59) µg/mL, (72.64±6.52) µg/mL, respectively.</p><p><b>CONCLUSIONS</b>The combination of macroscopic, microscopic and biomolecular method are able to authenticate these closely related plants and both of them have a potency to be an anti-HSV agent.</p>

The identities and anti-herpes simplex virus activity of Clinacanthusnutans and Clinacanthus siamensis

Objective: To distinguish the difference among the Clinacanthus nutans (Burm. f.) Lindau (C. nutans) and Clinacanthus siamensis Bremek (C. siamensis) by assessing pharmacognosy characteristics, molecular aspect and also to evaluate their anti-herpes simplex virus (HSV) type 1 and type 2 activities. Methods: Macroscopic and microscopic evaluation were performed according to WHO Geneva guideline. Stomatal number, stomatal index and palisade ratio of leaves were evaluated. Genomic DNA was extracted by modified CTAB method and ITS region was amplified using PCR and then sequenced. Dry leaves were subsequently extracted withn-hexane, dichloromethane and methanol and antiviral activity was performed using plaque reduction assay and the cytotoxicity of the extracts on Vero cells was determined by MTT assay. Results: Cross section of midrib and stem showed similar major components. Leaf measurement index of stomatal number, stomatal index and palisade ratio of C. nutans were 168.32±29.49, 13.83±0.86 and 6.84±0.66, respectively, while C. siamensis were 161.60±18.04, 11.93±0.81 and 3.37±0.31, respectively. The PCR amplification of ITS region generated the PCR product approximately 700 bp in size. There were 34 polymorphisms within the ITS region which consisted of 11 Indels and 23 nucleotide substitutions. The IC50 values of C. nutans extracted with n-hexane, dichloromethane and methanol against HSV-1 were (32.05±3.63) μg/mL, (44.50±2.66) μg/mL, (64.93±7.00) μg/mL, respectively where as those of C. siamensis were (60.00±11.61) μg/mL, (55.69±4.41) μg/mL, (37.39±5.85) μg/mL, respectively. Anti HSV-2 activity of n-hexane, dichloromethane and methanolC. nutans leaves extracts were (72.62±12.60) μg/mL, (65.19±21.45) μg/mL, (65.13±2.22) μg/mL, respectively where as those of C. siamensis were (46.52±4.08) μg/mL, (49.63±2.59) μg/mL, (72.64±6.52) μg/mL, respectively. Conclusions: The combination of macroscopic, microscopic and biomolecular method are able to authenticate these closely related plants and both of them have a potency to be an anti-HSV agent.

α-Glucosidase Inhibitory Activity of Water Soluble Extract from Thai Mimosaceous Plants

The polar part of twenty species of Thai plants in Mimosaceae family was studied for their α-glucosidase inhibition activity by spectrophotometry. The result showed that most water soluble parts of Albizia lebbeck (L.) Benth branch, Xylia xylocarpa (Roxb.) Taub. bark, Archidendron jiringa I.C. Nielsen seed coat, Albizia lebbeck (L.) Benth. branch bark and Parkia speciosa Hassk pericarb showed high α-glucosidase inhibition with 77.90, 75.62, 74.09, 68.10 and 61.86 percent respectively whilst their half inhibition concentrations (IC50 ) were 0.1554, 0.3143, 0.3829, 0.3965 and 0.4104 mg/ml respectively

Quantitative analysis of oxyresveratrol content in Artocarpus lakoocha and 'Puag-Haad'

To develop a thin-layer chromatography [TLC] densitometric method for the determination of oxyresveratrol content in Artocarpuslakoocha heartwood and in the traditional drug 'Puag-Haad'. Sample solution of A. lakoocha heartwood was prepared by Soxhlet extraction of the plant material in ethanol, whereas the Puag-Haad solution was obtained by dissolving the drug in methanol. Analysis of each sample solution was performed on a Silica gel 60 F254 TLC plate [20 x 10 cm] with methylene chloride/methanol [85:15] as the mobile phase. After development, the TLC plate was examined with a TLC scanner in the absorbance mode at 254 nm. The newly developed analytical method was validated using an authentic sample of oxyresveratrol previously isolated from A. lakoocha heartwood, and was used to analyze the oxyresveratrol content in samples of A. lakoocha heartwood and the traditional drug Puag-Haad. A sensitive and reliable TLC densitometric method was successfully developed. The method was validated in terms of accuracy [99.11-102.60%] and precision [1.66-4.23% coefficient of variation]. The limits of detection and quantitation were 15.6 and 52 ng/spot, respectively. The amounts of oxyresveratrol in 3 samples of A. lakoocha heartwood collected from its natural habitat were 49.0-182.3 mg/g, whereas those in 11 commercial samples were in the range of 23.4-69.6 mg/g. The oxyresveratrol contents in 2 samples of traditional drug Puag-Haad were 780.1 and 837.5 mg/g. The TLC densitometric method developed in this study is a simple, convenient, sensitive and reliable procedure. It was an effective analytical tool for the evaluation of oxyresveratrol content in both A. lakoocha heartwood and the traditional drug Puag-Haad