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

作　　者：Paween Kunsorn Nijsiri Ruangrungsi Vimolmas Lipipun Ariya Khanboon Kanchana Rungsihirunrat 

机构地区：[1]College of Public Health Sciences,Chulalongkorn University,Bangkok 10330,Thailand [2]Faculty of Pharmaceutical Sciences,Chulalongkorn University,Bangkok 10330,Thailand

出　　处：《Asian Pacific Journal of Tropical Biomedicine》2013年第4期284-290,共7页亚太热带生物医学杂志（英文版）

基　　金：Supported by Office of the Higher Education Commission;Thailand;University of Phayao and CU Graduate School Thesis Grant;Chulalongkorn University (Grant No.5200601)

摘　　要：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 PGR 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.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±0.81and 3.37±0.31,respectively.The PCR amplification of ITS region generated the PGR product approximately 700 bp in size.There were 34 polymorphisms within the ITS region which consisted of 11 Indels and 23 nucleotide substitutions.The IC_(50)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.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.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 PGR 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.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±0.81and 3.37±0.31,respectively.The PCR amplification of ITS region generated the PGR product approximately 700 bp in size.There were 34 polymorphisms within the ITS region which consisted of 11 Indels and 23 nucleotide substitutions.The IC50values 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.Conclusions:The combination of macroscopic,microscopic and biomolecular method are able to authenticate these closely related plants and both of

关 键 词：Clinacanthus nutans Clinacanthus siamensis Microscopic ANALYSIS Biomolecular ANALYSIS Internal transcribed SPACER HERPES SIMPLEX virus PLAQUE reduction assay 

分 类 号：R373[医药卫生—病原生物学]