鳄嘴花和暹罗鳄嘴花的特性及抗单纯疱疹病毒活性
The identities and anti-herpes simplex virus activity of Clinacanthus nutans and Clinacanthus siamensis.
作者信息
Kunsorn Paween, Ruangrungsi Nijsiri, Lipipun Vimolmas, Khanboon Ariya, Rungsihirunrat Kanchana, Chaijaroenkul Wanna
机构信息
College of Public Health Sciences, Chulalongkorn University, Bangkok 10330, Thailand.
出版信息
Asian Pac J Trop Biomed. 2013 Apr;3(4):284-90. doi: 10.1016/S2221-1691(13)60064-7.
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 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±0.81 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.
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.
目的
通过评估生药学特征、分子层面特征来区分鳄嘴花（Clinacanthus nutans (Burm. f.) Lindau，简称C. nutans）和暹罗鳄嘴花（Clinacanthus siamensis Bremek，简称C. siamensis），并评估它们对1型和2型单纯疱疹病毒（HSV）的活性。
方法
根据世界卫生组织日内瓦指南进行宏观和微观评估。评估叶片的气孔数、气孔指数和栅栏组织比。采用改良CTAB法提取基因组DNA，利用聚合酶链反应（PCR）扩增ITS区域，然后进行测序。随后用正己烷、二氯甲烷和甲醇提取干燥叶片，并采用蚀斑减少试验检测抗病毒活性，通过MTT试验测定提取物对非洲绿猴肾细胞（Vero细胞）的细胞毒性。
结果
中脉和茎的横切面显示主要成分相似。鳄嘴花的叶片气孔数、气孔指数和栅栏组织比测量指标分别为168.32±29.49、13.83±0.86和6.84±0.66，而暹罗鳄嘴花分别为161.60±18.04、11.93±0.81和3.37±0.31。ITS区域的PCR扩增产生了大小约为700 bp的PCR产物。ITS区域内有34个多态性位点，包括11个插入缺失和23个核苷酸替换。鳄嘴花用正己烷、二氯甲烷和甲醇提取的提取物对HSV-1的半数抑制浓度（IC50）值分别为(32.05±3.63) μg/mL、(44.50±2.66) μg/mL、(64.93±7.00) μg/mL，而暹罗鳄嘴花的分别为(60.00±11.61) μg/mL、(55.69±4.41) μg/mL、(37.39±5.85) μg/mL。鳄嘴花正己烷、二氯甲烷和甲醇叶提取物对HSV-2的活性分别为(72.62±12.60) μg/mL、(65.19±21.45) μg/mL、(65.13±2.22) μg/mL，而暹罗鳄嘴花的分别为(46.52±4.08) μg/mL、(49.63±2.59) μg/mL、(72.64±6.52) μg/mL。
结论
宏观、微观和生物分子方法相结合能够鉴别这些近缘植物，且二者均有成为抗HSV药物的潜力。
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