The composition of the ethanolic extract from the aerial parts of Crocus alatavicus Regel & Semen from southern Kazakhstan spontaneous flora was analyzed together with the determination of its antibacterial, antifungal, antiviral and anticancer activity. The phytochemical profile analysis by high-performance liquid chromatography-electrospray ionization-quadrupole-time of flight-mass spectrometry (HPLC/ESI-QTOF-MS) revealed the presence of multiple kaempferol derivatives. High-performance reverse-phase liquid chromatography combined with a photodiode-array detection (RP-HPLC/PDA) found that kaempferol 3-O-dihexoside and kaempferol 3-O-acyltetrahexoside accounted for 70.5% of the kaempferol derivatives. The minimum inhibitory concentration (MIC) values of the extract for all the tested reference microorganisms were high, reaching 10 mg/mL for yeasts and 20 mg/mL for bacteria. In contrast, antiviral activity was observed at 2 mg/mL, resulting in the inhibition of the HSV-1-induced cytopathic effect and the reduction in virus infectious titer by 1.96 log, as well as the viral load by 0.85 log. Among the tested prostate cancer cell lines, significant cytotoxic activity of the extract was noted only on the LNCaP cell line, with an IC50 value of 1.95 mg/mL. The LNCaP cell line treated with 2 mg/mL of the extract showed a noticeably reduced number of spindle-shaped cells with longer cellular projections, a significant increase in the peak corresponding to the population of apoptotic cells in the sub-G1 phase and a decreased intracellular glutathione (GSH) level, suggesting the prooxidative properties of the extract. The obtained data provide novel information about the flavonoids present in the aerial part of C. alatavicus and suggest its potential application as a source of the compounds active against HSV-1 and metastatic, androgen-sensitive prostate cancer.
Plants of the genus Crocus L. have a wide spectrum of pharmacological action. To date, many of their species have not been fully studied. Morphological and anatomical features of Crocus alatavicus Regel & Semenow have not been studied and there is no regulatory documentation. The literature review indicates the absence of a specific, complete description of these features that make it possible to identify and standardize Crocus alatavicus raw materials. The aim of this study is to research the features of morphological and anatomical diagnostic features of the plant C. alatavicus. Morphology of raw materials were studied using a binocular magnifier. Anatomical and diagnostic signs were determined by preparing temporary preparations according to the pharmacopoeia technique using an MS-300 microscope with a camera. Morphological and anatomical features of the reduced stem, leaves, and flower were studied. It has been established that the leaves of C. alatavicus have a unique and peculiar shape in cross-section, like most species of Crocus L. It consists of a central quadrangular "keel" and two side "arms". C. alatavicus differs from other species in the triangular shape of the "keel" and the tips of the "arms" are strongly curved to the "keel". There is a pale stripe in the axial direction along the center of the "keel". The leaves of C. alatavicus are amphistomatic in the arrangement of stomata and belong to the anomocytic type. These signs of plant are used to identify medicinal plant raw materials. It is important to emphasize that the standardization of C. alatavicus domestic raw materials creates new opportunities for its further use in medicine. Сrосиѕ L. тұқымдасының өсімдіктері фармакологиялық әсердің кең спектріне ие, олардың көптеген түрлері бүгінгі күнге дейін аз зерттелген. Олардың ішінде шикізатқа нормативтік құжаттама жетіспейтін Crocus alatavicus Regel & Semenow бар, өйткені бұл өсімдіктің морфологиялық-анатомиялық белгілері зерттелмеген. Әдебиеттерге шолу Crocus alatavicus шикізатын анықтауға және стандарттауға мүмкіндік беретін осы белгілердің нақты, толық сипаттамасының жоқтығын көрсетті. Зерттеудің мақсаты C. alatavicus шикізатының морфологиялық және анатомиялық-диагностикалық белгілерін зерттеу. Шикізаттың морфологиялық белгілері бинокулярлық лупаның көмегімен зерттелді, анатомиялық-диагностикалық белгілер фармакопеялық әдіске сәйкес уақытша препараттарды дайындау арқылы фотокамералы МС-300 микроскопын қолдана отырып анықталды. Қысқартылған сабақтың, жапырақтың, гүлдің морфологиялық және анатомиялық-диагностикалық белгілері зерттелді. C. alatavicus жапырағының көлденең қимасы Crocus L. тусының басқа түрлері сияқты ерекше пішінге ие екені анықталды. Ол Орталық төртбұрышты "кильден"және екі бүйірлік "қолдардан" тұрады. Алатау шафранының зерттелген басқа Crocus L. түрлерінен айырмашылығы, "киль" үшбұрыш пішінді және "қолдарының" ұштары "кильге" өте иілген болып келеді. Осьтік бағытта "киль" орталығының бойында бозғылт жолақ бар. Алатау шафранында лептесіктердің орналасуы бойынша жапырақтары амфистоматикалық, лептесіктері – аномоциттік типті болып келеді. Өсімдік шикізатының осы белгілері шикізатты сәйкестендіру үшін қолданылады және дәрілік өсімдік шикізатын стандарттаудың маңызды аспектілерінің бірі болып табылады. Растения рода Сrocus L. обладают широким спектром фармакологического действия, многие их виды на сегодняшний день мало изучены. В их числе Crocus alatavicus Regel & Semenow, на сырье которого отсутствует нормативная документация, так как не изучены морфолого-анатомические признаки этого растения. Обзор литературы свидетельствует об отсутствии конкретного, полного описания этих признаков, которые делают возможным идентификацию и стандартизацию сырья Crocus alatavicus. Целью настоящего исследования является изучение морфологических и анатомо-диагностических признаков растение C. alatavicus. Морфологические признаки сырья изучали с помощью бинокулярной лупы, анатомо-диагностические признаки определяли путем приготовления временных препаратов согласно фармакопейной методике, используя микроскоп МC-300 c фотокамерой. Изучены морфологические и анатомо-диагностические признаки редуцированного стебля, листьев, цветка. Установлено, что листья C. alatavicus, как и большинство видов Crocus L. в поперечном сечении имеет уникальную и своеобразную форму. Состоит из центрального четырехугольного «киля» и двух боковых «рук». В отличие от других изученных видов Crocus L., у C. alatavicus «киль» имеет трехугольную форму и кончики «рук» очень изогнуты к «килю». В осевом направлении вдоль центра «киля» имеется бледная полоса. По расположению устьиц листья C. alatavicusназываются амфистоматическими, по структуре относиться аномоцитному типу. Данные признаки растительного сырья применяется для идентификации сырья, и являются одним из важных аспектов стандартизации лекарственного растительного сырья.
According to the forecasts of the World Health Organization, the share of herbal medicines in the global pharmaceutical market is more than 40%. The pronounced growth trend of phytopreparations can lead to depletion of stocks of medicinal plants and the introduction of medicinal plants into the culture is a suitable solution to the problem. In order to ensure the quality of medicinal plants, in 2003 the World Health Organization (WHO) issued guidelines on Good cultivation and harvesting practices (GACP) medicinal plants. This article presents the results of the introduction of the Crocus alatavicus plant by seeds. The aim of the study is to develop a technology for the introduction of Crocus alatavicus by the method of seed propagation according to the principles of GACP. The development of the introduction technology according to the principles of GACP guarantees the provision of domestic pharmaceutical production with high-quality plant raw materials. The introduction was carried out at the pilot production plantation of "Phytoleum" LLP. The results of the study showed that during seed propagation, autumn sowing of Crocus alatavicus seeds is preferable. The introduction of organic fertilizer of vermicompost before sowing seeds has a positive effect on the germination of seeds and the survival of seedlings. The plant in the fourth year of vegetation goes into the generative period and the fruit will form. The results of the introduction of Crocus alatavicus by seeds in the period from 2018 to 2022 showed good seed germination and high survival of seedlings, which makes it possible to recommend the plant for introduction into culture. It is important to emphasize that the ability to cultivate C. alatavicus from seeds creates new opportunities for use as domestic raw materials for the purpose of further rational use in scientific and practical medicine, preserving this species in culture.
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