丹参、雪山鼠尾、云南鼠尾的显微鉴定学、细胞学及药用成分、无机元素含量研究
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鼠尾草属(Salvia Lim．)植物隶属于唇形科(Lamiaceae)鼠尾草族(Salvieae)，是唇形科中最大的属，全世界约有1050余种。在热带和温带都有分布，主要分布于地中海沿岸、中亚、墨西哥高地以及南美的安第斯山脉等。我国约有78种、24变种和8变型，分布于我国大部分地区，尤以西南为最多，其中有药用价值的植物有33种(含变种、变型)，在民间可作丹参入药的有26种，如云南鼠尾草，雪山鼠尾草等。 丹参(Salvia mihiorrhiza Bunge．)是鼠尾草属多年生直立草本植物，是历版《中华人民共和国药典》收载的中药丹参的唯一来源，又称血参、紫丹参等，作为传统中药在我国沿用已久，始载于《神农本草经》，被列为上品，具有祛瘀止痛、活血通经、清心除烦的功效，主治冠心病、心绞痛、心烦不眠、月经不调等症。野生丹参常生于低山、丘陵地带的路旁、坡地及溪谷旁，海拔120～1300m，在我国主要分布于黄河流域、长江流域各省及西南地区，主要产地有四川、河南、山东、浙江、江苏、湖北、安徽、河北、陕西及湖南等。其中四川中江近百年的栽培丹参，以其色朱、味浓而驰名中外。 本文拟通过中药鉴定学、细胞学和药用成分、无机元素含量的测定等研究方面，探讨鼠尾草属丹参类植物资源性状和显微鉴定特征、染色体数目和倍性及药用成分和无机元素的含量等，旨在为筛选优质的丹参种质资源，为建立评价丹参植物资源优劣的质量指标体系提供理论依据。主要研究结果如下:(1)性状和显微鉴别研究结果显示:来自我国六省的丹参性状和显微特征与《中药鉴定学》中对药材丹参的描述一致。在药材性状上，雪山鼠尾草枝根多，根部呈辫子状扭曲，色朱味浓；云南鼠尾草根上具密集的叶柄痕，支根分枝处变细，呈纺锤形。在显微鉴别上，雪山鼠尾草和云南鼠尾草木质部不成束，导管数目较少，散布，无中央纤维束。为雪山鼠尾草、云南鼠尾草和丹参正品的鉴别提供了依据。研究还发现四川、山东、河南丹参根的皮层比较发达，据此分析认为与丹参酮ⅡA的含量相关；山东、河南、湖北、浙江等地丹参木质部较发达，导管数较多，与水溶性成分含量相关。 (2)细胞学的研究结果显示:不同产地的丹参和云南鼠尾草染色体数目均为2n=2x=16；雪山鼠尾草染色体为2n=4x=32；浙江丹参的体细胞中观察到一条染色体具随体。染色体形态均较小，绝对长度范围0.5～2.0gm，平均长度范围0.7～1.5 gm，属于微小型和小型染色体，很难用于核型分析。山东泰山丹参和四川中江丹参的体细胞内存在天然四倍体细胞，染色体数目为2n=4x=32，初步分析认为与药用成分的含量多少有关。 (3)药用成分含量的HPI。C测定结果显示:河南南阳和四川中江两个道地产区的丹参中丹参酮IIA的含量较高，分别为4268μg/g和4094μg/g；浙江嵊州和四川中江丹参中丹酚酸B的含量较高，分别为34.62 mg/g和28.57 mg/g；湖北大别山和山东泰山丹参中原儿茶醛含量较高分别为939.8μg/g和897.8μg/g。雪山鼠尾草中丹参酮ⅡA的含量符合《中华人民共和国药典》规定指标，可以作为丹参酮ⅡA来源药材。云南鼠尾草中丹参酮ⅡA、丹酚酸B和原儿茶醛的含量均较低，对其药用价值应作进一步探讨。 (4)10种无机元素含量的测定结果显示:浙江嵊州丹参中Ca，Mg，Fe，Mo，Cu，Cd和Pb的含量均为最高分别为11.07 mg/g、10.73 mg/g.333 mg/g1.447μg/g、27.03μg/g、0.2610μg/g和2.165μg/g，对微量元素有较强的富集能力，其中元素Cu的含量超过了《中华人民共和国药典》的指标，有害元素Cd和Pb的含量也较高，不适合作为主流丹参药材在市场上推广；云南鼠尾草中Cu和Pb元素的含量仅次于浙江丹参，分别为0.1850μg/g和1.103μg/g。
The genus Salvia (Lim.) belongs to the tribe Salvieae of the family Lamiaceae, and is the largest genus in Lamiaceae, with approximately 1,050 species worldwide. It is distributed in both tropical and temperate regions, mainly along the Mediterranean coast, Central Asia, the Mexican Highlands, and the Andes Mountains in South America. In China, there are about 78 species, 24 varieties, and 8 forms, distributed across most parts of China, especially concentrated in the southwest. Among them, 33 species (including varieties and forms) have medicinal value, and 26 species can be used as substitutes for Danshen in folk medicine, such as Salvia yunnanensis and Salvia bulleyana. Salvia miltiorrhiza Bunge is a perennial erect herb in the genus Salvia, and is the only source of the traditional Chinese medicine Danshen recorded in all editions of the Pharmacopoeia of the People's Republic of China. It is also known as Xueshen, Zidanshen, etc. It has been used as a traditional Chinese medicine in China for a long time, first recorded in *Shennong Bencao Jing and listed as top-grade. It has the effects of promoting blood circulation to resolve stasis, relieving pain, stimulating menstrual flow, and clearing heart fire to relieve restlessness, and is indicated for coronary heart disease, angina pectoris, insomnia, irregular menstruation, etc. Wild Salvia miltiorrhiza often grows on roadsides, slopes, and along valleys in low mountains and hilly areas, at an altitude of 120–1300 m. In China, it is mainly distributed in provinces along the Yellow River and Yangtze River basins and the southwest region, with main producing areas including Sichuan, Henan, Shandong, Zhejiang, Jiangsu, Hubei, Anhui, Hebei, Shaanxi, and Hunan. The cultivated Danshen in Zhongjiang, Sichuan has been grown for nearly a century, renowned worldwide for its deep red color and strong taste. This paper aims to explore the morphological and microscopic identification characteristics, chromosome number and ploidy, as well as the contents of medicinal components and inorganic elements of Salvia Danshen group plants, through studies in the fields of traditional Chinese medicine identification, cytology, and determination of medicinal components and inorganic elements, with the purpose of screening high-quality Danshen germplasm resources and providing a theoretical basis for establishing a quality index system to evaluate the quality of Danshen plant resources. The main research results are as follows: (1) Results of morphological and microscopic identification: The morphological and microscopic characteristics of Danshen from six provinces in China are consistent with the description of Danshen in *Identification of Traditional Chinese Medicine. In terms of morphological characteristics: Salvia bulleyana has numerous fibrous roots, which are twisted into braid-shaped, deep red in color with strong taste; Salvia yunnanensis has dense petiole scars on the roots, with thin roots at the branching points, spindle-shaped. In microscopic identification: The xylem of Salvia bulleyana and Salvia yunnanensis is not bundled, with fewer scattered vessels, and no central fiber bundle, providing a basis for the identification of Salvia bulleyana, Salvia yunnanensis and genuine Danshen. It was also found that the cortex of Danshen roots from Sichuan, Shandong, and Henan is well-developed, which is preliminarily analyzed to be related to the content of tanshinone ⅡA; the xylem of Danshen from Shandong, Henan, Hubei, and Zhejiang is well-developed with more vessels, which is related to the content of water-soluble components. (2) Cytological results: The chromosome numbers of Danshen from different producing areas and Salvia yunnanensis are all 2n=2x=16; the chromosome number of Salvia bulleyana is 2n=4x=32; one chromosome with a satellite was observed in the somatic cells of Danshen from Zhejiang. All chromosomes are small, with absolute lengths ranging from 0.5 to 2.0 μm, average lengths ranging from 0.7 to 1.5 μm, belonging to micro and small chromosomes, making karyotype analysis difficult. Natural tetraploid cells were observed in the somatic cells of Danshen from Mount Tai, Shandong and Zhongjiang, Sichuan, with chromosome number 2n=4x=32, which is preliminarily analyzed to be related to the content of medicinal components. (3) Results of HPLC determination of medicinal component contents: The contents of tanshinone ⅡA in Danshen from two genuine producing areas of Nanyang, Henan and Zhongjiang, Sichuan are relatively high, 4268 μg/g and 4094 μg/g respectively; the contents of salvianolic acid B in Danshen from Shengzhou, Zhejiang and Zhongjiang, Sichuan are relatively high, 34.62 mg/g and 28.57 mg/g respectively; the contents of protocatechualdehyde in Danshen from Dabie Mountains, Hubei and Mount Tai, Shandong are relatively high, 939.8 μg/g and 897.8 μg/g respectively. The content of tanshinone ⅡA in Salvia bulleyana meets the standards specified in the Pharmacopoeia of the People's Republic of China, and can be used as a source material for tanshinone ⅡA. The contents of tanshinone ⅡA, salvianolic acid B and protocatechualdehyde in Salvia yunnanensis are all relatively low, so its medicinal value needs further investigation. (4) Results of determination of 10 inorganic elements: The contents of Ca, Mg, Fe, Mo, Cu, Cd and Pb in Danshen from Shengzhou, Zhejiang are the highest, 11.07 mg/g, 10.73 mg/g, 333 mg/g, 1.447 μg/g, 27.03 μg/g, 0.2610 μg/g and 2.165 μg/g respectively, showing strong enrichment ability for trace elements. Among them, the content of Cu exceeds the standard specified in the Pharmacopoeia of the People's Republic of China, and the contents of harmful elements Cd and Pb are also relatively high, making it unsuitable for promotion as mainstream Danshen medicinal materials in the market; the contents of Cu and Pb in Salvia yunnanensis are second only to those in Danshen from Zhejiang, 0.1850 μg/g and 1.103 μg/g respectively.
