Lithospermum erythrorhyzon has been used as a red fooddye and traditional Chinese medicine to treat wounds, skin diseases and burns. Platelet activation plays an important role in thrombosis and haemostasis. Here, we studied the inhibition of platelet activation and its active compound from the root of Lithospermum erythrorhyzon. Its ethyl acetate extract inhibited the aggregation of washed rabbit platelets induced by collagen or thrombin. Five naphthoquinone pigments, shikonin, acetylshikonin, isobutylshikonin, α-methyl-n-butylshikonin and (β,β-dimethylacrylshikonin were isolated by means of high pressure liquid chromatography. The structures were determined by comparison of their proton nuclear magnetic resonance spectra. The potency of their inhibition was in the following order : β,β-dimethylacrylshikonin≥α-methyl-n-butylshikonin$gt;isobutylshikonin$gt;acetylshikonin$gt;shikonin. It is suggested that the size of the aliphatic hydroxy group of shikonin is important for the enhancement of potency.

Background : The purpose of this study is to investigate the effect of the ridge and mulching cultivation and adapted sowing(planting)time in stable medicinal herbs production of Lithospermum erythrorhizon Siebold et Zuc. And it accomplished one’s purpose to GAP guide manual established standard method of cultivation technology, contribute to the enlargement of farm’s income. Methods and Results : Method of cultivation type with the ridge and covering material were conducted under two condition compose to open cultivation (level row : 120㎝ ridging, black polyethylene mulching) and plastic greenhouse cultivation (high row : 90㎝ ridging, non-woven fabric mulching). Sowing with Daehong varieties in April 17, transplanting July 25 by growing pot seedling for 40 days in 2015. Planting density were spaced 30㎝ apart in inter-rows 10 ㎝ apart intra-row. Fertilizing level of N-P-K-Compost (fermented mixed oil cake fertilizer) applied at 6-4-6-300 ㎏․10 a−1, 60% of basal fertilizer in March 14, 40% of top dressing were four times application in April 14 ～July 15. Plot design randomized block 3 repetition. Conclusions : Compared to level ridging on black polyethylene mulching open cultivation for growing plastic greenhouse with the high ridging non woven mulching cultivation, seed yields havesting 1.4 times, the dried roots yields increased in 0.7 times. Also, Growth characteristics by sowing time in April 16. compared to planting time in July 25. Length of stem and main root of above-ground, subterranean part growth increment rising highest. Seed and dried roots yields with increased by 71%, 30%. respectively.

Background : The purpose of this study was to big strong and root in the disease, many high quality seed of Lithospermum erythrorhizon selection by fostering, expanding the spread on a farm and Hongju subsistence production base and raw materials medicines of sources deployment for process is to contribute to increasing farm income. Methods and Results : Collected variety into the chemical mutation breeding method, Jecheon of Chungbuk, was conducted in 2001. In 2002 and 2003, two～ three individual selection in 2005 as it produces the 24 individual in 2004. From 2006 to 2010, pure line isolation as it cultivated and one foundation group. Promising in 2011, select one system. Replicated yield trial for two years beginning in 2012 a result of the stem length, providing high -quality “No. 1, Jeollanam-do” majority by assigning system name. Naju, Jangheung-gun, local excellence by implementing a local adaptability test recognized in two years. To breed a new variety named as a dwelling “ Daehong” of Lithospermum erythrorhizon the review by fostering december 2014 to be registered. Conclusions : Leaf type is the broad lancet, flowering time is new variety six days than Jecheon native variety fasted. New variety of stem length and stem thickness are check variety (87cm, 8.25mm) of 9cm tall and 0.6mm thick. Seed yields has native variety as much as 81 % compared to the many into 45.6 kg/10a. The shikonin contents of roots was 1.3 times higher compared to check variety (447 ㎍ / g). The dried root yields of the new variety are 221kg/10a a many native variety than 50%.

This study was conducted to investigate the quality of seeds, the germination rates and the days required for germination, to examine the patterns of protein expressions during the germination and to improve the techniques of managing and storing seeds and viability of the seeds of Lithospermum erythrorhizon Sieb. et Zucc. After collecting and harvesting seeds, they were classified to white and brown colors of seed coat through testing their seed size, weight, and quality. The germination rates, the days required for germination, and the protein expressions were examined with different colors of seed coats, storing temperatures and durations by treating the different plant growth regulators and primings. One hundred seed weight of white color was heavier about 1.17 g than those of brown one about 0.81 g. The germination rates in white color of seed coat was higher, 3.05 ~ 5.75%, than those in brown one. Its rates were decreased with getting longer in storage durations. There was no big differences on germination rates between storage temperatures. The plant growth regulator of GA3 and Kinetin was affected to improve the seed germination. GA3 increased the seed germination clearly at 25 ppm level, while kinetin increased it gradually from 25 to 100 ppm levels. In germination by seed primings, PEG6000 made higher germination rate with increasing their levels, whereas KNO3 increased the germination until 100 mM level and then decreased it with 200 mM unlike PEG6000. The protein expressed during the seed germination were appeared more and clearer bands in the seed after germination, especially 20 ~ 30 kDa, compared to those in the seed before germination. These results showing more and clearer bands were positively related to the germination rates which were different by seed colors, storage temperatures and durations, and plant growth regulators and primings.

This study investigates the effect of supercritical fluid extract (CMPB803-C) of Lithospermum erythrorhizon,shikonin and acetylshikonin isolated from Lithospermum erythrorhizon on IL-1β-induced chondrocytes and monosodiumiodoacetate (MIA)-induced osteoarthritis in rat. Shikonin (50μM) and acetylshikonin (3μM) treatment reduced signifi-cantly the mRNA expression and enzyme activity of matrix metalloproteinase (MMP)-1, −3 and −13 in IL-1β-inducedSW1353 chondrosarcoma cells. The chondro-protective effects of CMPB803-C and acetylshikonin were than analyzed in arat OA model using a single intra-articular injection of MIA (1㎎) in the right knee joint. CMPB803-C (200㎎/㎏) or ace-tylshikonin (5㎎/㎏) was orally administered daily for two weeks starting after 1 week of MIA injection. In the histologicalobservation, CMPB803-C and acetylshikonin clearly improved OA lesions being comparable to or better that control group.Our results demonstrated that CMPB803-C and acetylshikonin as active compound of Lithospermum erythrorhizon have astrong chondro-protective effect in OA rats, which likely attributes to its anti-inflammatory activity and inhibition ofMMPs production.

This study has assessed the anti-oxidative activities and cytotoxic effects of Lithospermum erythrorhizon ethanol extract and measured the effects of tyrosinase inhibition activities with a goal of estimating the usage of the medicinal plant as an ingredient of cosmetics. First, to perform a basic test on the extract, pH and UV-spectrum were measured. According to the measurement, the extract had control functions at pH 5.5, and maximum absorbance occurred at 530nm. In particular, DPPH (1-1-diphenyl-2-picryl-hydrazyl)-inhibiting activity (IC50) and polyphenol content were 149.81 μg/mL and 51.28±2.52 mg/mL respectively. In addition, as extract concentration increased, tyrosinase inhibition activities improved as well. In raw 264.7 cell-based MTT assay, cell survival rates were 98% at 1000 ppm and 153% at 100 ppm. Therefore, it's been confirmed that there is almost no cytotoxin. According to the test results above, it appears that the Lithospermum erythrorhizon ethanol extract would be effective in anti-oxidation and application as a cosmetic ingredient.

This study was performed to investigate the enhancement of immunomodulatory activities of Lithospermum erythrorhizon by extreme process. The extracts are WE100 (water extract for 24 hours at 100℃), WE80 (water extract for 24 hours at 80℃), EE (70% ethyl alcohol extract for 24 hours at 80℃) and EPE (extreme process for 30 minutes at 25℃, 500 MPa after 70% ethyl alcohol extracts for 3 hours at 40, 50, 60℃). Extraction yield was increased up to 5~10% by extreme process, compare to the normal extraction such as water solvent extraction, 70% ethyl alcohol solvent extraction. The cytotoxicity of the extracts was showed in the range of 12.68~15.89% at 1.0mg/ml for human lung cell (HEL299). The EPE40 was showed the lowest cytotoxicity 12.68%. The EPE60 extracted by extreme process increased the growth of human B and T cells up to 12.12×104 cells/ml and 14.88×104 cells/ml, respectively and the EPE60 greatly increased the cytokine secretion of both IL-6 and TNF-α. The extracts by extreme process also exhibited higher levels of nitric oxide production from macrophages than the lipopolysaccaharides. It can be concluded that Lithospermum erythrorhizon has immune activities and The extreme process could increase higher immune activities possibly by immunomodulatory compounds.

This study was conducted to isolate and identify the fungal pathogen causing seedling rot of Lithospermum erythrorhizon Siebold & Zuccarini, and to know the optimum growing temperature for decreasing seedling rot of Lithospermum erythrorhizon. On the basis of morphological characteristics, EF-1a sequence analysis, and pathogenecity to host plant, the fungi isolated from seedling rot and seeds of Lithospermum erythrorhizon were identified as Fusarium fujikuroi, indicating that disease causing fungus is seed-borne pathogen. Optimum temperature for germination of seeds of Lithospermum erythrorhizon was 15~20℃, but pathogenicity of Fusarium fujikuroi was shown more readily at 25~30℃. These results suggested that seedling culture of Lithospermum erythrorhizon between 15℃ and 20℃ might reduce seedling rot of Lithospermum erythrorhizon caused by seed-borne pathogen Fusarium fujikuroi.

This study was conducted to evaluate the quality variation of Lithospermum radix on the pigment contents and antioxidant activities according to different growth stages and areas of cultivation. Acetylshikonin contents showed the tendency to decrease gradually from the middle of July (0.28%) to the end of August (0.05%) and then the content was increased again to the end of October (0.25%). Shikonin content was detected as small amount of about 0.009% during the period. The weight of plants was increased from the end of September to the end of October and showed the highest value as 19.8 g on October 25. ROS scavenging activity was the highest in the early of October as IC50 value of 0.11 μg/mL. Lithospermum radix of September showed lower ROS scavenging activities than those of other growth stages as IC50 value of 1.02 and 0.49 μg/mL on September 9 and September 27, respectively. Among 17 areas cultivated Lithospermum radix, 10 areas (59%) showed 0.05-0.10% of acetylshikonin contents and 3 areas (18%) were measured to contain 0.16-0.26% of acetylshikonin.

This study was performed to enhance anticancer activities of Lithospermum erythrorhizon by eluting high amount of shikonin through ultra high pressure process. Extraction yield was increased up to 5~10% by ultra high pressure process, compare to the normal extraction processes such as water solvent extraction, 70% ethyl alcohol solvent extraction. The cytotoxicity of the extracts (1.0μg/ml) from ultra high pressure process was showed the lowest cytotoxicity 13.4% for human lung cell (HEL299). The anticancer activities showed 80~85% by adding 1.0μg/ml of the extracts from ultra high pressure process in several cancer cell lines such as AGS, Hep3B, MCF-7 and HeLa cells. Among them, MCF-7 cell of the endocrine system was highest inhibited than other cells. The anticancer activities of the extracts from ultra high pressure extraction process showed 10~15%, which was higher than the extracts from normal extraction processes. From HPLC analysis of the extracts, the contents of shikonin in the extracts from ultra high pressure process was 11.42% (w/w), which was 20% higher than others. This results indicate that ultra high pressure process could increase the extraction yield of shikonin and other contents, which resulted in higher anticancer activities.

In this study, whitening activity of Lithospermum erythrorhizon extracts were investigated according to several extraction processes: water extraction at 100℃ (WE100) and 60℃ (WE60), 70% ethyl alcohol extraction (EE) and ultra high pressure extraction (HPE) at 500 MPa for 30 minutes at 60℃. The extracts from ultra high pressure extraction showed the highest tyrosinase inhibition and melanogenesis inhibition activities as 52% and 79.5%, respectively, in adding 1mg/ml than others extraction processes. HPE extracts also showed the strong reducing power as 3.19 that absorbance at 450 mm. The contents of polyphenol in WE100, we measured as 10.1μg/ml in adding 1mg/ml. Extracts have a high total flavonoid contents by HPE as 4.1μg/ml at 1mg/ml. We can conclude that better whitening activity of extracts from high pressure extraction was due to high antioxidant activities which could be extracted by higher polyphenol and flavonoid contents in HPE than others.