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OBJECTIVE To optimize the extraction process of Sarcandra glabra. METHODS The contents of rosmarinic acid and isofraxidin in S. glabra were determined by HPLC； ultrasonic time， ultrasonic temperature， solid-liquid ratio （mL/g） and methanol volume fraction were investigated by single factor test. Based on the results of single factor test， experimental scheme was designed by Box-Behnken response surface method， and the entropy weight method was used to assign the weight of each index and calculate the comprehensive score. Taking the comprehensive score as the evaluation index， the extraction process of S. glabra was optimized， and then optimized extraction process was verified. RESULTS The optimal extraction technology of S. glabra included ultrasonic time of 40 min， ultrasonic temperature of 45 ℃， liquid-solid ratio of 50∶1， methanol volume fraction of 70%. The results of 3 times of verification experiment showed that average comprehensive score was 0.988 6， and the RSD was 0.50%. The deviation between the actual value and the predicted value （0.985 1） of each comprehensive score was within ±1%. CONCLUSIONS The optimized extraction method is stable， feasible and repeatable， which can provide reference for extraction of S. glabra.
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Sarcandra glabra, a medicinal plant in family Chloranthaceae, has been taken as an important raw material for multiple Chinese patent drugs due to its diverse indications. Considering the diversified chemical constituents and rich biological activities of S. glabra, numerous phytochemical and pharmacodynamic investigations were conducted to explore the material basis for its medicinal use. It has been found that its main chemical constituents were sesquiterpenoids, sesquiterpenoid polymers, phenolic acids, coumarins, and flavonoids. As revealed by pharmacological research, it possesses multiple biological activities like anti-inflammation, anti-bacteria, anti-tumor, anti-oxidation, and neuroprotection. Some unreported novel structures, including polymers of lindenane sesquiterpenes and monoterpenes, sesquiterpene trimers, and adducts of flavonoids and monoterpenes, have been identified from S. glabra in recent years. Moreover, biological studies relating to its anti-tumor, anti-inflammatory, and anti-oxidant activities have been deepened. This paper reviewed the chemical constituents and bioactivities of S. glabra explored over the past ten years, so as to provide a scientific basis for further development and utilization of this plant.
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Anti-Inflammatory Agents/pharmacology , Flavonoids , Phytochemicals/pharmacology , Plants, Medicinal/chemistry , SeedsABSTRACT
In this study, according to TCM theory of "liver qi stagnation forming fire", emotional stress mice model was employed to evaluate the protective effects of Qingre Xiaoyanning on herpes simplex virus type 1 (HSV-1) induced reactivation. The animal experimental protocol has been reviewed and approved by Laboratory Animal Ethics Committee of Jinan University, in compliance with the Institutional Animal Care Guidelines. BALB/c mice were divided into six groups, including mock group, HSV-1 latency group, HSV-1 reactivation group (HSV-1 latency + stress), low (0.658 g·kg-1·day-1) and high dose (1.316 g·kg-1·day-1) of Qingre Xiaoyanning groups and positive control group (acyclovir, 0.206 g·kg-1·day-1). Except for the normal group and HSV-1 latency group, all mice in other groups received a daily 12-h restraint stress for 4 days. After 7-day treatment of drugs, body weight and recurrent eye infections of mice were recorded. Brain tissues were harvested to monitor HSV-1 antigen distribution by immunohistochemical staining and detect virus titer by plaque assay. In the meantime, the mRNA and protein levels of infected cell polypeptide (ICP27) and glycoprotein B (gB) in the brain tissues were detected by RT-PCR and Western blot, respectively. The level of 4-hydroxynonenal (4-HNE) and expressions of ferroptosis-related proteins were measured by Western blot. The evaluation of malondialdehyde (MDA) content in the brain tissues was conducted by MDA assay commercial kit. The results showed that Qingre Xiaoyanning significantly retarded the decline of body weight of mice induced by HSV-1 reactivation, reduced the activation rate of HSV-1 and recurrent eye infections, declined virus titer of HSV-1, down-regulated gene and protein expressions of ICP27 and gB, and hindered the distribution of HSV-1 antigen in the brain of mice. Meanwhile, Qingre Xiaoyanning also decreased the protein expression of ferroptosis-related proteins, including DMT1, TFR1 and ALOX15 in the brain tissue of HSV-1 reactivated mice. The levels of lipid peroxidation products, 4-HNE and MDA, were also reduced by Qingre Xiaoyanning treatment. All the above results indicate that Qingre Xiaoyanning significantly inhibited HSV-1 reactivation by restraint stress, which might be related to the regulation of ferroptosis. Our findings provide a theoretical basis for the application of "clearing liver-fire" TCM on treatmenting HSV-1 reactivation-related symptoms.
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Sarglanoids A-F, six new sesquiterpenoids belonging to eudesmane (1-5) and eremophilane (6) types, were isolated from the leaves of Sarcandra glabra, a famous traditional Chinese medicine (TCM). Their structures including absolute configurations were elucidated through extensive spectroscopic analysis and electronic circular dichroism (ECD) calculations. Compounds 1-2 were rare N-containing eudesmane-type sesquiterpenoids. Compound 3 exhibited inhibitory activity against nitric oxide (NO) production in lipopolysaccharides (LPS)-induced RAW 264.7 cells with IC50 values at 20.00 ± 1.30 μmol·L-1. These findings provide scientific evidence for sesquiterpenoids as the material foundation of S. glabra.
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Molecular Structure , Plant Leaves , Polycyclic Sesquiterpenes , Seeds , Sesquiterpenes/pharmacologyABSTRACT
@#SGP-2 was an acidic polysaccharide with good hypoglycemic activity isolated from Sarcandra glabra (Thunb.) Nakai in the previous study. This study used the laser particle size analysis, transmission electron microscopy (TEM) and atomic force microscopy(AFM) analysis techniques to analyze the advanced structure of SGP-2 in the deionized water and Na2SO4 solution and discuss the structure-activity relationship between the advanced structural characterizations and the α-glucosidase inhibition activities of SGP-2 and its derivative in vitro.Results showed that SGP-2 presented aggregates and spheres in the deionized water.AFM analysis showed that the diameter of SGP-2 was 33 nm and the height was 1.84 nm, whereas compact spherical conformations with high degrees of branching were observed in 0.05 mol/L Na2SO4 solution and SGP-2 had smaller particle size in saline solution compared with that in water.SGP-2 treated by 0.5 mol/L urea and dialysis at the concentration of 1 000 μg/mL showed 98.8% inhibition activity of that from untreated SGP-2. The inhibition rate of short rod conformation with branches reached 83.3% when the temperature rose up to 140 °C, and the α-glucosidase inhibition activity was even higher than that of untreated SGP-2 under the same condition; while SGP-2 with the tangled glycan chains under the condition of carboxyl group reduced had much lower inhibition activity.Therefore, the spherical structure or the short rod conformation with branches played an important role in the activity of SGP-2. This research provides a theoretical basis for further study of structure-function relationship between the advanced structure and activity of polysaccharides.
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Objective To study the changes of inflammatory response and apoptosis in parotid gland tissues of rats after X-ray irradiation,and to explore the protective effect and possible mechanism of Sarcandra glabra on radiation-induced parotid injury in rats.Methods A total of 120 male rats were randomly divided into 5 groups (24 for each):control group,single irradiation group,radiation combined withahigh (26.8g· kg-1 · d-1),moderate (13.4g· kg-1 ·d-1) and low (6.7g· kg-1 · d-1) dosage of Sarcandra glabra group.The parotid gland of rats in the irradiation group received 15 Gy X-ray.Rats in each group were anesthetized with 2％ pentobarbital sodium (0.16 ml/100 g) at 10,40 and 70 d after irradiation and blood was collected from abdominal aorta.ROS levels in blood serum of each group were detected on the 10th,40th and 70th days after irradiation.After parotid gland tissue was taken,the pathological changes and ultrastructural changes were observed by hematoxylin-eosin (HE) staining and transmission electron microscopy,respectively.The expression level of TNF-α in parotid gland tissue was detected by immunohistochemistry,and apoptosis of parotid cells was detected by TUNEL assay.Results The content of ROS and the expression of TNF-α protein in the single irradiation group were simultaneously increased compared with the control group (t =-24.723,-35.013,-19.515,P＜ 0.05;t =-13.563,43.519,-15.249,P＜ 0.05),while they were reduced by Sarcandra glabra in a dosage dependent manner,especially in the high dosage group of Sarcandra glabra (t =5.295,8.138,6.545,P＜0.05;t =10.093,-7.868,10.539,P＜0.05).In the control group,the parotid gland tissue structure was intact,without congestion,exudation,edema,etc.For the single irradiation group,the parotid gland tissue became hyperemia,edema and inflammatory cell infiltration at 10 d after irradiation followed by fibrosis at 40 d after irradiation.These pathological alterations in the parotid gland tissue were significantly recovered when the rats were treated with Sarcandra glabra before irradiation,and the tissue damage was negatively correlated with drug dosage.TUNEL assay showed that the apoptosis rate of parotid gland cells in the single irradiation groups was higher than that in the control group (t=-4.639,-3.979,P＜0.05).Conclusions Sarcandra glabra protects parotid gland from radiation damage by scavenging radiation-induced ROS and declining inflammatory response,and thus it may be applied as a potential protective agent for radiation injury.
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Objective: To identify Sarcandra glabra and its adulterants using three DNA molecular markers including 18 S rRNA gene, ITS2 sequence and SCAR marker, and then provide the basis for its molecular authentication. Methods: 18 S rRNA gene sequence of S. glabra was obtained by PCR amplification, cloning and sequencing, and then Blast comparison was made in NCBI. The ITS2 sequence of S. glabra was obtained by PCR amplification, sequencing and annotation in ITS2 Database. In the meanwhile, the ITS2 sequences of adulterants and other plants were collected from GenBank. Using MEGA5.5, the genetic distance was calculated between species and then the ITS2 sequences were aligned to construct a phylogenetic clustering tree. SCAR molecular marker of S. glabra was obtained by RAPD. After cloning and sequencing, specific primers were designed to amplify S. glabra and its adulterants. Results: The length of 18 S rRNA obtained in our research was 1 820 bp. Blast comparison revealed that there was 99% homology between S. glabra and Chloranthaceae, which proved to be 18 S rRNA gene of S. glabra. The length of the ITS2 sequence in our research was 500 bp. Genetic distance between S.glabra and its adulterants ranged from 0.190 to 0.219, which was far more than genetic distance among adulterants (0.000-0.074). Cluster analysis showed that S. glabra and its adulterants respectively clustered into a different branch, which was far away from other plants. In our research, we obtained SCAR molecular marker of S. glabra and then a pair of specific primers were designed. Using the pair of specific primers, specific products were amplified from genomic DNA of S. glabra, but no specific products were obtained from that of its adulterants. Conclusion: We could authenticate S. glabra and its adulterants effectively with the combination of three molecular markers for establishing a novel method to identify S. glabra and its adulterants, which provides a new idea for the authentication of S. glabra.
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Objective@#To study the changes of inflammatory response and apoptosis in parotid gland tissues of rats after X-ray irradiation, and to explore the protective effect and possible mechanism of Sarcandra glabra on radiation-induced parotid injury in rats.@*Methods@#A total of 120 male rats were randomly divided into 5 groups(24 for each): control group, single irradiation group, radiation combined with a high(26.8 g·kg-1·d-1), moderate(13.4 g·kg-1·d-1) and low(6.7 g·kg-1·d-1) dosage of Sarcandra glabra group. The parotid gland of rats in the irradiation group received 15 Gy X-ray. Rats in each group were anesthetized with 2% pentobarbital sodium (0.16 ml/100 g) at 10, 40 and 70 d after irradiation and blood was collected from abdominal aorta. ROS levels in blood serum of each group were detected on the 10th, 40th and 70th days after irradiation. After parotid gland tissue was taken, the pathological changes and ultrastructural changes were observed by hematoxylin-eosin (HE) staining and transmission electron microscopy, respectively. The expression level of TNF-α in parotid gland tissue was detected by immunohistochemistry, and apoptosis of parotid cells was detected by TUNEL assay.@*Results@#The content of ROS and the expression of TNF-α protein in the single irradiation group were simultaneously increased compared with the control group (t=-24.723, -35.013, -19.515, P<0.05; t=-13.563, 43.519, -15.249, P<0.05), while they were reduced by Sarcandra glabra in a dosage dependent manner, especially in the high dosage group of Sarcandra glabra (t=5.295, 8.138, 6.545, P<0.05; t=10.093, -7.868, 10.539, P<0.05). In the control group, the parotid gland tissue structure was intact, without congestion, exudation, edema, etc. For the single irradiation group, the parotid gland tissue became hyperemia, edema and inflammatory cell infiltration at 10 d after irradiation followed by fibrosis at 40 d after irradiation. These pathological alterations in the parotid gland tissue were significantly recovered when the rats were treated with Sarcandra glabra before irradiation, and the tissue damage was negatively correlated with drug dosage. TUNEL assay showed that the apoptosis rate of parotid gland cells in the single irradiation groups was higher than that in the control group (t=-4.639, -3.979, P<0.05).@*Conclusions@#Sarcandra glabra protects parotid gland from radiation damage by scavenging radiation-induced ROS and declining inflammatory response, and thus it may be applied as a potential protective agent for radiation injury.
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OBJECTIVE: To develop an HPLC method for simultaneous determination of seven active constituents including neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, quercetin-3-O-β-D-glucuronide, isofraxidin, kaempferol-3-O-β-D-glucuronide and rosmarinic acid in different parts and various collecting time of Sarcandra glabra. METHODS: The DIKMA Diamonsil C18 (4.6 mm × 250 mm, 5 μm) column was used. acetonitrile(A) -0.2% phosphoric acid(B) was used as the mobile phase with gradient elution(0 - 10 min, 5% A→15% A; 11 - 18 min, 15% A→25% A; 18 - 25 min, 25% A; 25 - 40 min, 25% A→40% A), at a flow rate of 1.0 mL•min-1. The detection wavelength was set at 344 nm and the column temperature was maintained at 35 ℃. RESULTS: The calibration curves of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, quercetin-3-O-β-D-glucuronide, isofraxidin, kaempferol-3-O-β-D-glucuronide and rosmarinic acid were in good linearity over 9.32 - 466.00, 11.25 - 562.50, 10.94 - 547.00, 8.68 - 434.00, 10.48 - 524.00, 9.66 - 483.00 and 10.86 - 543. 00 μg•mL-1 (r = 0.999 6 - 0.999 9), respectively. The average recoveries (n = 6) were in the range of 99.0% - 101.5% and RSD were in the range of 0.85% - 1.8%. The optimal parts and collecting time of Sarcandra glabra are aerial parts and from September to November according to the comprehensive evaluation. CONCLUSION: The method is precise and reliable, can be used to determin the content of seven chemical constituents in Sarcandra glabra and provide a reference for collecting in cultivate area.
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Rosmarinic acid is one of anti-tumor ingredients in the Sarcandra glabra. After treatment with 0, 10, 30, 90, 270 and 810 μmol·L⁻¹ rosmarinic acid for 24, 48, 72 hours respectively, the inhibitory effects on MCF-7 and MDA-MB-231 cells were observed by CCK-8 and cell wound healing test. No significant inhibition effecton proliferation and migration was observed in MCF-7 cell. However, 90, 270 and 810 μmol·L⁻¹ rosmarinic acid could inhibit the proliferation and migration of MDA-MB-231 cells in a dose-dependent and time-dependent manner. Flow cytometry was further used to detect apoptosis ratios of MDA-MB-231 cells after Annexin V-FITC/PI staining, and significant apoptosis was observed after rosmarinic acid treatment. Real-time PCR and Western blot tests were carried out to detect the expressions of apoptosis-related genes. The down-regulation of the Bcl-2 expression and the up-regulation of the Bax expression were observed in MDA-MB-231 cells after rosmarinic acid treatment. The results suggested that rosmarinicacid can inhibit the proliferation and migration, and induce the apoptosis of MDA-MB-231 cells, which may be correlated with the down-regulation of Bcl-2 gene and the up-regulation of Bax gene.
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The plants of Chloranthaceae are widely distributed and rich in resources in our country, records of ancient herbs indicaded that various species of plants of Chloranthaceae can be used for medicinal purposes, especially the Sarcandra glabra which with the least toxicity and possessed the function of clearing heat and cooling blood, activating blood to eliminate spots and removing wind and dredging collaterals. Based on the theory of herbage and by the method of consulting the past herbal literature, we summarized and analyzed the medicinal history, distribution characteristics of herbage geography, characteristics of herbage ecology, standard collection situation and the modern toxicology research of Chloranthaceae plants. Therefore we explained the scientificity for selection of medicinal herbs of Chloranthaceae plants, and provided a theoretical and scientific basis for further research of Chloranthaceae plants.
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Objective: Sarcandra glabra was recognized as an important research material attributing to its high medicinal value and economic value. However, little information was known about its genomics and regulatory pathway participating in reproductive development. For the first step to understand the molecular basis and further explore genes which related to metabolism and resistance in S. glabra. Methods: A SMART full-length complementary DNA library from the leaves tissue was constructed and characterized to providing the experimental basis for discovery of functional genes of S. glabra. The assembly expressed sequence tag (EST) data were completed by ABI3730 DNA program. A high quality full-length cDNA library was constructed successfully from S. glabra leaves. Results: The titer of library was 1.14×107 pfu/mL and the average length of inserted fragments was 1 000 bp. A total of 221 clones were sequenced from the cDNA library and obtained 177 EST sequences. The EST sequences were assembled into 151 unigenes including 12 contigs and 119 singletons (79%). EST exhibited significant similarity with known putative functional nucleotide sequences in the GenBank database. These genes were mostly involved in cell development, signal transduction, protein synthesis, transcription, stress tolerance response, energy metabolism based on molecular function of GO annotation. Conclusion: This report constructs a full-length-cDNA library and analyzes the bioinformatics of the related EST sequences, and then offers a reference to genomic research of S. glabra.
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Objective To study the effect of Sarcandra Glabra on the expression of signal transduction molecules of TGF-β1/Smads signaling pathway in miniature pig of radiation-induced lung injury.Methods 75 miniature pigs were divided into control group,radiation group and radiation plus medication group randomly.At 1 week before exposure of right lung with 15 Gy γ-rays,the miniature pigs in radiation plus medication group were given Sarcandra glabra,while those in the other groups received an equal amount of saline.Right lung were taken at weeks 2,4,8,12 and 24 after irradiation,the pathological changes in the lung tissue were observed by HE staining,and the expression of mRNA and protein of TGF-β1,Smad2,Smad3,and Smad7 were detected by real-time PCR and western blotting,respectively.Results Sarcandra glabra reduced the inflammation and fibrosis of the lung tissue in miniature pig after irradiation.Compared with control group,the expressions of TGF-β1 and Smad3 were significantly increased at 2 weeks after irradiation(P ＜ 0.05),Smad2 and Smad7 were increased at 8 and 12 weeks after irradiation(P ＜ 0.05),respectively,in the radiation group.Compared with the radiation group,the expressions of TGF-β1 and Smad2 were significantly decreased(P ＜ 0.05) from the fourth and eighth week,respectively,Smad3 had no obvious change while Smad7 was significantly increased from the second week in the radiation plus medication group (P ＜ 0.05).Conclusions Sarcandra Glabra plays protective effect on radiation-induced lung injury in miniature pig by regulating TGF-β1,Smad2 and Smad7 expressions in the TGF-β1/Smads signaling pathway.
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Objective To study the genetic diversity in Fujian Sanming Sarcandra Glabra from different geographical provenances;To construct genetic relationship diagram.Methods DNA in leaves was extracted by CTAB. Analysis and evaluation of DNA molecular level of 45 samples of different geographical provenances were conducted by ISSR method. POPgen32 software was used to calculate the genetic diversity and establish gene trees. NTSYS software was used to carry out cluster analysis.Results Six ISSR primers amplified 630 bands. Genetic diversity analysis showed that the average effective number of alleles of 45 varieties was 1.620 2;the average Nei's genetic diversity index was 0.364 5;the average Shannon information index was 0.543 2. Each point had different levels of genetic diversity. Nei's genetic diversity index of the maximum value was 0.497 2, and the minimum value was 0.107 8;Maximum Shannon information index was 0.690 3, and the minimum value was 0.219 2. Cluster analysis results showed that 45 varieties and 14 loci band data were the primitive matrix. 630 genetic similarity coefficients between two different species were obtained. The maximum similarity coefficient among different groups was 1.0, and the minimum was 0.516.Conclusion Different varieties of Fujian Sanming Sarcandra Glabra exist abundant genetic variation and has the molecular basis of abundant species. Using 0.610 as the threshold value can divide Sarcandra Glabra from 45 different geographical provenances into 6 groups. The genetic distance and geographical distance was not related.
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Objective: To study the ribosomal DNA internal transcribed spacer (rDNA-ITS) sequences of Sarcandra glabra from different areas and five plants in Chloranthus Swartz, and to provide pattern recognition and thread for the species identification of S. glabra. Methods: The ITS sequences of 18 populations of S. glabra and six populations of Chloranthus spicatus were amplified by PCR with universal primer of ITS and sequenced, and the ITS sequences of the other plants in Chloranthus Swartz were searched in GeneBank. Data were analyzed by ClustalX 2.1, and a cluster analysis was presented by UPGMA. Results: The semblance of ITS sequences of S. glabra from different areas was 99%. The total mutation rate of ITS1 sequence (2.7%) was higher than that of ITS2 sequence (1.4%). However, compared with other plants in Chloranthus Swartz, the total mutation rate of ITS2 sequence (20.3%-22.7%) was higher than that of ITS1 sequence (15.9%-18.3%). The cluster analysis showed that there was little variation among the 18 populations of S. glabra, but there was significant difference with the five plants of Chloranthus Swartz. Conclusion: ITS sequence can be used to identify the plants from different areas and in different genus, ITS sequence of S. glabra has several specified information sites to identify the five plants in Chloranthus Swartz, with significantly different cluster analyseis, and is the active molecular marker for the species identification of S. glabra and plants in Chloranthus Swartz.
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Objective: To analyze the correlation of DNA fingerprints of Sarcandra glabra with different quality levels and its quality. Methods: Using ISSR-PCR, 100 ISSR primers (UBC801-UBC900) were screened, and 23 of them were polymorphic. The 23 ISSR primers were used to amplify 18 S. glabra samples from different habitats. Based on the 18 ISSR amplified bands, the data base of amplified bands fingerprint was established using Excel. Results: one hundred and ninty-eight bands were obtained. Each primer amplified eight bands on average. The number of polymorphic DNA bands was 184, and the polymorphic proportion of DNA bands was 92.9%. Seven sites in two primers (UBC811 and UBC825) screened from 23 polymorphic ISSR primers were in one group, and seven sites in three primers (UBC827, UBC834, and UBC842) were in another group. The DNA fingerprints of 18 S. glabra samples were established, and U844-6 bands were screened from 184 polymorphic bands to correlate with sample quality. Conclusion: ISSR molecular markers could identify the DNA fingerprints of 18 S. glabra samples, and screen one band that is related to the quality.
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Objective To explore the effect and underlying mechanism of Sarcandra glabra Nakai on radiation pneumonnopathy in miniature pigs irradiated with 60Co γ-rays.Methods Totally 60 miniature pigs were randomly divided into blank control group,irradiated group with 15 Gy of 60Co γ-rays,and intervention group administered with Sarcandra glabra Nakai at the dosage of 0.3 g/kg before 15 Gy irradiation.At 4,8,12 and 24 weeks post-irradiation,the respiratory rate and weight were observed,and a portion of the right lung tissue was taken out to conduct HE and Masson staining examination,while the expressions of TGF-β1 and TNF-α as well as the content of hydroxyproline (HP) were detected.Results The respiratory rate,the lung coefficient and the content of HP in the irradiated group showed an increasing trend at 4,8,12 and 24 weeks post-irradiation and significantly higher than those in other two groups (F =21.035,146.014,32.610,P ＜ 0.05).Those indicators were indistinguishable between the control group and intervention group at 4 and 8 weeks post-irradiation (F =0.055,2.456,5.581,P ＞ 0.05),while the indicators of the irradiation group were significantly higher than that of intervention group (F =91.897,93.149,83.487,P ＜0.05) at 12 and 24 weeks post-irradiation.The pulmonary histopathological results showed that no inflammatory response or fibrosis was found in the control group and intervention TGF-β1 and TNF-α in the intervention group were negatively expressed at 4 and 8 weeks post-irradiation and had low expressions at 12 and 24 weeks post-irradiation which was lower than that of the irradiated group.Conclusions Sarcandrae has protective effect against radiation-induced lung injury in miniature pigs probably by inhibiting the expressions of TNF-α and HP.
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Objective To study the effects of drought stress on chlorophyll fluorescence characteristics and energy dissipation of Sarcandra glabra and get the method of alleviating drought stress, so as to provide theoretical basis against drought in planting. Methods Several physiological indexes of S. glabra treated by 5-aminolevulinic acid (ALA) under drought stress (PEG-6000 at the concentration of 15%) were measured, such as the contents of photosynthetic pigment, chlorophyll fluorescence parameters, and energy parameters. Results Exogenous ALA obviously increased the content of chlorophyll and carotenoids, enhanced the maximum fluorescence (Fm), photochemical efficiency of photosystem II (PSII, Fv/Fm), potential photochemical efficiency (FWFo), photochemical efficiency (Fv'/Fm'), PSII actual photochemical efficiency (φPSII), photochemical quenching coefficient (qP), electronic transfer rate (ETR), and photochemistry rate (PCR), as well as significnatly decreased the level of minimum fluorescence (Fo), non-photochemical quenching (NPQ) coefficient, and heat dissipation rate (HDR). The proportion of ALA absorbed light in photochemistry (P) was increased, the fraction of antenna pigment heat dissipation (D) and excess energy (E) for NPQ was decreased. The fraction off was the main pathway for excessive energy dissipation. ALA could promote the redistribution of energy reasonablely. Conclusion Exogenous ALA (100 mg/L) could significantly reduce the dissipation of excess excitation energy, improve the photochemical electron transport efficiency, and efficiently protect leaf blade of S. glabra from PSII damage under drought stress. ALA could obviously promote the drought resistance of S. glabra plantlet.
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Microwave-assisted extraction (MAE) was used for extraction of effective components of sarcandra glabra (Thunb.), and then chromatographic fingerprint of sarcandra glabra (Thunb.) was studied by high performance liquid chromatography/diode array detector (HPLC/DAD). The conditions of MAE were optimized by an orthogonal experiment, and then the authentication and validation of the chromatographic fingerprint were conducted. Nine peaks were identified as common peaks in the fingerprint chromatograms, and isofraxidin was considered as a reference compound and quantified. Relative standard deviations of retention time and peak area of each component were less than 3% and 8%, respectively. Similarity and difference analysis were conducted by use of PCA and relation coefficient. Twenty batches of sarcandra glabra (Thunb.) samples from two different producing areas could be classified into two different groups in the PCA model. The results showed that MAE-HPLC/DAD method was simple, efficient and stable for the study of complex chromatographic fingerprint of sarcandra glabra (Thunb.), which could provide more reliable and precise information for quality evaluation.
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Microwave-assisted extraction (MAE) was used for extraction of effective components of sarcandra glabra (Thunb.), and then chromatographic fingerprint of sarcandra glabra (Thunb.) was studied by high performance liquid chromatography/diode array detector (HPLC/DAD). The conditions of MAE were optimized by an orthogonal experiment, and then the authentication and validation of the chromatographic fingerprint were conducted. Nine peaks were identified as common peaks in the fingerprint chromatograms, and isofraxidin was considered as a reference compound and quantified. Relative standard deviations of retention time and peak area of each component were less than 3 % and 8 %, respectively. Similarity and difference analysis were conducted by use of PCA and relation coefficient. Twenty batches of sarcandra glabra (Thunb.) samples from two different producing areas could be classified into two different groups in the PCA model. The results showed that MAE-HPLC/DAD method was simple, efficient and stable for the study of complex chromatographic fingerprint of sarcandra glabra (Thunb.), which could provide more reliable and precise information for quality evaluation.