Quality study of Perillae caulis formula granules based on standard decotion / 国际中医中药杂志

Objective:To establish the quality evaluation method of Perillae caulis formula granules based on the three kind of quality indexes of standard decoction. Methods:Eighteen batches of Perillae caulis were collected from different habitats according to different technical requirements, eighteen batches of standard decoction and three batches of formula granules were prepared and the paste-forming rates were calculated. The content of Caffeic acid and Rosmarinic acid were determined and calculated by Ultra High Performance Liquid Chromatography (UPLC). Then the fingerprints of standard decoction of and formula granules of Perillae caulis were established by UPLC . The similarity values of fingerprints between formula granules and standard decoction were calculated. Results:The average paste-forming rate of standard decoction was (7.16±1.97)%. The paste-forming rates of three batches of formula granules were 5.52%, 5.25% and 5.34%, respectively. The average content of Caffeic acid and Rosmarinic acid in standard decoction was (12.06±3.37)mg/g. The contents of three batches of formula granules were 5.52, 5.82, 5.77 mg/g, respectively. Seven common fingerprint peaks were identified in the fingerprints of standard decoction and formula granules, three of which were identified as Caffeic acid, N-Feruloyl Octopus amine and Rosmarinic acid by comparison of reference substance. The fingerprints similarity of Perillae caulis dispensing granules and standard decoction were 1.000, 0.995 and 0.997, respectively. Conclusions:The quality indexes of three batches of formulation granules are consistent with standard decoction. This method can provide basis for the establishment of quality standard of Perillae caulis dispensing granules.

Textual Research on Perillae in Famous Classical Formulas / 中国实验方剂学杂志

Based on the ancient literature of all dynasties， this article makes a systematic textual research on the name， origin， producing area， quality， harvesting and processing of Zisu （Perillae） in the famous classical formulas， so as to clarify the information of the drug in different historical periods and provide a reference for the development and utilization of the related formulas. The main origin of Perillae in the ancient literature was Perilla frutescens var. frutescens （purple leaf type）， followed by P. frutescens var. acuta （purple leaf type）， but not Baisu. Modern chemical composition studies also show that there are obvious differences between Perillae and Baisu， which provides a scientific basis for distinguishing them. Although they are often treated as a species in plant classification， P. frutescens var. frutescens （purple leaf type） is recommended in the development of famous classical formulas， and Baisu should be avoided. Perillae is widely distributed， but its producing area did not record in most of the literature in the past dynasties， or the producing area is described as everywhere today. In the period of the Southern and Northern dynasties， the medicinal parts of Perillae included stems， leaves and seeds， and doctors in the Ming dynasty began to pay attention to the differentiation of different medicinal parts. The harvesting and processing methods of Perillae in the past dynasties are close to that of today. Perillae Fructus is mostly stir-fried and ground into medicine， Perillae Folium and Perillae Caulis are mainly simple cleansing. In production， we can refer to the 2020 edition of Chinese Pharmacopoeia.

Chemotype Study of Volatile Components from Perillae Folium of Different Germplasms / 中国药房

OBJECTIVE：To analyze the chemotypes of volatile components from Perillae Folium of different germplasms ，and to investigate the relationship of germplasm and leaf color with chemotype. METHODS ：The fingerprints of volatile components from 30 batches of Perillae Folium were prepared by GC-MS with P 4 peak as reference. Similarity Evaluation System for TCM Chromatographic Fingerprint （2004A edition ）was applied to evaluate the similarity and confirm common peaks. The volatile components of Perillae Folium were determined by the same GC-MS method. Qualitative Navigator （B.08.00）software was used to analyze and compare with NIST 17.0 standard mass spectrum database. The compounds corresponding to the peak were analyzed ； clustering analysis was carried out with Origin 2018 software. RESULTS ：There were 13 common peaks in the fingerprints of volatile components from 30 batches of Perillae Folium . The similarities were 0.13-1.00. Totally 54 components were identified from 30 batches of Perillae Folium of different germplasm. Cluster analysis showed that 30 batches of Perillae Folium samples could be clustered into three categories ；among them ，SCY-1，YNT-9，YNX-17，YN-28 were clustered into one category ，with phenylpropanoid-elemicin（PP-e as ）the main volatile component ，being PP-e type ；GS-4，GS-7，GS-11，GS-19，HBA-14， HBA-20，GZZ-8，LN-39，GSL-27，GSQ-32，GSQ-33，GST-31，YNW-12，LN-38 were clustered into one category ，and the content of perilla ketone （PK）in them was the highest except for LN- 38， being PK type [the content of phenylpropanoid-apiol（PP-a）in LN- 38 was higher than that of perilla ketone ，being PP-a type] ；HBS-2，HBS-3，HBS-6， C201859）HBS-15，HBS-16，HBS-24，HBS-25，GX-26，SXS-30，SCC- 36，RB-37，SC-29 were clustered into one category ，and thecontent of perillaldehyde （PA）was the highest ，being PA type.The color characteristics of Perillae Folium of different germplasm showed that Perilla frutescens （L.） Britt. var.frutescens with green leaves on both sides was PK type ，while P. frutescens （L.）Britt. var. arguta with purple leaves on one or both sides was PA type ，and P. frutescens （L.） Britt var. auriculato-dentata C. Y. Wu et Hsuan ex H. W. Li was PP-e type. CONCLUSIONS：The chemotype of volatile components in Perillae Folium have a certain corresponding relationship with their leaf colors. Most of P. frutescens （L.）Britt. var. arguta with purple leaves on one side or both sides are PA type. P. frutescens （L.） Britt. var. acuta （Thunb.）Kudo，P. frutescens （L.）Britt var. auriculato-dentata C. Y. Wu et Hsuan ex H. W. Li and P. frutescens （L.）Britt. var. frutescens with green leaves on both sides do not belong to PA type ，among which P. frutescens （L.）Britt var. frutescens is PK type ，while P. frutescens （L.）Britt var. auriculato-dentata C. Y. Wu et Hsuan ex H. W. Li is mostly PP-e type.

Quality standard of Perillae Folium based on multicomponent determination with HPLC method / 中国中药杂志

This research established the HPLC methods for the determination of perillaketone, perillaldehyde, caffeic acid, scutellarin, and rosmarinic acid in 33 batches of Perillae Folium. Kromasil C_(18)(4.6 × 250 mm, 5 μm) chromatographic column was used, and the mobile phase for determination of the perillaketone and perillaldehyde was methanol-water(55∶45) solution, at a flow rate of 1.0 mL·min~(-1), with the column temperature at 30 ℃. The mobile phase for the determination of caffeic acid, scutellarin and rosmarinic acid was methanol(A)-0.2% phosphoric acid aqueous solution(B) with gradient elution(0-20 min, 25%-30% A; 20-60 min, 30%-43% A). The flow rate was 1.0 mL·min~(-1) and the column temperature was set at 30 ℃. The results showed that the established method can achieve good separation of the five components in samples, with a good linear relationship and high accuracy, indicating that the methods can be used for the determination of Perillae Folium. The results showed that all samples contained five components. And the content of rosmarinic acid(0.04%-1.57%) > scutellarin(0.03%-0.77%) > perillaldehyde(0.02%-0.66%) > perillaketone(0.03%-0.30%) > caffeic acid(0.006%-0.07%). Thirty-three Batches of Perillae Folium can be grouped into 5 categories. There are certain content rules and region specificities under different clusters. Perillaketone, perillaldehyde, and rosmarinic acid can be used as the main markers to evaluate the quality of Perillae Folium.

Advances on Chemical Constituents of Essential Oils from Perillae Folium and Their Pharmacological Effect：A Review / 中国实验方剂学杂志

Perilla frutescens is a traditional medicinal and edible plant widely distributed in China and enjoys an extensive usage. P. frutescens contains multiple essential oils， which are composed of monoterpenes， sesquiterpenes， and their oxygen-containing derivatives. Compared with other parts of P. frutescens， Perillae Folium produce more oils， with volatile oils as the main constituents. There are many active substances in the volatile oils from Perillae Folium， mainly including perillaldehyde， perillaketone， perillaalcohol， D-limonene， β-caryophylene， etc. Such factors as germplasm， growth environment， extraction method， cultivation time， and harvest period all can trigger changes in volatile oil constituents and content from Perillae Folium. The volatile oils from Perillae Folium have diverse pharmacological effects like anti-oxidation， anti-bacteria， anti-inflammation， vasodilation， anti-tumor， and anti-depression， implying its high clinical application value. However， the chemical constituents in volatile oils from Perillae Folium are complex and unstable and their pharmacological activities are affected by many factors， so the safety and effectiveness of clinical medication fail to be guaranteed， which may has impeded the rational and effective use of these volatile oils. Many scholars in China and abroad have conducted a lot of research on the volatile oils from Perillae Folium， but there is currently no systematic and comprehensive research report on the chemical constituents of volatile oils from Perillae Folium and their pharmacological effects. This paper reviewed the relevant domestic and foreign literature， analyzed the development status of volatile oils from Perillae Folium， and summarized their extraction process， chemical constituents， and pharmacological actions， aiming to provide a reference for their further development， clinical application， and risk assessment.

Protective Effect and Mechanism of Aqueous Extract of Perillae Folium on Adriamycin-induced Oxidative Injury in HK-2 Cell / 中国实验方剂学杂志

Objective: To investigate the protective effect of Perillae Folium with aqueous extract (PFAE) on some key factors of Adriamycin (ADR)-induced oxidative injury in human renal tubular epithelial cells(HK-2), including the survival rate, oxidative injury indexes and cell apoptosis,in order to define the underlying mechanism. Method: A model of ADR-induced HK-2 cells oxidative injury was established in vitro, then cell viability was detected by cell counting kit-8 (CCK-8) after intervention with positive reference N-acetylcysteine (NAC) or PFAE (5,15,45 g·L-1) at different concentrations. According to the morphological changes under microscopy, the optimum concentration of PFAE was screened out for the follow-up experiments. Then, the experiments were divided into six groups:blank group, ADR (0.05 g·L-1) group, PFAE (15 g·L-1) group, ADR+PFAE (0.05+15) g·L-1 group, NAC (0.81 g·L-1) group, and ADR+NAC (0.05+81) g·L-1 group. After that, malondialdehyde (MDA), superoxide dismutase (SOD), total antioxidant capacity(TAC) were measured in the cell homogenate after 24 h administration. The level of reactive oxygen species (ROS) was detected by 2',7'-dichloroflurescin diacetate (DCFH-DA) fluorescence probe. Flow cytometry and TdT-mediated dUTP Nick-End Labeling (TUNEL) were used to monitor the cell apoptosis. Western blot was used to observed the expressions of mitochondrial apoptosis-associated proteins, like B lymphocyte tumor-2 gene (Bcl-2), Bcl-2 related X protein (Bax), cysteine aspartate protease-9 (Caspase-9), cysteine aspartate protease-3 (Caspase-3) and poly ADP-ribose polymerase (PARP), as well as their shear bodies. In addition, the phosphorylation protein expressions of p38 mitogen-activated protein kinase (p38 MAPK), extracellular signal-regulated kinase (ERK), c-Jun amino-terminal kinase (JNK) in mitogen-activated protein kinase (MAPK) signaling transduction pathway were detected by Western blot. Result: Compared with blank group, ADR group showed a decreased cell viability (PPPPPPPP-1. The ATC and SOD levels were increased in ADR+PFAE group and ADR+NAC group (PPConclusion: PFAE could alleviate the oxidative injury of HK-2 cells induced by ADR, and have an antioxidant effect, which inhibited cell apoptosis through mitochondrial apoptotic pathway and ERK/p38 MAPK signaling pathway.

Research progress in chemical constituents, anti-inflammatory effect and mechanism of Folium Perillae / 中国药理学与毒理学杂志

Folium Perillae (FP) is a traditional Chinese materia medica, which has been used for treating inflammatory diseases. In order to clarify the material basis of FP' s pharmacological activity and anti-inflammatory mechanism, we presented a review about some of the primary chemical components in FP, such as volatile oils, flavonoids, anthocyanins, phenolic acids, glycosides, triterpenes and steroids, and about the anti-inflammatory activity of FP extract and its major mechanism, such as regulating the viability and function of innate immune cells, controlling the balance of helper T cells, based on related research in recent years. This study aims to provide reference for further research and development of new drugs based on FP.

Ethanol Extract of Perillae Herba Enhances Pentobarbital-Induced Sleep and Non-Rapid Eye Movement (NREM) Sleep through GABA(A)-ergic Systems

Perillae Herba has been traditionally used for the sedation in the oriental countries. Therefore, this study was conducted to determine whether Perillae Herba ethanol extract (PHEE) enhances pentobarbital-induced sleeping behaviors in animals. In addition, the possible mechanisms are demonstrated. PHEE (12.5, 25 and 50 mg/kg. p.o.) reduced the locomotor activity in mice. PHEE reduced sleep latency and augmented the total sleep time in pentobarbital (42 mg/kg, i.p.)-induced sleep in mice. Furthermore, the number of sleeping mice treated with sub-hypnotic pentobarbital (28 mg/kg, i.p.) increased. PHEE (50 mg/kg. p.o.) decreased the sleep/wake cycles and wakefulness, and increased total sleeping time and NREM sleep in electroencephalogram (EEG) of rats. In addition, PHEE (0.1, 1.0 and 10 µg/ml) increased the intracellular Cl⁻ level through the GABA receptors in the hypothalamus of rats. Moreover, the protein of glutamate decarboxylase (GAD) was overexpressed by PFEE. It was found that PHEE enhanced pentobarbital-induced sleeping behaviors through GABA(A)-ergic transmissions.

Morphological characteristics of Chinese Perilla fruits / 中国中药杂志

The morphological traits of 55 Chinese Perilla fruit samples (size, 100 grains weight, color, hardness, surface ridge height) are described and the statistically analyzed. It can be divided into 6 categories by cluster analysis, namely: Ⅰ, big grain (diameter 1.5 mm above and 100 grains weight above 0.16 g), low ridge, hard; Ⅱ, big grain, low ridge, soft; Ⅲ, big grain, high ridge, soft, fruit; Ⅳ, big grain. high ridge, gray brown or dark brown; Ⅴ, small grain (diameter 1.5 mm below and 100 grain weight 0.16 g below), low ridge, hard, dark brown; Ⅵ, small grain, low ridge, hard, yellow brown. The 38 fruit samples were planted, among which 31 ones were P. frutescens var. frutescens, 4 ones P. frutescens var. crispa and 3 ones P. frutescens var. acuta. By chemotype classification, they were 29 PK type, 3 PA type, 2 PL type, 2 PP type, 1 EK type and 1 PAPK type. According the description of herb Perillae Fructus in China Pharmacopoeia, the plant originates from P. frutescens var. frutescens. In contrast, not all fruits of P. frutescens var. frutescens have accord features. The fruits with white pericarp are mainly from P. frutescens var. frutescens with purple leaves. The materials with small grain, low ridge, hard, yellow brown or dark brown, are likely to be PA type and mainly P. frutescens var. crispa.

Optimization of extraction technology for Xiaochuan Decoction based on information entropy theory / 中草药

Objective: To optimize the extraction technology of Saposhnikoviae Radix, Perillae Folium, Magnoliae Flos, Armeniacae Amarum Semen, and honey-fried Ephedrae Herba in Xiaochuan Decoction by information entropy theory. Methods: With the contents of prem-O-glucosylcimifugin, 4'-O-beta-glucopyranosyl-5-O-methylvisamminol, amygdalin, and the yield of extract as comprehensive evaluation indexes in order to optimize the extraction process parameters of orthogonal test, the weight coefficient of each index was determined by the information entropy weight method. Results: Optimum extraction technology was as follows: reflux extraction for 3 times with 10 fold water, for 1.5 h each time. Conclusion: The optimized method is stable and reliable, and can provide the reference for further development and utilization of the formula.

Optimization of Clarification and Purification Technology of Perillae folium Extract / 中国药房

OBJECTIVE:To optimize the clarification and purification technology of Perillae folium extract. METHODS:The effects of 3 clarification and purification methods as chitosan flocculation clarification,ZTC 1+1-Ⅱflocculation clarification,water precipitation on retention rate of total flavonoids and removal rate of solid of Perillae folium extract were compared to screen suit-able clarification and purification technology. With the retention rate of total flavonoids and removal rate of solid as comprehensive evaluation index,single factor and orthogonal test were designed to investigate the optimal value of concentration proportion,the amount of the flocculant,flocculation temperature and whisking speed in optimal clarification and purification method. RESULTS:Among 3 methods,the chitosan flocculation clarification was the best with concentration proportion of 1∶4,chitosan of 1.0 g/L, flocculation temperature at 60 ℃,whisking speed of 100 r/min,whisking time of 4 min,standing time of 12 h. Under the condi-tion of optimal processing,the retention rate of total flavonoids was (85.1 ± 0.75)%,and the removal rate of solid was (24.6 ± 1.33)%(n=5). CONCLUSIONS:Chitosan flocculation can be used to effectively remove the impurity of Perillae folium extract, and optimized clarification and purification technology is stable and feasible.

Anti-Oxidative and Anti-Inflammatory Effect of Water Extract from Perillae semen in RBL-2H3 Cells

The present study was to investigate anti-oxidative and anti-inflammatory activity of Perillae semen in RBL-2H3 basophilic leukemia cells. Inhibitory effect of Perillae semen onto free radical generation was determined by measuring DPPH and hydroxyl radical scavenging activities in vitro. Anti-inflammatory actions of Perillae semen extracts (100, 250, 500 microgram/mL) were assessed by testing their effects on the degranulation of mast cells. For this, beta-hexosaminidase released from RBL-2H3 cells was used and proinflammatory cytokines were measured by an ELISA kit. Our results indicated that Perillae semen water extracts effectively inhibited free radical generation. At the concentration of 500 microgram/mL of water extract, the degranulation of RBL-2H3 cells were inhibited by 42.1%. The IgE-antigen complex increased the accumulation of IL-4 and TNF-alpha secretion in RBL-2H3 cells and treatments with 250 and 500 microgram/mL of Perillae semen extracts suppressed the IgE induced secretion of IL-4 and TNF-alpha protein by 20.5, 26.9% and 14.5, 16.5% respectively. We observed that Perillae semen water extract reduced beta-hexosaminidase, IL-4, and TNF-alpha secretion in RBL-2H3 cells. These results provide that Perillae semen may be beneficial in the treatment of allergic inflammatory disease.