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

In this paper， the name， classification， origin and other aspects of Schizonepetae Herba in the famous classical formulas were researched by referring to the related herbal literature， medical books and prescription books in the past dynasties. The results showed that Schizonepetae Herba first appeared in Shennong Bencaojing （《神农本草经》） as Jiasu， while Jingjie first appeared in Wupu Bencao （《吴普本草》）， and the name of Jingjie was mainly used as the rectification of name in later generations. The name of Jiasu is mostly derived from its smell， and the name of Jingjie is mostly derived from its pronunciation. Schizonepeta tenuifolia has been highly praised in the past as a original material， and its genuine producing area is Jiangsu， Hebei and other places， medicinal part is whole herb with spike. In modern times， the quality of Schizonepetae Herba is best described as having thin stems， green spike， and aroma. In clinical application， the raw products of Schizonepetae Herba is mainly used， and the carbonisata is mainly used for hemostasis. Famous classical formulas of Huaihuasan and Danggui Yinzi are all made of Schizonepetae Spica， so it is recommended to use the dried panicle of S. tenuifolia. In Liangxue Dihuangtang， Schizonepetae Herba Carbonisata is used， therefore， it is suggested to adopt the processing method of Schizonepetae Herba Carbonisata in the 2020 edition of Chinese Pharmacopoeia.

Textual research of Schizonepetae Herba and Schizonepetae Spica / 中国中药杂志

Schizonepetae Herba and Schizonepetae Spica are well-known Chinese herbal medicines for wind dispersing and exterior releasing. Through textual research on Schizonepetae Herba and Schizonepetae Spica, the discrimination of their medicinal parts in history was clarified, and the processing, the property(nature and flavor), meridian tropism, functions, indications, usage, dosage, and the selection of decoction pieces were compared to provide the basis for clinical application. As a result, the whole herb of Schizonepeta tenuifolia was used as medicine in the early records. The aerial part and the dried spike of S. tenuifolia were used as medicines separately in the Song Dynasty, which was recorded in the Atlas of Materia Medica(Ben Cao Tu Jing). Some ancient classics emphasized that only the dried spike could be used as medicine. The separation of Schizonepetae Herba and Schizonepetae Spica meets the different needs of clinical medication and supports the concept of rational development and utilization of Chinese medicine resources. About ten processing methods for Schizonepetae Herba and Schizonepetae Spica have been recorded since ancient times, and raw and charred drugs were the major products. Raw Schizonepetae Herba is required to be used in sections, whereas raw Schizonepetae Spica in clean preparation. Both charred products should avoid scorching. Schizonepetae Herba and Schizonepetae Spica are similar in the property(pungent, bitter, and warm), meridian tropism(lung and liver meridians, as well as qi and blood aspects), and functions(releasing exterior, dispersing wind, regulating and stopping blood, promoting eruption, dispelling sores, promoting digestion, eliminating alcohol effect, etc.), but Schizonepetae Spica is superior in efficacy. For Schizonepetae Herba and Schizonepetae Spica in traditional Chinese medicinal prescriptions, the raw and charred products are similar in usage and dosage, while their focuses in clinical compatibility vary. The raw and charred products of Schizonepetae Herba and Schizonepetae Spica are widely applied clinically. Decoction pieces of different specifications can result in different efficacies and clinical applications, so medication should be performed with caution.

Effect of Schizonepetae Herba and Saposhnikoviae Radix on expression of AQP4 and AQP8 in colonic mucosa of rats with ulcerative colitis / 中国中药杂志

The aim of this paper was to investigate the effect of Schizonepetae Herba and Saposhnikoviae Radix(wind medicine) on the expression of AQP4 and AQP8 in colonic mucosa in rats with ulcerative colitis(UC). A total of 35 healthy SD male rats were randomly divided into normal group(gavaged with normal saline), DSS model group, as well as low, middle, and high dose wind medicine groups(Schizonepeta and Saposhnikovia 1∶1, gavaged at dosages of 6, 12, and 24 g·kg~(-1)·d~(-1)), with 7 in each group. UC rat model was established by free drinking of 3% dextran sulphate sodium(DSS) solution for 10 days. At the end of the 10 th day after the treatment, mice were put to death to collect colonic mucosa. The length of colon was measured; the colonic mucosal injury index(CMDI) and pathological changes of colon were observed. ELISA method was used for measuring the content of serum IL-1, IL-8, and immunohistochemical method was used to measure AQP4, AQP8 protein expressions in colon mucosa. The expressions of AQP4, AQP8 mRNA were measured by Real-time PCR. As compared with the normal group, the length of colon tissue was significantly reduced(P<0.01), CMDI scores and pathological scores were significantly increased(P<0.01), the levels of serum IL-1 and IL-8 were significantly increased(P<0.05) in model group; the immunohistochemical results showed that the protein expressions of AQP4, AQP8 were lower; the color was light yellow or brown; AQP4, AQP8 mRNA expressions in colon mucosa were significantly decreased in model group(P<0.01). CMDI scores, pathological scores, and the levels of serum IL-1, IL-8 in high, middle, low dose wind medicine groups were obvious lower than those in the model group(P<0.01 or P<0.05); the protein expressions of AQP4, AQP8 were higher; the color was chocolate brown or dark brown; the length of colon tissue, and the expressions of AQP4, AQP8 mRNA were obvious higher in wind medicine groups(P<0.01 or P<0.05). Schizonepetae Herba and Saposhnikoviae Radix could significantly improve the symptoms and histopathology of UC model rats and accelerate the intestinal mucosal healing. The mechanism may be related with up-regulating the expression level of AQP4 and AQP8 in colonic mucosa.

Evaluation of key production processes of Schizonepetae Herba formula granules based on characteristic chromatogram and quantitative transfer relationship / 中国中药杂志

The aim of this paper was to evaluate the key production processes of Schizonepetae Herba formula granules based on the new model of combining characteristic chromatogram with quantitative transfer relationship. The rationality of production process design was evaluated by studying the intermediates in different processes of formula granules, analyzing the loss of index component pulegone in each step, and establishing the characteristic chromatogram. The content of pulegone in 10 batches of standard decoction ranged between 0.067% and 0.124%(70%-130% of the average value), and the transfer rate of pulegone was 44.58%-93.97%. After the improvement of the production process, the content of pulegone in Schizonepetae Herba formula granules was 0.093%, and the transfer rate of pulegone was 68.38%, which was consistent with the parameters range of standard decoction. This study emphasized the integrality of the research process of traditional Chinese medicine(TCM) formula granules, and provided a new idea for the quality control of TCM with content determination as the main evaluation index for a long time.

Effect of Different Compatibility of Menthae Haplocalycis Herba-Schizonepetae Herba on Influenza Virus / 中国实验方剂学杂志

Objective:To research optimum compatibility components ratio for the anti-influenza virus effect of different compatibility of Menthae Haplocalycis Herba-Schizonepetae Herba. Method:The 228 SPF grade ICR rats of half male and half female were randomly divided into 19 groups, with 12 rats in each group. In each group, the ratio of flavonoids and volatile oil was determined. ICR rats were anesthetized with ether and infected with influenza A virus A/PR/8/34 (H1N1) with A 10-fold lethal dose（LD50）50 μL nasal drip infection. Than the blank control group was treated with the same amount of normal saline nasal drip control, and the rats were given the drug by gavage on the day after virus infection, the blank group and model group were given the same amount of distilled water, and the positive control group were given Oseltamivirphosphate and compound Jinyinhua granules by gavage, and once a day for 7 days. Different compatibility of volatile oil and flavonoids in Menthae Haplocalycis Herba and Schizonepetae Herba are applied to rats infected with influenza A/PR8/34 (H1N1) virus to study the pharmacodynamic screening by lung index of influenza-infected mice and histopathological examination. And antiviral effects as well as the expression of interleukin-6（IL-6）, interleukin-1β（IL-1β）, tumor necrosis factor-α (TNF-α) is studied. Result:Lung infection injury was prepared by infecting mice with influenza A virus A/ pr8/34(H1N1). Lung mass increased at the end of the experiment in the model group, and lung index was compared with that in the blank group(P<0.01). And the experimental lung weight of oseltamivirphosphate group, compound Jinyinhua granules group, volatile oil 1-volatile oil 2 group, volatile oil 1 group and volatile oil 2 group were compared with that of the model group (P<0.05). Lung index of Oseltamivirphosphate group and volatile oil group was compared with that of model group (P<0.01).Lung index of compound Jinyinhua granules group, volatile oil 1-volatile oil 2 group, volatile oil 2-flavonoids 2 group, volatile oil 1-volatile oil 2-flavonoids 1 group, volatile oil 1-volatile oil 2-flavonoids 2 group, volatile oil 1-volatile oil 2-flavonoids 2 group, volatile oil 1-volatile oil 2-flavonoids 2 group and volatile oil 2 group were compared with the model group (P<0.05). And antiviral effect as well as the expression of IL-6, IL-1β, TNF-α is studied. The expression levels of IL-6, IL-1β, TNF-α in volatile oil 1-flavonoids 1 group, volatile oil 1-flavonoids 2 group, volatile oil 1-volatile oil 2 group, volatile oil 1-volatile oil 2-flavonoids 1 group, volatile oil 1-volatile oil 2-flavonoids 2 group, volatile oil 2 groups, flavonoids 2 groups were decreased, compared with the model group (P<0.01). Conclusion:It is found that different combinations have better biological effects and good antiviral effects. This study explores the synergies compatibility rule and mechanism of action on Menthae Haplocalycis Herba and Schizonepetae Herba pairs, which will not only develop the theory of Chinese medicine compatibility but also contribute to laying a foundation for the innovation of the influenza virus drug development.

Analysis of Chemical Constituents of Volatile Oils in Menthae Haplocalycix Herba-Schizonepetae Herba and Herbal Pairs and Comparison of Anti-bacterial Effects in Vitro / 中国实验方剂学杂志

Objective: To discuss the effect of herbal compatibility on the components of volatile oils in Menthae Haplocalycix Herba and Schizonepetae Herba,and the comparison of the anti-bacterial effects in vitro,in order to provide the experimental basis for investigating the effective material basis of their herb-pair. Method: Steam distillation was used to extract volatile oils of Menthae Haplocalycix Herba, Schizonepetae Herba,and their combination. GC-MS was employed to analyze their volatile oils,and the anti-bacterial effects in vitro were compared. Result: Totally 50 compounds were identified from the volatile oils of Menthae Haplocalycix Herba;41 compounds were identified from the volatile oils of Schizonepetae Herba;and 60 compounds were identified from the volatile oils of their herbal pairs. A total of 27 common compounds were found in the samples of three volatile oils. Totally 5 new compounds were detected in the volatile oils of their herbal pairs,and 9 compounds were not detected. In vitro,volatile oils of both their herbal pairs and individual herbs had bacteriostatic effects on dung Enterococcus (ATCC2922),Staphylococcus aureus (ATCC2923),Escherichia coli (ATCC35218),and E. coli (ATCC25922). Among them,volatile oils of their herbal pairs had stronger bacteriostatic effects on dung enterococcus (ATCC2922) and staphylococcus aureus (ATCC2923) than volatile oils of individual herbs. Moreover,the order of the inhibitory effects of volatile oils on E. coli (ATCC35218 and ATCC25922) were "herbal pairs=peppermint > herba schizonepetae". Conclusion: There were some changes between the volatile oils of individual herbs and the volatile oils of their herbal pairs. These phenomena may be caused by the occurrence of certain chemical reactions and physical changes in compatibility of decocting process. The antibacterial effects of volatile oils of their herbal pairs were better than those of individual herbs in vitro.

Network pharmacological study of Schizonepetae Herba and Saposhnikoviae Radix in treatment of ulcerative colitis / 中国中药杂志

The aim of this paper was to screen the active targets of Schizonepetae Herba and Saposhnikoviae Radix in the treatment of ulcerative colitis by means of network pharmacology,and to investigate their mechanism of action. The effective components of Schizonepetae Herba and Saposhnikoviae Radix were screened out by traditional Chinese medicine systematic pharmacological( TCMSP)database,with oral bioavilability( OB) ≥30% and drug-like( DL) ≥18% selected as the thresholds. Target PPI network was built between the main components and their corresponding targets. One hundred and eighty-two human genes corresponding to the medicine target sites were obtained from Uniprot database; 3 874 genes corresponding to ulcerative colitis were obtained from Genecard database.A total of 115 intersection genes were screened from disease genes and medicine genes,and the PPI interaction analysis was conducted by using String tool. Disease-target PPI network was drawn by using Cytoscape software,and component-target-disease network was constructed. One hundred and eight nodes and 1 882 connections were found,and then Cytoscape software was used to merge the networks and filter the core network for gene GO function analysis and KEGG pathway enrichment analysis. The mechanism of Schizonepetae Herba and Saposhnikoviae Radix was then verified by animal experiment. Gene GO functional analysis suggested that biological process,molecular functions and cell components were involved,and it was found that ulcerative colitis might be related to transcription factor activity,and cytokine receptor binding,etc. Gene KEGG pathway enrichment analysis showed that the mechanism of ulcerative colitis might be associated with TNF and Toll-like receptors( TLRs) signaling pathway-mediated cytoinflammatory factors interleukin-1( IL-1) and interleukin-6( IL6). The possible mechanism of the effective components of Schizonepetae Herba and Saposhnikoviae Radix in treating ulcerative colitis might be related to intervening the cytokine receptor binding of TNF and TLRs signaling pathways,reducing the transcription of nuclear factor-kappaB( NF-κB),and inhibiting the secretion of intestinal inflammatory factors IL-1 and IL-6.

Effects of different extraction methods on composition and antibacterial activity of volatile oil from Forsythiae Fructus, Schinzonepetae Herba, and Menthae Haplocalycis Herba / 中草药

Objective To compare the chemical composition and antibacterial activity of the volatile oils from Forsythiae Fructus (FF), Schizonepetae Herba (SH), Menthae Haplocalycis Herba (MH), and their compatibility, and to explore the influence of different extraction methods on components and antibacterial activity of volatile oil. Methods GC-MS was employed to analyze the composition of the volatile oil from FF, SH, and MH extracted separately, and their mixture extracted, as well as physical mixing volatile oil of FF, SH, and MH extracted separately. The diameter of the inhibition zone and the minimum inhibitory concentration (MIC) were measured using the paper agar diffusion method and micro dilution method. The antibacterial activities of volatile oil on four common pathogens of respiratory tract were evaluated. Results When FF, SH, and MH were mixed and extracted, the main obtained components and contents of volatile oil changed. Compared to separately extracted volatile oil, eight new components emerged, seven components from FF and seven components from SH disappeared in FF and SH mixture extracted volatile oil. In FF and MH mixture extracted volatile oil, compared with separately extracted, six components from FF and eight components from MH disappeared, and eight new components emerged. As to SH and MH mixture extracted volatile oil, four components from SH and seven components from MH disappeared, and seven new components emerged. In the three herbs mixture extracted volatile oil, six components from FF, four components from SH, and two components from MH disappeared, one commom component of FF and MH, two common components of SH and MH disappeared, however, nine new components emerged. The content of pulegone significantly decreased in the three herbs mixture extracted volatile oil compared with that in separately extracted. The antibacterial experiments showed that the antibacterial effects of the volatile oils obtained by different extraction methods were different. The antibacterial effects of volatile oil extracted separately and their physical mixing volatile oil on Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, and Candida albicans were better than that from mixture extracted. Conclusion Volatile oil is an important effective component of Chinese patent medicines with relieving exterior syndrome function. Different extraction methods have a certain influence on the yield and composition of volatile oil, which ultimately affects the efficacy of drugs. Therefore, we should pay more attention to extraction method and process of volatile oil in these prescriptions.

Comparison between traditional processing and integration processing for Schizonepetae Herba based on chemical constituents and pharmacological effect / 中国中药杂志

The GC-MS method was adopted to determine the contents of β-myrcene, limonene, menthone, menthofuran, pulegone, β-caryophyllene, 1-octen-3-one and 3-octanone in volatile in Schizonepetae Herba processed by traditional processing and integration processing methods. The efficacies of Schizonepetae Herba with different processing methods were detected based on the inhibition of ear swelling induced by dimethylbenzene in mice. The rationality of the integration processing was expounded based on the comparison of chemical constituents and their pharmacological effects. The results showed that the contents of the eight chemical components in the products processed with the integrated processing method were higher than those processed with the other method. And both of the processing methods could reduce the degree of swelling and the content of TNF-α/IL-1β/IL-6 in mice serum. However, the anti-inflammatory efficacy of the products processed with the integration processing method was superior to that processed with the other method. Compared with the traditional processing method, the integration processing method ensures the quality of decoction pieces, with lower time and labor costs and higher efficiency.

Study on purification technology of total flavonoids from Schizonepetae Herba with resistance of Caco-2 cells based on grey correlation degree analysis / 中草药

Objective: Through the grey correlation analysis technology and the effect of Caco-2 colon cancer cell proliferation to evaluate the optimization of purification process in total flavonoids of Schizonepetae Herba by macroporous adsorption resin. Methods: By static adsorption and desorption rate of total flavonoids as examining indexes to screen the ratios of macroporous adsorption resin, using single factor experiment to optimize its purification process conditions of total flavonoids; The Caco-2 cell inhibition rate was determined by MTT method; At the same time, using the grey correlation degree analysis technology, the correlation between the purity of total flavonoids and proliferation inhibition rate of the tumor cell was calculated. Results: HPD-400 macroporous adsorption resin has the good adsorption separation performance of the total flavonoids from Schizonepetae Herba, Schizonepetae Herba at the concentration of 0.1 g/mL, 0.8 g/mL (medicine/wet resin) on the amount of column, using 2 BV water to remove impurity, eluting by 9 BV 70% ethanol and all of the flow rate are 4 BV/h, collecting the eluent and dry it; With the same method to do the secondary purification, the purity of total flavonoids is 92.15%, the inhibitory rate of Caco-2 cell proliferation is 86.17%, the greycorrelation coefficient of flavonoids purity and inhibition is 0.9502. Conclusion: This optimization process and the conditions are feasible and stable, the inhibitory efficacy of the in vitro Caco-2 cells is clearly. It lays the foundation of the new anti-cancer drugs based on the purification of total flavonoids in Schizonepetae Herba.