1.Differential Analysis on Chemical Composition and Pharmacodynamic Effect Between Combined Decoction and Single Decoction of Famous Classical Formula Huaganjian
Yang WANG ; Gaoju ZHANG ; Ling LI ; Liping CHEN ; Li ZHANG ; Xiao LIU ; Yuyu ZHANG ; Yuan CUI ; Minglong LI ; Chaomei FU ; Xin YAN ; Yuxin HE ; Qin DONG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(16):199-207
ObjectiveThrough qualitatively and quantitatively analysis of the differences in chemical composition between the combined decoction and single decoction of Huaganjian and comparison of their core efficacy, to explore the rationality of the flexible clinical application of Huaganjian compound preparations and single-flavored dispensing granules. MethodsUltra performance liquid chromatography-quadrupole-electrostatic field orbitrap high resolution mass spectrometry(UPLC-Q-Exactive Orbitrap MS) was used to qualitatively analyze the combined decoction and single decoction samples of Huaganjian, and meanwhile, the contents of four index components(geniposide, paeoniflorin, hesperidin and paeonol) were quantitatively analyzed by high performance liquid chromatography(HPLC). Nonalcoholic fatty liver disease(NAFLD) rat model induced by high-fat diet was applied to compare the efficacy of combined decoction and single decoction of Huaganjian. A total of 30 male SD rats were randomly divided into the control group, model group, lovastatin group(1.8 mg·kg-1), combined decoction group(1.26 g·kg-1) and single decoction group(1.18 g·kg-1). After successful modeling, lovastatin group, combined decoction group and single decoction group were given corresponding doses of drugs by intragastric administration every day, and the control group and model group were given equal amounts of normal saline by intragastric administration, after 4 weeks of administration, the serum and liver tissues were collected, and the contents of alanine aminotransferase(ALT), aspartate aminotransferase(AST), total cholesterol(TC), triglyceride(TG), low-density lipoprotein cholesterol(LDL-C) and high-density lipoprotein cholesterol(HDL-C) in serum of rats were detected, and the liver pathological examination was carried out by hematoxylin-eosin(HE) staining and oil red O staining, so as to compare differences of their efficacy. ResultsSeventy chemical components were initially identified and attributed from the lyophilized powder of the combined decoction and single decoction samples of Huaganjian, and there was no obvious difference in composition between the two. Further quantitative analysis showed that the contents of geniposide, paeoniflorin, hesperidin and paeonol in the combined decoction samples were significantly increased when compared with those of the single decoction samples(P<0.01). The pharmacodynamic results showed that compared with the model group, both the combined and single decoction groups of Huaganjian could improve the liver index of NAFLD rats, reduce the serum levels of AST, ALT, TC, TG and LDL-C, increase the serum level of HDL-C, and ameliorate the pathological changes of liver cell steatosis and fat accumulation. However, there was no significant difference in pharmacodynamic effects between the combined decoction group and the single decoction group. ConclusionThere is no significant difference between the combined decoction and single decoction of Huaganjian in terms of chemical composition, but the contents of the four index components show significantly difference. Both of them can significantly improve the fat accumulation and liver function in NAFLD rats. This study provides a reference basis for the rational clinical application and evaluation of famous classical formula compound preparations and single-flavored dispensing granules.
2.Quality evaluation of Indigo Naturalis based on multi-component quantification combined with chemometrics
Quan SUN ; Lijuan WANG ; Ling TANG ; Jing LENG ; Chaomei FU
China Pharmacy 2023;34(8):941-945
OBJECTIVE To evaluate the quality of Indigo Naturalis, and to provide reference for the quality control of Indigo Naturalis. METHODS UPLC-MS/MS method was used to determine the contents of 6 indole alkaloids (indigo, indirubin, isatin, tryptanthrin, indole and indole-3-carboxaldehyde) in Indigo Naturalis from different origins. Cluster analysis, principal component analysis and partial least squares-discriminant analysis (PLS-DA) were used to evaluate the quality of Indigo Naturalis from different origins. RESULTS The contents of indigo, indirubin, isatin, tryptanthrin, indole and indole-3-carboxaldehyde in Indigo Naturalis from different origins were 20 320.83-26 585.01, 1 327.69-3 102.25, 141.69-894.50, 2.17-5.27, 2.14-5.93 and 1.69-4.34 μg/g, respectively. The Indigo Naturalis from different areas were clustered into two categories by cluster analysis. Samples S1, S2, S4, S6, S7, S9 and S10 were clustered into category Ⅰ, and samples S3, S5, S8, S11 and S12 were clustered into category Ⅱ. Indigo Naturalis from different origins was evaluated with 3 principal components. The results showed that category Ⅰ sample scored higher and had better quality, while category Ⅱ sample scored lower and had worse quality. PLS-DA showed that indigo, indirubin, tryptanthrin and isatin were the main substances that reflected the quality difference of Indigo Naturalis. CONCLUSIONS The quality of Indigo Naturalis from different origins is different, and the quality of Indigo Naturalis of different batches from the same area is not stable. The quality evaluation method of Indigo Naturalis established in this paper is stable and reliable, which can provide a basis for the quality control of Indigo Naturalis.
3.Metabolomics-based Analysis of Toxicity Changes of Aconiti Lateralis Radix Praeparata-Zingiberis Rhizoma in Sinitang Before and After Compatibility
Wei PENG ; Dongmei LIU ; Shu FU ; Xiao WANG ; Mingquan WU ; Pilong HE ; He TU ; Chaomei FU
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(18):126-136
ObjectiveTo explore the effect and toxicity change rule of Aconiti Lateralis Radix Praeparata(ALRP) and Zingiberis Rhizoma(ZR) before and after compatibility, and to reveal the compatibility connotation of them. MethodSixty SD rats were randomly divided into blank group, blank-ALRP group, blank-ALRP-ZR group, model group, model-ALRP group and model-ALRP-ZR group, the latter three groups were injected with adriamycin via tail vein to establish the model of heart failure, and the former three groups were injected with the same amount of physiological saline via tail vein. The effects of ALRP single decoction and ALRP-ZR mixed decoction on biochemical indexes and myocardial histopathological morphology of normal rats and model rats were compared. Metabolomics analysis was performed on rat serum samples, principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA) were used to screen the differential metabolites between groups, and the differential metabolic pathways were analyzed. Combined with network pharmacology technology, the metabolites and their associated targets and pathways related to enhancing anti-heart failure efficacy and reducing cardiotoxicity were screened before and after the compatibility of ALRP and ZR, the screened representative pathways were verified by Western blot. ResultCompared with the blank group, the model group showed significant increases in the contents of brain natriuretic peptide(BNP), creatine kinase(CK), lactate dehydrogenase(LDH) and cardiac troponin(cTn)-T(P<0.01), the blank-ALRP group showed obvious increases in CK, LDH, and cTn-T contents(P<0.05, P<0.01), while the normal-ALRP-ZR group showed a significant increase in CK content(P<0.01). Compared with the blank-ALRP group, the blank-ALRP-ZR group showed a obvious decrease in LDH content(P<0.05), and pathological sections showed that both decoctions could lead to myocardial histopathological damage in normal rats. Compared with the model group, the model-ALRP-ZR group showed obvious decreases in BNP, CK, LDH and cTn-T contents(P<0.05, P<0.01), and the model-ALRP group showed obvious decreases in BNP, LDH and cTn-T contents(P<0.05, P<0.01). Compared with the model-ALRP group, the model-ALRP-ZR group showed a significant decrease in CK content(P<0.01), and both decoctions could improve the pathological morphology of myocardial tissue in the model rats. Metabolomics results showed that ALRP single decoction and ALRP-ZR mixed decoction could recover 422 and 459 metabolites in model rats, respectively. And the metabolic disruption of ALRP-ZR mixed decoction on normal rats was weaker than that of ALRP single decoction. The results of network pharmacological association analysis showed that in the aspect of ZR enhancing the anti-heart failure efficacy of ALRP, 3 metabolites such as deoxyuridylic acid were correlated to 56 metabolites, 82 targets and 13 pathways, including calcium signaling pathway, renin secretion, renin-angiotensin system, etc. In the aspect of ZR reducing the cardiotoxicity of ALRP, 3 metabolites such as tyrosol were associated with 24 metabolites, 55 targets and 14 pathways, including adrenergic signaling in cardiomyocytes and carbon metabolism and so on. Western blot results showed that the expression of angiotensin-converting enzyme(ACE), angiotensin-converting enzyme 2(ACE2) and angiotensin Ⅱ(Ang Ⅱ) in myocardial tissues of rats from the model group was significantly elevated by comparing with the blank group(P<0.01). Compared with the model group, the model-ALRP group and the model-ALRP-ZR group showed significantly decreased expression of ACE, ACE2 and Ang Ⅱ(P<0.01). Compared with the model-ALRP group, the expression of ACE2 and AngⅡ was significantly decreased in the model-ALRP-ZR group. Compared with the blank group, the expression of extracellular signal regulated kinase(ERK), protein kinase B(Akt) and cTn-I3 was significantly elevated in the blank-ALRP group and blank-ALRP-ZR group(P<0.01). Compared with the blank-ALRP group, the blank-ALRP-ZR group showed decreased expression of ERK, Akt and cTn-I3, but there was no statistical significance. ConclusionTo a certain extent, the combination of ALRP and ZR shows synergistic relationship under pathological state, and attenuated effect of compatibility under normal physiological state, and the pharmacodynamic characteristics and compatibility relationship of ALRP and ZR are closely related to the physiological state.
4.Comparison of the Contents of 14 Inorganic Elements in Cyperus rotundus from Different Origins Based on Stoichiometry
Na XU ; Shiyu WANG ; Wenbing LI ; Chaomei FU ; Hulan CHEN ; Junrong LU
China Pharmacy 2021;32(3):289-294
OBJECTIVE:To establish the method for the content determination of inorganic elements in Cyperus rotundus ,and to compare the contents of 14 kinds of inorganic elements in C. rotundus from different producing areas ,and to provide theoretical basis for its quality control and high quality resources development . METHODS :The samples were processed by microwave digestion,and ICP-MS method was used to determine the contents of Na ,Mg,K,Ca,Mn,Fe,Ni,Cu,Zn,As,Se,Sr,Cd and Pb. SPSS 23.0 software were used for principal component analysis (PCA)and cluster analysis. RESULTS :The average contents of above 14 kinds of inorganic elements in C. rotundus from different producing areas were 168.62,753.71, 6 938.33,24.31,14.69,197.77,0.60,2.43,26.89,0.21,0.06,5.64,0.05,0.32 mg/kg,respectively. The results of PCA showed that the cumulative variance contribution rate of the first four principal components was 86.203%,which could reflect most of the information of the original data. C. rotundus from Shandong ,Jiangxi,Shanxi,Hubei and Yunnan ranked the top five places in terms of comprehensive score of inorganic element contents. The results of cluster analysis showed that the samples from 9 producing areas were clustered into 5 categories,showing the characteristics of clustering by producing area. From the perspective of inorganic elements ,the quality of C. rotundus from East China ,Central China ,North China and Southwest China was better than that from South China. CONCLUSIONS :Essential trace elements like Na ,Mg,K,Ca,Mn,Fe,Cu,Zn,Sr are rich in C. rotundus,and there are small amounts of Ni ,As,Se,Cd,Pb elements in it. The contents difference of inorganic elements in C. rotundus from different origins may related to the geographical area it belongs to.
5.Comparative Study on the Contents of 4 Components in the Volatile Oil of Cyperus rotundus from Different Origins Based on HPLC Method and Multivariate Statistical Analysis
Na XU ; Yuzhen MOU ; Wenbing LI ; Chaomei FU ; Hulan CHEN ; Shiyu WANG ; Junrong LU
China Pharmacy 2020;31(23):2833-2840
OBJECTIVE: To estab lish the method for simultaneous determination of the contents of cyperotundone , nootkatone,α-cyperone and aristolone in the volatile oil of Cyperus rotundus ,compare the content differences of 4 components in C. rotundus samples from different origins ,and to provide reference for germplasm screening ,development and utilization of the medicinal material. METHODS :The volatile oil was extracted from 46 batches of C. rotundus from 12 origins. The contents of cypermenone,nootkatone,α-cyperone and aristolone in volatile oil were determined by HPLC. The determination was performed on Kromasil C 18 column with mobile phase consisted of methanol-water (68∶32,V/V)at the flow rate of 1.0 mL/min;the column temperature was 30 ℃;the detection wavelength was set at 242 nm;the sample size was 20 μL. Using the contents of above 4 components as evaluation indexes ,radar image analysis ,cluster thermal map analysis and principal component analysis were performed for comparing the quality of C. rotundus from different origins. RESULTS :The results of content determination methodology investigation met relevant requirements ;the total contents of 4 components in volatile oil from C. rotundus from different origins ranged from 136.986 4 to 538.832 1 mg/g,of which the total content of samples from Yunnan was the highest (the average value was 476.059 2 mg/g). Radar image analysis results showed that the whole contour in the 4 origins of Guangdong , Jiangxi,Guangxi and Yunnan was large relatively and better balanced ,among which the samples from Yunnan had the largest overall contour and the best balance. The cluster thermal map analysis results showed that the samples from 12 origins could be grouped into 2 categories,the first category was from Hubei ,Jiangxi,Yunnan,Sichuan,Guangdong,Shandong,Henan and Shaanxi;the second category was from Guangxi ,Shanxi,Anhui and Hainan ;the quality of samples from the first category were better than that of samples from the second category. The principal component analysis results showed that the cumulative contribution rate of the first three principal components was 96.1%,and the samples from 12 origins were mainly clustered into two categories ,which was consistent with the results of cluster thermal map analysis. CONCLUSIONS :Established HPLC method can be used for simultaneous determination of cypermenone ,nootkatone,α-cyperone and aristolone in volatile oil of C. rotundus from different origins. Among the samples from 12 origins,the quality of medicinal material from Yunnan is better.
6.Injectable thermo-responsive nano-hydrogel loading triptolide for the anti-breast cancer enhancement
Yaoyao LUO ; Jingjing LI ; Yichen HU ; Fei GAO ; George PAK-HENG LEUNG ; Funeng GENG ; Chaomei FU ; Jinming ZHANG
Acta Pharmaceutica Sinica B 2020;10(11):2227-2245
The clinical application of triptolide (TPL) in tumor therapy has been greatly limited by its toxicity and inefficient delivery. Herein, a localized and sustained-release thermo-sensitive hydrogel was developed for the intra-tumor administration of TPL. Based on the amphiphilic structure of poly (
7.Study on the Optimization of the Extraction Technology of Shengmaiyin Polysaccharide and Its Regulation Effects on Intest- inal Function of Spleen Deficiency Model Rats
Yu YOU ; Lin LUO ; Zhejie CHEN ; Meisi LIN ; Heying XU ; Bo XIONG ; Chaomei FU ; Tian XIE
China Pharmacy 2019;30(4):493-498
OBJECTIVE: To optimize the extraction technology of Shengmaiyin polysaccharide, and to investigate the regulation effects of Shengmaiyin and its polysaccharide on intestinal function of spleen deficiency model rats. METHODS: The contents of polysaccharide were determined by phenol-sulfuric acid method, and the extraction rate of polysaccharide was calculated. Using extraction rate of Shengmaiyin polysaccharide as investigation index, singel factor and orthogonal tests were used to optimize material-liquid ratio, extraction time, extraction temperature and extraction times of Shengmaiyin polysaccharide. Validation test was also conducted. Totally 80 male SD rats were randomly divided into blank group, model group, Shengmaiyin low-dose, medium-dose and high-dose groups (350, 700, 1 400 g/L, by crude drug), Shengmaiyin polysaccharide low-dose, medium-dose and high-dose groups (24.5, 49, 98 g/L, by crude drug), with 10 rats in each group. Except for blank group, other groups were given Rheum palmatum water decoction 10 mL/kg to induce spleen deficiency model, once a day, for consecutive 15 d. Since the 16th day, blank group and model group were given isovolumic water intragastrically, while other groups were given corresponding drugs, once a day, for consecutive 10 d. The general status of rats and body weights were recorded in each group. The serum contents of D-xylose, gastrin (GAS) and vasoactive intestinal peptide (VIP) were detected by phloroglucinol method or ELISA. RESULTS: The optimal extraction technology of Shengmaiyin polysaccharide was material-liquid ratio 1 ∶ 10(g/mL), extraction time 45 min, extraction temperature 80 ℃, extracting for 1 time. Results of validation test showed that extraction rates of the polysaccharide in 3 times were 7.43%, 7.64%, 7.80% (RSD=1.01%, n=3). After modeling, except for blank group, other groups suffered from loose stools, thin body and reduced food intake, and the body weight and serum level of D-xylose were decreased significantly compared with blank group (P<0.01). After last medication, above symptoms of administration groups were improved to different extents. Except for model group, body weight and serum contents of D-xylose in other groups were increased significantly than those before modeling or before medication (P<0.05 or P<0.01). Compared with blank group, body weight and serum content of GAS were decreased significantly in model group, while serum content of VIP was increased significantly (P<0.01). Compared with model group, body weight of Shengmaiyin medium-dose group and Shengmaiyin polysaccharide low-dose and high-dose groups, serum contents of D-xylose and GAS in Shengmaiyin medium-dose and high-dose groups and Shengmaiyin polysaccharide low-dose and medium-dose groups were increased significantly, while serum contents of VIP in Shengmaiyin groups and Shengmaiyin polysaccharide low-dose and medium-dose groups were all decreased significantly (P<0.05 or P<0.01). CONCLUSIONS: The optimized extraction technology of Shengmaiyin polysaccharide is stable and feasible. Shengmaiyin and its polysaccharide contribute to the recovery of intestinal function of spleen deficiency model rat, the effects of which may be associated with the secretion regulation of GAS and VIP.
8.A novel quantified bitterness evaluation model for traditional Chinese herbs based on an animal ethology principle.
Xue HAN ; Hong JIANG ; Li HAN ; Xi XIONG ; Yanan HE ; Chaomei FU ; Runchun XU ; Dingkun ZHANG ; Junzhi LIN ; Ming YANG
Acta Pharmaceutica Sinica B 2018;8(2):209-217
Traditional Chinese herbs (TCH) are currently gaining attention in disease prevention and health care plans. However, their general bitter taste hinders their use. Despite the development of a variety of taste evaluation methods, it is still a major challenge to establish a quantitative detection technique that is objective, authentic and sensitive. Based on the two-bottle preference test (TBP), we proposed a novel quantitative strategy using a standardized animal test and a unified quantitative benchmark. To reduce the difference of results, the methodology of TBP was optimized. The relationship between the concentration of quinine and animal preference index (PI) was obtained. Then the PI of TCH was measured through TBP, and bitterness results were converted into a unified numerical system using the relationship of concentration and PI. To verify the authenticity and sensitivity of quantified results, human sensory testing and electronic tongue testing were applied. The quantified results showed a good discrimination ability. For example, the bitterness of Coptidis Rhizoma was equal to 0.0579 mg/mL quinine, and Nelumbinis Folium was equal to 0.0001 mg/mL. The validation results proved that the new assessment method for TCH was objective and reliable. In conclusion, this study provides an option for the quantification of bitterness and the evaluation of taste masking effects.
9.Prediction of the anti-inflammatory mechanisms of curcumin by module-based protein interaction network analysis.
Yanxiong GAN ; Shichao ZHENG ; Jan P A BAAK ; Silei ZHAO ; Yongfeng ZHENG ; Nini LUO ; Wan LIAO ; Chaomei FU
Acta Pharmaceutica Sinica B 2015;5(6):590-595
Curcumin, the medically active component from Curcuma longa (Turmeric), is widely used to treat inflammatory diseases. Protein interaction network (PIN) analysis was used to predict its mechanisms of molecular action. Targets of curcumin were obtained based on ChEMBL and STITCH databases. Protein-protein interactions (PPIs) were extracted from the String database. The PIN of curcumin was constructed by Cytoscape and the function modules identified by gene ontology (GO) enrichment analysis based on molecular complex detection (MCODE). A PIN of curcumin with 482 nodes and 1688 interactions was constructed, which has scale-free, small world and modular properties. Based on analysis of these function modules, the mechanism of curcumin is proposed. Two modules were found to be intimately associated with inflammation. With function modules analysis, the anti-inflammatory effects of curcumin were related to SMAD, ERG and mediation by the TLR family. TLR9 may be a potential target of curcumin to treat inflammation.
10.Study on Strengthening Seedlings and Roots of Dendranthema morifolium (Ramat) Tzvel. cv. Gongju
Luhua LI ; Chaomei PAN ; Jiaxian SU ; Jiaying ZHANG ; Shuying FU
Journal of Guangzhou University of Traditional Chinese Medicine 2015;(3):494-498
Objective To explore the effect of different factors on strengthening the tissue culture seedlings and roots of Dendranthema morifolium ( Ramat) Tzvel. cv. Gongju, so as to provide experimental basis for establishing an in-vitro rapid propagation system of Dendranthema morifolium. Methods The experiment took Dendranthema morifolium ( Ramat) Tzvel. cv. Gongju tissue culture seedling as the studying object. By using tissue culture technology, different additives were added into the MS medium. Results In the seedling-strengthening culture medium, 150 mL/L of macro-element was beneficial to the growth of tissue culture seedlings of Dendranthema morifolium ( Ramat) Tzvel. cv. Gongju. The culture medium with 100 g/L of coconut milk added could obviously promote the growth of Dendranthema morifolium ( Ramat) Tzvel. cv. Gongju tissue culture seedlings. And activated carbon added to the rooting culture medium could induce the formation of tissue culture root system of Dendranthema morifolium ( Ramat) Tzvel. cv. The addition of α-naphthaleneacetic acid ( NAA) and indolebutyric acid ( IBA) at certain concentrations could promote the amount and the growth of the roots. Conclusion The tissue culture seedlings of Dendranthema morifolium ( Ramat) Tzvel. cv. Gongju are boosted by increasing the amount of macro-element, reducing the concentration of agar in the culture medium and adding coconut milk and growth hormones to the culture medium. The optimal rooting culture medium is a compound of 1/2MS, NAA 0.05 mg/L and IBA 0.05 mg/L.

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