1.Triterpenes from Callicarpa integerrima Champ.
Chenchen ZHU ; Li GAO ; Zhongxiang ZHAO ; Chaozhan LIN
Acta Pharmaceutica Sinica 2012;47(1):77-83
A new triterpenoid saponin and fourteen known triterpenoids were isolated from the methanol extract of the stems and leaves of Callicarpa integerrima Champ, which is used in Chinese folk medicine for stopping bleeding, expelling the wind, dissipating stagnation, and treating scrofula, by using various chromatographies, such as silica gel, Sephadex LH-20 and RP-C18 column chromatography. Their structures were identified as a new compound 2alpha, 3beta, 19alpha, 23-tetrahydroxy-olean-12-en-28-oic acid-28-O-beta-D-glucopyranosyl-(1 --> 4)-beta-D-glucopyranoside (1), together with fourteen known compounds: oleanolic acid (2), 3-acetyl oleanolic acid (3), 3beta-O-acetyl ursolic acid (4), 2alpha-hydroxy-ursolic acid (5), 2alpha, 3beta, 19alpha, 23-tetrahydroxy-urs-12-en-28-oic acid (6), alpha-amyrin-3-O-beta-D-glucopyranoside (7), pomolic acid (8), betulinic acid (9), ursolic acid (10), 2alpha, 3beta, 19alpha, 23-tetrahydroxy-olean-12-en-28-oic acid (arjungenin) (11), 2alpha-hydroxy-oleanolic acid (12), hederagenin (13), 2alpha, 19alpha-dihydroxy-ursolic acid (14) and pruvuloside A (15), by the spectroscopic techniques of NMR, HMBC, IR and MS, separately. All these compounds were obtained from this plant for the first time, and compounds 3, 4 and 15 were isolated from genus Callicarpa L. for the first time.
2.Simultaneous determination of seven flavonoids in Nervilia fordii with HPLC.
Li ZHANG ; Chenchen ZHU ; Zhongxiang ZHAO ; Chaozhan LIN
Acta Pharmaceutica Sinica 2011;46(10):1237-40
The study is to develop an HPLC method for simultaneous determination of rhamnazin (1), rhamnocitrin (2), rhamnetin (3), rhamnazin-3-O-beta-D-glucopyranoside (4), rhamnazin-3-O-beta-D-xylopyranosyl-(1-->4)-beta-D-glucopyranoside (5), rhamnazin-3-O-beta-D-glucopyranosyl-(1-->4)-beta-D-glucopyranoside (6), and rhamnocitrin-3-O-beta-D-glucopyranosyl-(1-->4)-beta-D-glucopyranoside (7) in Nervilia fordii. The separation was performed on a Kromasil C18 column (250 mm x 4.6 mm, 5 microm) with 0.4% phosphoric acid-acetonitrile as the mobile phase in a gradient elution at a flow rate of 1.0 mL x min(-1). The detect wavelength was set at 256 nm, and the column temperature was set at 40 degrees C. There were good linear relationships between the logarithm values of concentrations and those of the peak areas of seven flavonoids (1-7) in the range of 0.55-70.00 microg x mL(-1) (r = 0.9997), 0.86-110.00 microg x mL(-1) (r = 0.9997), 0.39-50.00 microg x mL(-1) (r = 0.999 7), 0.55-70.00 microg x mL(-1) (r = 0.999 5), 1.33-170.00 microg x mL(-1) (r = 0.9998), 1.33-170.00 microg x mL(-1) (r = 0.9998), 0.16-20.00 microg x mL(-1) (r = 0.9995), respectively. The recoveries of the seven flavonoids were between 97.19%-99.45%, the relative standard deviations (RSDs) were between 0.91%-2.69%. The established method is rapid, accurate with high repeatability, which could provide scientific evidence for the quality control of Nervilia fordii.
3.Study on oriented differentiation of bone marrow mesenchymal stem cells by fibroblast in rat uterine ligament with mechanical stretch
Chenchen REN ; Ruifang REN ; Bing ZHAO ; Xi ZHANG ; Yongjie JIANG
Chinese Journal of Obstetrics and Gynecology 2011;46(7):527-532
Objective To investigate the effect on the differentiation of bone marrow mesenchymal stem cells (BMSC) with non-contact co-culture with mechanical stimulated ligament fibroblasts. Methods A cyclic 10% uniaxia strain at 1 Hz was applied on rat pelvic ligament fibroblasts, then were co-cultured with BMSC for 3, 6 and 12 days in non-contact condition. The protein expression of collagen Ⅰ ,Ⅲ in BMSC were detected by SP method and revealed by the mean gray value. The mRNA expressions of collagens type Ⅰ and type Ⅲ in the BMSCs were measured with real-time (RT)-PCR ,and the results were indicated by the ratio between the mRNA and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) . Results (1) Protein expression; after 3 days co-culture with pelvic ligament fibroblasts, expression of collagen Ⅰ and Ⅲ in BMSC are 82. 4 ± 3. 4 and 76. 8 ± 2. 5. When compared with 80. 2 ± 2. 6 and 74. 6 ± 1. 1 in BMSC without co-culture, there was no significant difference (P > 0. 05) . After 6 days co-culture with pelvic ligament fibroblasts, the expression of collagen Ⅰ and Ⅲ of 126. 6 ±2. 2 and 118. 6 ± 1. 4 in BMSC were significantly higher than 82. 7 ±3. 0 and 76. 2 ± 1. 3 in BMSC without co-culture (P < 0. 05). Similarly, after 12 days co-culture with pelvic ligament fibroblasts, the expression of collagen Ⅰ and Ⅲ of 135. 3 ±3. 4 and 128. 7 ± 2. 6 in BMSC were significantly higher than 86. 6 ± 1. 3 and 81. 8 ± 1.4 in BMSC without co-culture (P <0.05). (2)mRNA expression:after 3 days co-culture with pelvic ligament fibroblasts , the mRNA expression of type Ⅰ and type Ⅲ collagens in BMSC are 2. 10 ±0. 20 and 1. 20 ±0. 30. When compared with mRNA expression of 2. 01 ±0. 12 and 1. 13 ±0.21 in BMSC without co-culture, no significant difference were observed (P > 0. 05). After 6 days co-culture with pelvic ligament fibroblasts , the mRNA expressions of type Ⅰ and type Ⅲ collagens mRNA were 5. 60 ±0. 21 and 2. 61 ±0. 20, which were significantly higher than 3. 70 ±0. 33 and 1. 82 ± 0. 14 in BMSC without coculture (P < 0. 05). After 12 days co-culture with pelvic ligament fibroblasts, the mRNA expressions of type Ⅰ and type Ⅲ collagens of 5. 91 ±0.31 and 2. 92 ±0. 23 were significantly higher than 4. 04 ±0. 21 and 2. 04 ±0. 13 in BMSC without co-culture (P <0. 05). Conclusion Non-contact co-culture with mechanical stretch stimulated ligament fibroblasts, it might promote synthesis of types Ⅰ and Ⅲ collagen in rat BMSCs and induced BMSC differentiated into pelvic ligament fibroblasts.
4.Total triterpeniods from Rabdosia lophanthoides var.gerardianus with macro porous resin
Chaozhan LIN ; Cuixian ZHANG ; Zhongxiang ZHAO ; Chenchen ZHU
Chinese Traditional Patent Medicine 1992;0(01):-
AIM: To study the enrichment-purification process of total triterpeniods from Rabdosia lophanthoides var.gerardianus with macro porous resin. METHODS: The purify of the total triterpeniods was used as marker to optimize the adsorptive capacity and elution characteristics. RESULTS: The result showed that 20 mL of the extraction solution(0.5 g/mL) was purified with a column of macro porous resin(d15 mm?h120 mm,V=20 mL,dried weight 10 g) and washed with distilled water,then eluted with 80 mL 60% ethanol and 160 mL 90% ethanol in proper order.With macro reticular resin to adsorb and purify,the elution ratio of total triterpeniods of 90% ethanol fraction was 93.2% and the purity reached above 20%. CONCLUSION: This process of applying macro reticular resin to adsorbing and purifying total triterpeniods from Herba Rabdosia lophanthoides var.gerardianus was successful.
5.Study on Antioxidant Stilbenes from the Rhizomes of Smilax china
Zhongxiang ZHAO ; Jing JIN ; Chenchen ZHU ; Cuixian ZHANG ; Jinlan RUAN
Traditional Chinese Drug Research & Clinical Pharmacology 1993;0(02):-
Objective To study the stilbene constituents from the traditional Chinese medicine Smilax china and to determine their antioxidant activity. Methods The compounds were separated and purified by column chromatography with silica gel, RP C18, and Sephadex LH- 20, and were identified by IR, MS, NMR. DPPH method was used to evaluate the free radical scavenging activity of the isolated compounds. Results Three compounds were isolated from the EtOAc fraction of the rhizomes of S. china and were identified as: resveratrol (1), oxyresveratrol (2) and 3, 5, 3′ , 4′ - tetrahydroxylstilbene (3). Compounds 1~ 3 showed strong antioxidant activity, and could scavenge DPPH free radicals, effectively. At the concentration of 50 ? mol/L, their DPPH free radical scavenging rates were 79.47 % , 89.89 % and 93.86 % , respectively. Conclusion Stilbenes might be the material foundation of antioxidant activities of rhizomes of S. china.
6.Expression of decoy receptor 3 in pancreatic cancer tissue and its clinical significance
Xinhui ZHANG ; Xin ZHAO ; Yangsui LIU ; Xiaochuan WANG ; Chenchen KONG
Chinese Journal of Hepatobiliary Surgery 2015;21(12):830-832
Objective To detect the expressions of decoy receptor 3 (DcR3) in pancreatic cancer tissues and to analyze the significance of DcR3 in the diagnosis, treatment and prognosis of patients with pancreatic cancer.Methods The expressions of DcR3 in pancreatic cancer tissues (n =100), paracancer tissues (n =15) and normal tissues (n =15) were detected with immunohistochemical method (Envision method).Results The positive rate of DcR3 in pancreatic cancer tissues was significantly higher than that in adjacent-tumor pancreatic cancer tissues (86.0% vs.46.6%, P < 0.05).The positive rate of DcR3 in adjacent-tumor pancreatic cancer tissues was significantly higher than that in normal tissues (46.6% vs.13.3%, P < 0.05).In clinical stage Ⅲ, the positive rate of DcR3 was significantly higher than that in stage Ⅱ and stage Ⅰ (100% vs.87.0%, P<0.05;100% vs.62.5%, P<0.05).There were significant differences among the three groups (P < 0.05).With lymph node metastasis, the DcR3 positive rate was significantly higher than that in the group without lymph node metastasis (93.4% vs.79.6%, P < 0.05).In poorly differentiated pancreatic cancer, the positive rate of DcR3 was significantly higher than that in the highly differentiated group (100% vs.64.0%, P <0.05), the positive rate of DcR3 was significantly higher in the moderately differentiated group than that in the highly differentiated group (88.6% vs.64.0%, P < 0.05) , There were significant differences among the three groups (P < 0.05).There was no significant difference in the positive rate of DcR3 between the different age groups or the different gender groups (P > 0.05).Conclusions The expression levels of DcR3 in patients with pancreatic cancer gradually increased from normal tissues to paracancer tissues, to pancreas tissues.The expression level of DcR3 protein was closely related to clinical stage, degree of tissue differentiation and presence of lymph node metastasis, but not associated with age, sex, and tumor diameter size.
7.Ophiopogonin D protects cardiomyocytes against doxorubicin-induced injury through suppressing endoplasmic reticulum stress.
Chen MENG ; Caihua YUAN ; Chenchen ZHANG ; Mingda WEN ; Yanhong GAO ; Xiaoyu DING ; Yingyu ZHANG ; Zhao ZHANG
Acta Pharmaceutica Sinica 2014;49(8):1117-23
This study aimed to examine whether ophiopogonin D (OP-D) is capable of protecting cardiomyocytes against DOX-induced injury and the mechanisms involved. H9c2 cells were cultured. MTT assay was used to evaluate cell viability and toxicity. Mito-tracker as fluorescence probe was used to measure ROS content raised from mitochondria. The mRNA and protein expression of ATF6alpha, GRP78 and CHOP were analyzed using real-time PCR and Western blotting, respectively. The results showed that a significant endoplasmic reticulum stress (ERS) was induced upon exposure of H9c2 cells to DOX as indicated by the increase in the expression of ERS related proteins, which was paralleled with the accumulation of reactive oxygen species (ROS) and decrease in the viability of H9c2 cells. Whereas, DOX-induced ROS accumulation and up-regulation of ERS related proteins were partially abolished by pretreatment with OP-D. Consequently, a DOX-induced ERS was mitigated by application of OP-D. Similarly, DOX-induced decrease in cell viability was partially attenuated by either inhibiting CHOP or pretreatment with N-acetylcysteine (NAC), an antioxidant. Moreover, cardiac ultrastructural abnormalities seen in mouse receiving DOX injections were obviously ameliorated by pretreatment of OP-D. Taken together, the present study proved that OP-D protects cardiomyocytes against DOX-induced injury, at least in part, through reducing ROS accumulation and alleviating ERS.
8.Research progress of mechanism of functional dyspepsia treated with acupuncture.
Kaiwei YAN ; Ling ZHAO ; Jie YANG ; Ying LAN ; Jing XU ; Chenchen WEI ; Fanrong LIANG
Chinese Acupuncture & Moxibustion 2015;35(9):973-976
Literature about functional dyspepsia treated with acupuncture in recent 5 years is retrieved in China National Knowledge Infrastructure (CNKI), Wanfang database and PubMed. The research achievements are arranged and summed up to explore the mechanism of acupuncture for functional dyspepsia. It is found that acupuncture can regulate the secretion of braingut petide, and cause the coordination response of limbic system-brain. Also, it adjusts serum molecule metabolin and the gene expression of the transduction pathway of adjustment signal for rats. It is believed that functional dyspepsia treated with acupuncture is through multiple ways, and adjusting the function of braingut axis is one of the important mechanisms.
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9.Recent research progress on swainsonine
Haiyun QUAN ; Zhenhui REN ; Hao LU ; Shuai WANG ; Ruixu XUE ; Jianguo WANG ; Chenchen WU ; Baoyu ZHAO
Chinese Journal of Veterinary Science 2017;37(8):1633-1640
Locoweeds are presently defined as those species of the genera Oxytropis and Astragalus (family Leguminosae) that specifically contain the key toxic constituent,swainsonine.After ingesting locoweeds,livestock can develop poisoning disease characterized by chronic dysfunction of the nervous system,which causes severe economic losses to the pastoral areas.In addition,swainsonine has attracted a great attention from toxicology and medicine fields,due to its dual role of toxicity and pharmacological activity.This review not only summarizes the latest research progress of toxicity and its poisoning mechanism,pharmacological activity,source,and biosynthesis pathway of swainsonine,but also speculates the possible regulatory enzymes involved in the synthesis pathway.Moreover,the future research on swainsonine is also looked ahead,which provide references for the prevention and treatment of locoism.
10.Analysis of the Quality of Prepared Slices of Herba Ephedrae
Kang CHEN ; Wenjin LIN ; Li LIN ; Chenchen ZHU ; Mudan ZHANG ; Chongyu ZHAO
Traditional Chinese Drug Research & Clinical Pharmacology 2000;0(05):-
Objective To compare the quality of prepared slices of Herba Ephedrae from different producing areas and to establish reference quality criterion of Herba Ephedrae.Methods Determined the content of Ephedrine and Pseudoephedrine in Herba Ephedra Prepared Herbal by HPLC.According to Chinese Pharmacopeia of 2000-year edition,extract,ash,water,impurity and foreign substance were detected; accelerated stability test was performed according to experience.Results The content of Ephedrine in Herba Ephedra is 0.995 %~1.589 %,honey-fried HerbaEphedra is 0.855 %~1.557 %;the content of Pseudoephedrine in Herba Ephedra is 0.560 %~2.087 %,honey-fried Herba Ephedra is 0.508 %~1.902 %; water-soluble extract was 8.83 %~18.30 %and 14.81%~27.45 %, alcohol-soluble extract was 7.74 %~18.83 %and 14.15 %~27.34 %, the total ash content was 6.49 %~10.29 %and 6.34 %~10.24 %and acid-insoluble ash was 0.19 %~0.42 %and 0.18 %~0.42 %in Herba Ephedrae and honey-prepared Herba Ephedrae respectively.The average water content was 8.10 %(s=0.3961)and 4.02 %(s=0.4674)and average content of impurity and foreign substance was 2.02 %(s=0.1954)and 2.01 %(s=0.2209)in Herba Ephedrae and honey-prepared Herba Ephedrae respectively.Conclusion This research will provide a reference criterion for the quality control of prepared slices of Herba Ephedrae.