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ObjectiveTo observe the effects of Hedysari Radix polysaccharide on the apoptosis of gastric sinus smooth muscle cells and explore the underlying mechanism via the insulin-like growth factor-1 （IGF-1）/phosphatidylinositol 3-kinase （PI3K）/serine-threonine kinase （Akt） pathway in the rat model of diabetic gastroparesis （DGP）. MethodSixty-two Wistar male rats were randomized into a blank group （n=12） and a modelling group （n=50）. The rat model of DGP was established by small-dose multiple intraperitoneal injections of streptozotocin combined with an irregular high-fat and high-sugar diet for 4 weeks. The modeled rats were randomized into model group， mosapride citrate （1.35 mg·kg-1）， and high-， medium-， and low-dose （200， 100， and 50 mg·kg-1， respectively） Hedysari Radix polysaccharide groups. The rats were administrated with corresponding drugs by gavage， and those in the blank and model groups with equal volumes of pure water by gavage once a day for 8 consecutive weeks. The random blood glucose and body mass were measured every 2 weeks， and gastric emptying rate was calculated. Hematoxylin-eosin （HE） staining was used to observe the pathological changes of smooth muscle in gastric antrum， and terminal deoxynucleoitidyl transferase-mediated nick-end labeling （TUNEL） was employed to detect the apoptosis of smooth muscle cells in the gastric antrum. The expression of IGF-1， phosphorylated （p）-PI3K， and p-Akt in the smooth muscle of gastric sinus tissue was detected by immunohistochemistry. Western blot was employed to determine the protein levels of IGF-1， p-PI3K/PI3K， p-Akt/Akt， B-cell lymphoma-2 （Bcl-2）， and Bcl-2-associated X protein （Bax） in the smooth muscle of the gastric antrum. ResultCompared with the blank group， the model group showed elevated random blood glucose at all time points （P<0.01）， decreased body mass and gastric emptying rate （P<0.01）， increased apoptotic index of smooth muscle cells in the gastric antrum （P<0.01）， down-regulated protein levels of IGF-1， p-PI3K/PI3K， p-Akt/Akt， and Bcl-2， and up-regulated protein level of Bax （P<0.01）. Compared with the model group， the 8 weeks of drug administration lowered the random blood glucose， increased the body mass and gastric emptying rate （P<0.05， P<0.01）， decreased the apoptotic index of smooth muscle cells in the gastric antrum （P<0.05， P<0.01）， up-regulated the protein levels of IGF-1， p-PI3K/PI3K， p-Akt/Akt， and Bcl-2， and down-regulated the protein level of Bax （P<0.05， P<0.01）. Compared with the mosapride citrate group，the administration of low-dose Hedysari Radix polysaccharide for 6 and 8 weeks lowered the random blood glucose and decreased the body mass （P<0.05， P<0.01），low and medium-dose Hedysari Radix polysaccharide decreased the gastric emptying rate and the apoptotic index of smooth muscle cells in the astragaloside low-dose group decreased （P<0.05）. The protein levels of IGF-1，p-PI3K/PI3K，p-Akt/Akt and Bcl-2（low dose）were down-regulated and the protein level of Bax was up-regulated by low doses of Hedysari Radix polysaccharide （P<0.05， P<0.01）. Compared with high-dose Hedysari Radix polysaccharide， low-dose Hedysari Radix polysaccharide elevated random blood glucose and reduced body mass after 6 and 8 weeks of administration （P<0.05， P<0.01）， and the low and medium doses decreased the gastric emptying rate， increased the apoptotic index of smooth muscle cells in the gastric antrum （P<0.05， P<0.01）， down-regulated the protein levels of IGF-1， p-PI3K/PI3K， p-Akt/Akt， and Bcl-2， and up-regulated the protein level of Bax （P<0.05， P<0.01）. Compared with the medium-dose group，the low-dose group of Hedysari Radix polysaccharide had lower body mass，lower gastric emptying rate in rats，higher apoptotic index of smooth muscle cells in gastric sinus tissue after 6 and 8 weeks of administration （P<0.05， P<0.01）， and lower protein expression of IGF-1，p-PI3K/PI3K，p-Akt/Akt. ConclusionHedysari Radix polysaccharide protects the smooth muscle cells in gastric antrum against apoptotic injury and promotes gastric motility by activating the IGF-1/PI3K/Akt signaling pathway， as manifested by the up-regulated expression of IGF-1， p-PI3K， p-Akt， and Bcl-2 and down-regulated expression of Bax.
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Malignant tumors are one of the main causes of human death worldwide and pose a serious threat to human health. The current treatment methods are mainly the combination of chemotherapeutics， surgery， radiotherapy， or hormone therapy. The treatment process has limitations such as multidrug resistance， non-selective targeting of cancer cells， and drug toxicity. With the development and application of traditional Chinese medicine （TCM）， Chinese medicine has the characteristics of multi-angle and multi-mechanism coordination and slight toxic and side effects. It can effectively inhibit tumor proliferation， differentiation， and metastasis， and avoid drug resistance， serving as the focus of current tumor treatment research. Hedysari Radix， one of the genuine medicinal materials in Gansu province， is a tonic Chinese medicine with a wide range of pharmacological effects such as anti-inflammation， immune regulation， anti-oxidation， prevention and treatment of diabetic complications. In the majority of the ancient works on herbs of the past dynasties， Hedysari Radix was included under the item of Astragali Radix and used as Astragali Radix. Hedysari Radix is superior to Astragali Radix in enhancing immunity， scavenging free radicals， and resisting liver fibrosis. Studies have found that the effective components of Hedysari Radix have a prominent anti-tumor effect and a significant inhibitory effect on various malignant tumors such as liver cancer， bladder cancer， gastric cancer， breast cancer， and colorectal cancer. They can also combine with clinical anti-cancer drugs to reduce the toxic and side effects of chemotherapy drugs and improve the tolerance of patients during chemotherapy. On the basis of current research， this study summarized the mechanism of Hedysari Radix active components in inducing cell apoptosis， blocking cell cycle， inhibiting tumor cell proliferation， migration， and invasion， regulating micro mRNA （miRNA）， inducing cell autophagy， enhancing immune regulation， as well as reducing toxicity and enhancing efficiency and sensitization with clinical chemotherapeutics， and systematically explained the anti-tumor mechanism of Hedysari Radix active components， aiming to provide a basic reference for the further exploration of the anti-tumor mechanism of Hedysari Radix and the further development and utilization of its effective components.
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ObjectiveTo observe the effect of Hedysari Radix polysaccharide （HRP） on the Janus kinase 2 （JAK2）/signal transducer and activator of transcription protein 3 （STAT3） signaling pathway in diabetic nephropathy db/db mice. MethodFifty db/db mice were randomly divided into model group， irbesartan group （irbesartan suspension， 22.75 mg·kg-1）， and high-， medium-， and low-dose HRP groups （HRP suspension， 200， 100， 50 mg·kg-1） according to the body weight， with 10 mice in each group. Another 10 C57BL/6 mice were assigned to the normal group. The mice were treated with corresponding drugs by gavage， while those in the normal group and the model group received distilled water at 5 mL·kg-1. The mice in the six groups were administered once a day by gavage for 12 consecutive weeks. The uric acid （UA）， triglycerides （TG）， and total cholesterol （TC） were detected. Periodic acid-Schiff （PAS） staining and Masson staining were used to observe the pathological changes in kidney tissues. Western blot and Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） were used to detect the protein and mRNA expression levels of JAK2， STAT3， suppressor of cytokine signaling 3 （SOCS3）， and tumor necrosis factor-α （TNF-α） in the kidney. ResultAfter 12 weeks of treatment， compared with the normal group， the model group showed significant pathological ultrastructural changes in kidney tissues and increased UA， TG， and TC levels （P<0.01）. Compared with the model group， the high- and medium-dose HRP groups and the irbesartan group showed improvement in pathological ultrastructure of kidney tissues and reduced UA， TG， and TC levels （P<0.05， P<0.01）. Compared with the normal group， the model group showed a decrease in SOCS3 protein and mRNA expression levels and an increase in JAK2， STAT3， and TNF-α protein and mRNA expression levels （P<0.01）. Compared with the model group， the high- and medium-dose HRP groups and the irbesartan group showed an increase in SOCS3 protein and mRNA expression levels and a decrease in JAK2， STAT3， and TNF-α protein and mRNA expression levels （P<0.05， P<0.01）. ConclusionHRP can alleviate renal damage in diabetic nephropathy to a certain extent， and its mechanism may be related to the inhibition of the activation of the JAK2/STAT3 signaling pathway.
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This Radix study cum aims Melle to explore(HRPM)the on efficacy spleen differences deficiency between syndrome.modeling Astragali A Radix of Praeparata110cum rats Melle were(ARPM)randomized fatigue),and into rats Hedysari a Praeparata(n qi total irregular HRPM male diet,SD diarrhea,control were(n Yiqi=10)the=100).Pill group fied and model a modeling group,group Buzhong After(BYP)(through ARPM and the HRPM-H),classimedium-dose into(ARPM-M raised group,and high-dose(ARPM-H each and Rats BYP and under HRPM-M),normal and low-dose and(ARPM-L in and group HRPM-L)were groups,continuously10rats induced.were in group.the in group the were18.9,control given group were g·kg~(-1)conditions while those the the model Rats respectively in18.912.6,BYP kg~(-1)water extract,decoction those in ARPM/HRPM-H,the-M,dosage lasted and of-L groups treated the with control and model6.3group g·rewere motilin determined m L·kg~(-1)·day~(-1).days.of dose Spleen ARPM/HRPM of in water.morning,The at the10Rats spleen in index group thymus and index ceived equal calculated.(MTL),distilled tissue administration to15observe Then the and Routine of each group D-xylose,were was(IL-2),the subjected HE stainingγ(IFN-γ),lower to the pathological changes.(IgA),blood gastric indexes,mucosa index,interleukin-2group.interferon group immunoglobulin of A and spleen pepsin index,of in Ig A,IL-2spleen IFN-γ,control each MTL,levels Rats pepsin the in model(P<0.01),had higher levels routine(P<0.01),blood and indexes,more thymus lesions D-xylose,the and in index,level decreased HRPM-L of IL-2severe compared spleen with than the those model in group.thymus group.that(P<0.05group,P<0.01)index administration thymus groups Ig A or spleen as that and in spleen routine Except index,spleen the Ig A,index,group and were in in ARPM-M model group,group,index,indexes,P<0.01)and thymus MTL index,those in ARPM-L insignificantly Ig A,different pepsin from other those in the the blood index,compared IFN-γ,group,(P<0.05The D-xylose,model MTL,spleen and lesions high-dose in each administration administration groups group increased relieved.blood or comparison as of with HRPM in as the folARPM and the effect in and were white and result than ARPM and is of lows:(P stronger<0.05),of medium-dose high-dose HRPM HRPM on IL-2cell high-dose of(WBC)and count medium-dose the HRPM and corresponding doses than IFN-γmore ARPM the obvious effect(P<0.05of on evident(P<0.05of impact P<0.01),on low-dose between the on corresponD-xylose P<0.01),doses ding MTL doses than Meanwhile,in of or more high-dose,and medium-dose,difference HRPM the and indexes.corresponding there of ARPM in or IL-2no levels in the HRPM-L effect and two groups,on but conclusion,other the both functions IFN-γwas group no was difference more the than recovery that of the and ARPM-H between(IL-2,P<0.01;ARPM-L recovery HRPM the IFN-γ,P<0.05).HRPM-H and obvious therapeutic in rats group qi In ARPM dose have are certain equivalent,effects on with spleen function deficiency.the Specifically,is the better difference immunomodulatory of two at g·low kg~(-1).and but the promote immunomodulatory the of former rats significantly ARPM.than that between of the later two at in the dose>18.9HRPM promotion can of better digestion digestion absorption and may absorption due of than The immunoregulation and be to the difference in clinical medication.
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Animals , Rats , Astragalus Plant , Drugs, Chinese Herbal , Plant Roots , SpleenABSTRACT
Objective: To investigate the effects of five kinds of flavonoids (calycosin, formononetin, ononin, isoliquiritigenin, and medicarpin) from Hedysari Radix on promoting osteogenic differentiation of rat bone marrow stromal cells (rBMSCs) and rat calvarial osteoblasts (ROBs). Methods: rBMSCs were isolated according to plastic adherence. ROBs were isolated by enzyme digestion method. The proliferation of rBMSCs and ROBs were detected by MTT assay. ALP activity and calcium content of rBMSCs and ROBs cells were detected by alkaline phosphatase kit and calcium kit. Mineralized nodule formation was detected by alizarin red staining. Results: The five components could promote proliferation, increase ALP activity, increase calcium content, and increase the area and number of calcified nodules of rBMSCs and ROBs (P < 0.05). Among them, calycosin had the best effect on promoting the osteogenic differentiation of rBMSCs, and medicarpin promoted the osteogenic differentiation of ROBs with the best effect, followed by calycosin. Conclusion: Five flavonoids promoted the improvement of osteogenic function, while calycosin has better osteogenic activity on rBMSCs and ROBs and can be used as an excellent osteoinductive factor.
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Objective: To compare the effects of Hedysari Radix and Hedysari Radix Praeparata Cum Melle in Buzhong Yiqi and its chemical composition, and preliminarily reveal the mechanism of Hedysari Radix processed with honey. Methods: A rat model of spleen qideficiency was established. The rats were treated with different doses of water extracts of Hedysari Radix and Hedysari Radix Praeparata Cum Melle and the content of serum D-xylose, GAS, IL-2, TNF-α were used as indicators to study the differences in the efficacy of Hedysari Radix and Hedysari Radix Praeparata Cum Melle in Buzhong Yiqi. Based on HPLC techniques, the different components in methanol extract of Hedysari Radix and Hedysari Radix Praeparata Cum Melle were analyzed. Results: Compared with the blank group, the serum xylose, gastrin content in the model group were significantly decreased. Compared with the model group, except for the low dose group of Hedysari Radix, the serum xylose, and gastrin content of the rats in each administration group were significantly increased (P < 0.01), and the results in the middle dose group of Hedysari Radix were significantly higher than those in the middle dose group of Hedysari Radix Praeparata Cum Melle (P < 0.05, P < 0.01). Compared with the blank group, the serum IL-2 and TNF-α levels in the model group were significantly increased (P < 0.01). Compared with the model group, the serum IL-2 and TNF-α levels in the rats in each drug group were significantly decreased (P < 0.05, P < 0.01); Compared with the medium dose group of Hedysari Radix, the serum IL-2 and TNF-α levels in the medium dose group of honey-processing Hedysari Radix were significantly decreased (P < 0.05, P < 0.01). There were differences in the chromatographic peaks in the fingerprints of Hedysari Radix and Hedysari Radix Praeparata Cum Melle. Conclusion: Both Hedysari Radix and Hedysari Radix Praeparata Cum Melle can significantly intervene and improve the syndrome of spleen qi deficiency in rats and the pharmacodynamics effect of Hedysari Radix Praeparata Cum Melle is better than that of Hedysari Radix. There are differences in the components of Hedysari Radix and Hedysari Radix Praeparata Cum Melle and these differences may be the active substances that cause differences in the efficacy.
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Objective: To study the effect of small molecule compounds of Hedysari Radix in ntagonizing tumor necrosis factor receptor type 1 (TNFR1) based on molecular docking. Method: The structure of small molecular compound of Hedysari Radix was downloaded from the chemical composition compound library of traditional Chinese medicine, and then optimized to obtain the composition compound library of Hedysari Radix. The three-dimensional structure of the inflammatory target TNFR1 (PDB ID:1TNR) was identified. After hydrotreating and anhydrating, the binding pocket residues were identified according to the literature. According to the defined target structure and binding pocket, the flexible molecular docking was conducted between the composition compound library and the target, and the score (Glide Score) was obtained. Based on the results of molecular docking, the first nine small molecular compounds of Glide Score were selected as candidate components. On this basis, the drug-likeness was analyzed, which involved small molecular compounds that meet the number of hydrogen-bonded receptors, the number of hydrogen-bonded donors, the formula weight, the number of rotatable key and the numerical range of lipo-hydro partition coefficient. Finally, the binding mode was analyzed according to pharmacokinetic parameters and complex structure of composition-target docking. Result: The residue set in the TNFR1 drug-binding pocket were identified as glutamic acid109 (Glu109), lysine 35(Lys35), alanine62 (Ala62), serine 74 (Ser74), lysine75 (Lys75), cysteine76 (Cys76), argnine 77(Arg77), glutamine82 (Gln82), threonine89 (Thr89), asparticacid91 (Asp91), argnine92 (Arg92), aspartic acid93 (Asp93), threonine 94(Thr94), valine95 (Val95), cysteine 96(Cys96), argnine104 (Arg104), tyrosine106 (Tyr106), asparagine110 (Asn110), leucine111 (Leu111), phenylalanine112(Phe112), glutamic acid 131(Glu131) and lysine132 (Lys132). Totally 43 small molecular compounds of Hedysari Radix were obtained. Five small molecular compounds, namely hedysari radix, quercetin, isoliquiritin, naringenin, calycosin and liquiritigenin, were screened by comprehensive factors, like docking scoring. Conclusion: Quercetin, isoliquiritin, naringenin, calycosin and liquiritigenin are the effective anti-inflammatory substances of Hedysari Radix, with a great possibility of becoming TNFR1 antagonists.
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Objective Assessment of genetic diversity and genetic structure of germplasm of wild Hedysari Radix. Methods Fifteen germplasms of wild Hedysari Radix. were collected from Wudu pupolations and Tanchang pupolations. ISSR (Intersimple sequence repeat) markers were used. ISSR data were analyzed with the program POPGEN 1.32. The UPGMA tree and PCoA analysis was constructed using Ntsys software. The AMOVA analysis used Arlequin 31 software and Population structure analysis used the structure 2.3.4 software. Results The results of the ISSR-PCR showed that 126 (72.83%) of the 173 ISSR locitested were polymorphic by the nine primers. The change of percentage of polymorphic loci (PPL) of wild germplasm populations were 49.71%-61.85%. The average of PPL was 55.78%. Two populations of species level of observed number of alleles (Na), effective number of alleles (Ne), Nei's gene diversity (H), and Shannon's information index was 1.728 3, 1.364 6, 0.224 2, and 0.334 5, respectively. Total gene diversity (Ht), gene diversity with provenances (Hs), the coefficient of gene differentiation (Gst), and estimate of gene flow from Gst (Nm) of cultivated and wild population were 0.223 0, 0.192 1, 0.138 9, and 3.099 4, respectively. The among-population component accounted for 16.36% of the total variation, while the within-population component accounted for 83.64%, and genetic variation occured mainly within the population. UPGMA analysis showed that 15 samples were clustered into two branches including Ⅰ and Ⅱ. PCoA and population structure analysis confirmed the partitioning results of the UPGMA clustering. Mantel correlation test showed that there existed middle-level correlations between the genetic differentiation and geographical distance. Conclusion Our studies showed that the genetic diversity of Hedysari Radix populations was at a higher level, the characteristics of genetic structure included genetic differentiation that occurs mainly within populations and perennials, which provides theoretical basis for protecting and utilizing germplasm of Hedysari Radix resources.
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Objective To study the FT-IR fingerprint characteristics of Hedysari Radix from 8 producing counties in Gansu Province; To provide references for identification and application of Hedysari Radix in different producing counties. Methods FT-IR fingerprints of 110 batches of Hedysari Radix from 8 producing counties in Gansu Province were collected in the wave number range of 4000–400 cm-1. The common pattern of the fingerprints were analyzed, and the similarity analysis were used to analyze the FT-IR fingerprints of Hedysari Radix from 8 producing counties. The FT-IR fingerprint characteristics of Hedysari Radix from 8 producing counties in Gansu Province were compared. Results The rank of average similarity of FT-IR fingerprints of Hedysari Radix from 8 producing counties was Tanchang County > Li County > Xihe County > Wudu District > Zhang County > Min County > Longxi County >Weiyuan County, and Hedysari Radix from Longxi County and Weiyuan County were very different from other producing counties. The FT-IR fingerprints of Hedysari Radix from Longnan City (Tanchang County, Li County, Xihe County and Wudu District) were similar, and the average similarity was relatively high; while that from Dingxi City (Zhang County, Min County, Longxi County and Weiyuan County) were similar, and the average similarity was relatively low. Hedysari Radix from every producing county had a significant and unique FT-IR fingerprint characteristic. Conclusion The identification and application of Hedysari Radix from 8 producing counties in Gansu Province can be realized according to FT-IR fingerprint characteristics.
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Objective To establish a method for content determination of total flavones and polysaccharides in Hedysari Radix in different producing areas in Gansu Province.Methods The contents of total flavones and polysaccharides in Hedysari Radix in different producing areas in Gansu Province were determined by ultraviolet spectroscopy with calycosin-7-glucoside and glucose as reference substance, and the wavelength was set at 260 nm and 484 nm.Results The contents were from 2.82 mg/g to 6.79 mg/g for total flavones and from 106.14 mg/g to 746.40 mg/g for total polysaccharides in Hedysari Radix in different producing areas in Gansu Province. The recoveries of total flavones and total polysaccharides were 97.96％ and 102.90％, respectively.Conclusion There was difference in contents of total flavones and polysaccharides of Hedysari Radix in different producing areas in Gansu Province, and the method of using ultraviolet spectroscopy is simple, reproducible, accurate and reliable, which can be preferably used as the method for content determination of total flavones and polysaccharides in Hedysari Radix.
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Objective To compare the effects of different methods of honey sunburn on the contents of calycosin and formononetin in Hedysari Radix; To provide the basis for the establishment of the optimum processing technology. Methods By frying (traditional method), baking, and microwave methods put Hedysari Radix under honey sunburn. Agilent HC-C18 column (4.6 mm × 250 mm, 5 μm) was used; the mobile phase was acetonitrile-0.01% phosphoric acid solution, gradient elution (0–12 min, 30%–33% acetonitrile; 12–13 min, 31%–40% acetonitrile; 13–25 min, 40% acetonitrile) with velocity of 1.0 mL/min; detection wavelength was 248 nm;the column temperature was 30 ℃; sample volume was 10 μL. Results There was statistical significance in the contents of calycosin and formononetin of different methods of honey sunburn for Hedysari Radix. Among them, the contents of calycosin and formononetin in Hedysari Radix processed by honey roast were the highest, 7.9116 and 49.6996 μg/g, respectively; the contents of calycosin and formononetin in Hedysari Radix processed by traditional method were the lowest, 4.7767 and 37.2910 μg/g, respectively; the contents of calycosin and formononetin in raw Hedysari Radix and Hedysari Radix by honey microwave method were the same, 5.0802, 42.7989 μg/g, and 3.9839, 42.3145 μg/g, respectively. Conclusion Different honey sunburn methods for the contents of calycosin and formononetin in Hedysari Radix have certain effects, and honey roast method is the optimum method.
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Objective To establish a method for simultaneous determination for the contents of four flavones (ononin, calycosin, genistein and formononetin) of Hedysari Radix in Gansu Province with quantitative analysis of multi-components by single-marker (QAMS); To prove its feasibility and accuracy. Methods Calycosin was taken as internal standard substance. Relative correction factors (RCF) of ononin, genistein and formononetin to calycosin were established. The contents of ononin, calycosin, genistein and formononetin were determined to realize QAMS. Results RCF was with good repeatability. The results of QAMS were consistent with the results of the external standard method. Conclusion The method that determines the contents of ononin, genistein and formononetin with calycosin as internal standard substance, can be used for quantitative analysis of Hedysari Radix.
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Leguminous related SSR primers were collected, core primers used for Astragali Radix and Hedysari Radix identification were screened and validated by using molecular marker techniques. 6 core primers were selected from 101 pairs of primers, the molecular weight of PCR products was 100-500 bp, which formed 7-12 electrophoresis bands with 55 amplified loci. The percentage of polymorphic loci was 100%, and the average polymorphism information content was 0.371. According to the results of cluster analysis, obtained core primer could completely distinguish 62 mixture samples of Astragali Radix and Hedysari Radix in similarity coefficient of 0.46. Core primers and the corresponding characteristics from gel electrophoresis were tagged. The results provide identification basis for Astragali Radix and Hedysari Radix.
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This paper was aimed to compare the effect of Buzhong Yiqi decoction containing Hedysari Radix or Astragali Radix on anti-immunosenescence effects in spleen lymphocytes of senescence accelerated mouse 8 (SAMP8). The effect of the serums on the proliferation of spleen T lymphocytes in SAMP8 mice induced by ConA was tested by MTT. The effect of the serums on the T lymphocytes subsets of SAMP8 mice was measured by flow cytometry. ELISA was used to detect the level of IL-2 and IFN-γ in the culture supernatants of spleen lymphocytes. The effect of the serums on the expression of CD28 mRNA in spleen T lymphocytes was detected by fluorescent quantitative PCR. Western blot was used to detect the expression of CD28 protein in spleen T lymphocytes of SAMP8 mice. Both the serums of Buzhong Yiqi decoctions containing Hedysari Radix or Astragali Radix improved the proliferation of T lymphocytes in SAMP8 mice. Both the serums had no obvious effect on the differentiation of spleen T lymphocytes'subsets in SAMP8 mice. Both the serums increased the content of IL-2 and INF-γ in the culture supernatants of spleen lymphocytes. And for the content of IL-2, the serum of Buzhong Yiqi decoction with Hedysari Radix was better（P<0.05）. Both the serums improved the expression of CD28 mRNA in spleen T lymphocytes of SAMP8 mice. And the effect of Hedysari Radix group was better than that of Astragalus Radix group（P<0.05）. Both the serums improved the expression of CD28 protein in spleen T lymphocytes of SAMP8 mice. The role of the serums containing Buzhong Yiqi decoction with Astragalus Radix and the decoction with Hedysari Radix in anti-immunosenescence was through the effect of the CD28. And the effect of Hedysari Radix group was better than that of Astragalus Radix group on improved the expression of CD28 mRNA in T lymphocytes of SAMP8 mice. Astragalus Radix and Hedysari Radix could swap in the aspect of anti-immunosenescence.
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Objective To discuss the correlativity of the characteristics of trace elements and the quality in Hedysari Radix in different growth ages by taking the Hedysari Radix with different growth ages in Gansu as the research objects. Methods Hedysari Radix in different growth ages was collected. The contents of 12 trace elements in Hedysari Radix were detected by using flame atomic absorptions pectrophotometer.The multiple comparisons, correlation analysis, discriminant analysis and factor analysis were performed by SPSS21.0. Results The contents of Ni, Mg, and Mn were significant in Hedysari Radix with different growth ages. The growth ages had significantly negative correlation with Zn contents and had significantly positive correlation with Li contents. The contents of Cu, Fe, Ca, K, Zn, Na, and Mn and growth ages existed Fisher linear. The quality of Hedysari Radix with different growth ages had difference;the quality of Hedysari Radix with 2-year-old growth ages also had difference; the quality of 3 and 4-year-old Hedysari Radix was better than 6 and 8-year-old. Conclusion There are closely relationships between growth and the contents of certain trace elements. The quality of Hedysari Radix has some relationships with the contents of trace elements.
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Objective To analyze the characteristic changes of trace elements from Hedysari Radix in different months;To provide references for the choice of the best harvest periods. Methods Trace elements of Hedysari Radix from different months (June, July, August, September, October and November) were determined by atomic absorption spectroscopy and analyzed by single factors analysis of variance, correlation analysis, cluster analysis and factors analysis. Results There was obvious significance between the contents of Cu and Co (P<0.05), and there was obvious significance among the contents of Fe, Ca, Mn, Zn, Mg, Cr, Na, Li, Ni and K (P<0.01). The results of correlation analysis showed that months had positive correlation with Cu, had a negative correlation with Fe, Na, Mn, Cr and Li contents, and had significantly negative correlation with Ca, Mg contents. Mn, Cr, Ni, K and Na were the characteristics of trace elements from Hedysari Radix in different months. The different months samples could be divided into two categories of June, July, August, September samples (Ⅰ) and October, November samples (Ⅱ). Conclusion There is close relationship between the contents of trace elements from Hedysari Radix and months. The information and metabolic patterns of trace elements from Hedysari Radix in June, July, August, September (immature) samples are close or similar;the information of trace elements in October, November (nearly mature or maturing) samples are close or similar. Study on harvest times of Hedysari Radix by trace element as an indicator is with certain rationality.
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ObjectiveTo observe the effects of ultrafiltration extract from Angelica Sinensis Radix and Hedysari Radix on PC12 cell apoptosis induced by H2O2; To discuss its mechanism of action.Methods H2O2 was used in the incubation of PC12 cells to establish the oxidative damage nerve cell apoptosis model. The experiment was divided into normal control group, model group, and three different dosages (0.375, 0.75, 1.5 g/L) of ultra-filtration extracted from Angelica Sinensis Radix and Hedysari Radix after modeling for interference. Rate of cell apoptosis was detected by flow cytometry; mitochondrial membrane potential was measured by laser scanning confocal microscopy; the protein expressions of cleaved Caspase-3, Bax and Bcl-2 were detected by Western blot.Results Compared with the model group, rates of cell apoptosis in the different dosage groups decreased significantly; membrane potential of mitochondria increased; the protein expressions of cleaved Caspase-3 and Bax decreased; the expression of Bcl-2 increased; the ratio of Bcl-2 and Bax increased (P<0.05).Conclusion The ultra-filtration extracted from Angelica Sinensis Radix and Hedysari Radix can inhibit PC12 cell apoptosis induced by H2O2.
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Objective To evaluate the quality of Hedysari Radix from different producing areas in Gansu Province by setting Hedysari Radix from different producing areas in Gansu Province as study subjects and trace elements in Hedysari Radix as evaluation indexes; To provide the references for establishment of the quality standards of Hedysari Radix.Methods The content of Fe, Cu, Ca, Mn, Zn, Mg, Cr, Co, Na, Li, Ni and K in Hedysari Radix were detected by using flame atomic absorbption method. Data were analyzed by the factor analysis conducted by SPSS21.0 software. The common factors were extracted by principal component and analytic function for quality comprehensive ranking was established.ResultsThree common factors were extracted, namely F1, F2, and F3. The quality function was Y=0.657F1+0.119F2+0.089F3. Cr, Li, Co, Cu, Zn, Fe and Mg were the characteristic trace elements of Hedysari Radix that could affect the quality of Hedysari Radix. When the quality of Hedysari Radix was evaluated by the contents of trace elements, the quality of wild Hedysari Radix were better than cultivated Hedysari Radix and the quality rank was consistent with the idea that Longnan is the main producing area of Hedysari Radix. The quality of samples from Dingxi area was better.Conclusion The quality evaluation of Hedysari Radix based on trace elements shows the differences in Hedysari Radix from different producing areas in Gansu Province and the results are relatively reasonable.
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Objective: To study the correlation of HPLC fingerprint of Hedysari Radix and efficacy of improving immunity of normal mice, and provide the basis for finding bioactive components and selecting the index composition of quality control. Methods: The mice delayed type hypersensitivity (DTH) model induced by dinitoflruorobenzene (DNFB) was used to study the effect and then to perform ig administration with different extracts of Hedysari Radix for selecting the best part. To study the correlation between fingerprint and efficacy and find out the effective substances foundation of improving immunity, the peak area of each common peak from HPLC fingerprint was associated with the date of improving immunity in the method of partial least square regression (PLS). Results: In ear swelling experiment, water extract of Hedysari Radix was the most effective part which could improve the immunity function. PLS was used to evaluate the correlation of effect of improving immunity and components of the water extract including adenosine, ononin, genistein, formononeti, and medicarpin, the index of compounds to characterize the quality of Hedysari Radix, showed the negative correlation with the effectiveness. Conclusion: A new method is established to evaluate the activity of enhancing immunity of Hedysari Radix. It is of practical significance as an effective approach for controlling quality and exploring the material basis for efficacy of traditional Chinese medicine compounds.
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Objective: To investigate the protective effect of Hedysari Radix polysaccharide (HRP) on liver fibrosis in mice and the correlation between the hepatic fibrosis and bone loss. Methods: The mice were sc injected with 40% carbon tetrachloride (CCl4) dissolved in olive oil solution (0.1 mL/kg, once every 5 days ), while given high-, mid-, and low-dose (20, 10, and 5 g crude drug/kg) HRP and colchicines (0.6 mg/kg) daily. Continuous 35 d later, the activity of alanine transarninase (ALT) and aspartate transferase (AST), contents of total protein (TP) and albumin (ALB) were investigated by related reagent kit, and then, calculated the albumin/globulin (A/G) ratio, liver and thymus indexes and liver histological of tissue pathology were determined. The serum level of acid phosphatase resistant to acid phosphatase (TRACP), alkaline phosphatase (ALP), Ca, P, and the bone level of hydroxyproline (Hyp) were investigated by related reagent kit to observe bone loss. Results: The model group compared with the Sham group, the levels of ALT, AST, and TP increase, showed obvious liver pathological damage, liver and thymus index increase, levels of TRACP, ALP, Ca, and P decrease, and Hyp increase. HRP could significantly improve the hepatic fibrosis and bone loss induced by CCl4 in mice. Conclusion: The liver fibrosis in mice could also cause bone loss and be the primary cause of bone loss in this experiment. The certain market value of this Chinese materia medica is developed for its significant effect and contact between hepatic fibrosis and bone loss is initially investigated, which lays the foundation to further discuss the mechanism of the above two.