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Objective:To investigate the effect of Chloriti Lapis on metal elements in brain tissue and plasma of epileptic rats kindled by pentylenetetrazol (PTZ), and to explore the possible material basis of Chloriti Lapis. Method:PTZ kindling method was used to establish epileptic rat model. Inductively coupled plasma mass spectrometry (ICP-MS) and inductively coupled plasma optical emission spectrometer (ICP-OES) were used to determine the contents of metal elements in brain tissue and plasma of the blank group, model group, carbamazepine group (0.1 g·kg<sup>-1</sup>) and Chloriti Lapis group (2 g·kg<sup>-1</sup>). The data were statistically analyzed by SPSS 18.0 software. Result:Compared with the blank group, the contents of Sr, Sb and Ba in brain tissue of rats in the model group were significantly increased (<italic>P</italic><0.05, <italic>P</italic><0.01), while the contents of Zn, Fe, Cu, K, Li, Co, Sn and Pb were significantly decreased (<italic>P</italic><0.05, <italic>P</italic><0.01). Compared with the model group, the contents of Zn, Fe, K, Li, Co, As and Pb in brain tissue of rats in the Chloriti Lapis group were obviously increased (<italic>P</italic><0.05, <italic>P</italic><0.01), while the contents of Sr and Sb were significantly decreased (<italic>P</italic><0.01). These results showed that Chloriti Lapis had positive effect on the regulation of the content of metal elements in rat brain tissue to normal level, the intervention effect was clear, and the overall effect was better than that of carbamazepine group. The determination of 21 metal elements in plasma showed that compared with the blank group, the levels of K, Sr and Cd in the model group were significantly increased (<italic>P</italic><0.05), and the contents of Li, Al, Ti and Cr were significantly decreased (<italic>P</italic><0.05). Compared with the model group, the contents of Ca, K, Li, Al and V in the Chloriti Lapis group were obviously increased (<italic>P</italic><0.05, <italic>P</italic><0.01), and the contents of Fe, Ti, Sr and Cd were significantly decreased (<italic>P</italic><0.05,<italic>P</italic><0.01). The correlation analysis of metal elements among the groups showed that there were 17 pairs of elements had positively correlation in the brain tissue of rats, 2 pairs of elements had significant negative correlation. In the plasma of rats, 8 pairs of elements had significant positive correlation and 6 pairs of elements had significant negative correlation. Conclusion:The metal element groups represented by Zn, Fe, K, Li, Co, As, Pb, Sr, Sb, Ca, Al, V, Ti and Cd may be the effective material basis for Chloriti Lapis to interfere PTZ-kindled epileptic model rats, which may be related to the influence of these metal element groups on the release of neurotransmitters and the electrical balance of neurons, the regulation of abnormal synchronous discharge induced by Na<sup>+</sup>, K<sup>+</sup>, Ca<sup>2+</sup> channel disorders and intervention of metabolism pathways in brain tissue related to epilepsy. It can make the excitatory and inhibitory activities restrain each other, and finally reach the normal physiological state of neurons and cells. The intervention effect of Chloriti Lapis group was better than that of carbamazepine group.
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Objective:To explore the possible mechanism of Chloriti Lapis in the treatment of epilepsy by the metabonomics of brain tissue in pentylenetetrazol (PTZ)-kindled epileptic rats treated with Chloriti Lapis. Method:The epileptic animal model in rats was established by PTZ kindling, and the rats were divided into the control group, model group, carbamazepine group and Chloriti Lapis group. The brain tissue samples were detected by ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC/Q-TOF-MS), and the experimental results were statistically analyzed by partial least squares-discriminant analysis (PLS-DA) and SPSS 18.0. Result:The metabolic fingerprints and metabolic profiles of the rat brain tissue were established, which showed that the metabolic profiles of each group had changed significantly and could be separated well among the groups. Moreover, the Chloriti Lapis group had a tendency to be closer to the control group than the carbamazepine group. Seven differential metabolites were screened, including phosphatidylserine (PS) (18∶0/18∶0), <italic>L</italic>-glutamic acid, docosahexaenoyl ethanolamide, arachidonic acid, glucosylsphingosine, cholestane-3,7,12,24,25-pentol and lysophosphatidylcholine (LysoPC) (P-18∶0). Except for docosahexaenoyl ethanolamide and LysoPC (P-18∶0), Chloriti Lapis had significant intervening and regulating effects on the other five differential metabolites. There were 12 possible metabolic pathways that affected the metabolic disorder of PTZ-kindled rats, and 3 important metabolic pathways (pathway impact>0.1), namely, <italic>D-</italic>glutamine and <italic>D-</italic>glutamate metabolism, alanine, aspartate and glutamate metabolism, and arachidonic acid metabolism, among which <italic>D-</italic>glutamine and <italic>D-</italic>glutamate metabolism was the most important metabolic pathways. Conclusion:From this point of view, Chloriti Lapis has a clear intervention effect on PTZ-kindled epileptic rats, which may be related to the intervention of the above differential metabolite contents and related metabolic pathways. It can reduce the toxic effect of excitatory neurotransmitters on neurons in brain tissue and inhibit the development of inflammation in brain tissue, so as to maintain the biological function of brain cells and slow down the occurrence of epilepsy.
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Objective:To study the effect of sedimentary type Limonitum on hemostatic indexes in blood and metal ions in serum of rats with hemorrhage. Method:The hemorrhagic rat models were established by warfarin sodium. The experimental animals were divided into control group,model group,powder group and water decoction group. On day 15 from drug administration, the contents of 6-keto prostaglandin F<sub>1</sub><italic><sub>α</sub></italic>(6-keto-PGF<sub>1</sub><italic><sub>α</sub></italic>),thromboxane B<sub>2</sub>(TXB<sub>2</sub>),arachidonic acid(AA),endothelin 1(ET-1),platelet activating factor(PAF),P-selectin(PS),and Ca<sup>2+</sup> in the whole blood of rats in each group were determined by enzyme-linked immunosorbent assay(ELISA). The contents of Na,Mg,K,Ca,Fe,Al,Li,Be,Ti,V,Cr,Mn,Co,Ni,Cu,Zn,As,Sr,Cd,Sn,Sb,Ba,and Pb in serum samples were determined by inductively coupled plasma optical emission spectrometer(ICP-OES) and inductively coupled plasma mass spectrometry(ICP-MS). Result:Compared with the model group,the content of 6-keto-PGF<sub>1</sub><italic><sub>α</sub></italic> was reduced in the powder group and water decoction group (<italic>P</italic><0.05),and the contents of TXB<sub>2</sub>,AA,ET-1,PAF,PS,Ca<sup>2+ </sup>were<sup> </sup>significantly increased(<italic>P</italic><0.01),with a positive and beneficial regulatory effect. In the powder group, 10 kinds of metal elements in serum of rats were significantly and positively regulated: Na,K,Ca,Fe,Li,Ti,V,Co,Cu,and Zn(<italic>P</italic><0.05,<italic>P</italic><0.01). In the water decoction group, 10 metal elements with significant positive regulation were as follows: Na,K,Ca,Fe,Li,V,Ni,Cu,Zn,and Sr(<italic>P</italic><0.05,<italic>P</italic><0.01). In addition,the content of Cr(<italic>P</italic><0.01) in the powder group and Cr(<italic>P</italic><0.01),Pb(<italic>P</italic><0.05) in the water decoction group were significantly reduced. Conclusion:The powder and water decoction of sedimentary type Limonitum had definite and positive intervention effect on warfarin hemorrhage model rats,which could play a coagulation role by enhancing the vasoconstriction ability,promoting the activation of platelets,and increasing the platelet aggregation rate and blood viscosity. The metal elements such as Na,K,Ca,Fe,Li,Ti,V,Co,Cu,Zn,Ni and Sr may be the material basis for sedimentary type Limonitum to exert hemostatic effect. According to the above indicators,the intervention effect of powder group and decoction group was basically the same.
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The effects of Chloriti Lapis on metal elements in plasma and lung tissue of acute exacerbation of chronic obstructive pulmonary disease( AECOPD) rats were studied. The rat AECOPD model with phlegm heat syndrome was established by smoking combined with Klebsiella pneumoniae infection. After the rats were treated by Chloriti Lapis,the contents of metal elements in plasma and lung tissue were determined by inductively coupled plasma-optical emission spectroscopy( ICP-OES) and inductively coupled plasma mass spectrometry( ICP-MS). The changes in the contents of metal elements were analyzed by SPSS 18. 0. Further,the correlations of differential metal elements( including Cu/Zn ratio) with differential metabolites in plasma,lung tissue and urine of AECOPD rats treated with Chloriti Lapis were analyzed. The results showed that Chloriti Lapis significantly up-regulated the contents of Fe,Al,Mn,Cu,Zn,Sn( P<0. 05),V,Co( P< 0. 01) and Cu/Zn ratio( P< 0. 05),and significantly down-regulated the contents of Ti( P< 0. 05)and Pb( P<0. 05) in the model rat plasma. It significantly increased the content of Be( P<0. 05) and decreased the contents of Mg,Ti and Al( P<0. 01) in model rat lung tissue. The element profiles of normal group,model group and Chloriti Lapis group can be well separated. Chloriti Lapis group and other groups were clustered into two categories. The taurine in plasma and phytosphingosine in lung tissue had the strongest correlations with differential metal elements. The Fe,Al,Mg,Be,Ti,V,Mn,Cu,Zn,Sn,and Co in Chloriti Lapis may directly or indirectly participate in the intervention of AECOPD rats. This group of metal elements may be the material basis of Chloriti Lapis acting on AECOPD rats,and reduce the Cu/Zn value in vivo. It was further confirmed that Chloriti Lapis could interfere with the metabolic pathways of taurine and hypotaurine in plasma and urine as well as the sphingolipid metabolism pathway in lung tissue of AECOPD rats. In addition,this study confirmed that long-term smoking can cause high-concentration Cd accumulation in the lung and damage the lung tissue.
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Animals , Rats , Lung , Medicine, Chinese Traditional , Minerals , Pulmonary Disease, Chronic Obstructive , Spectrum Analysis , Trace Elements/analysisABSTRACT
To study the effect of mineral Chloriti Lapis on pulmonary metabolites and metabolic pathways in lung tissues of rats with acute exacerbation of chronic obstructive pulmonary disease(AECOPD). The AECOPD rat model of phlegm heat syndrome was replicated by the method of smoking combined with Klebsiella pneumoniae infection. Except for using UPLC-Q-TOF-MS analysis, SPSS 18.0, SIMCA 13.0 and other software were also used for statistical analysis. Through literature search and online database comparison, the differential metabolites were identified, and the possible metabolic pathways were analyzed. After 15 days of administration, PLS-DA analysis was carried out on lung tissue samples of rats in each group. The results showed that the metabolic profiles of lung tissues of rats in each group could be well separated, which indicated that Chloriti Lapis and aminophylline had significant intervention effect on the lung metabolic profile of rats with AECOPD. Moreover, the metabolic profile of Chloriti Lapis group was closer to that of control group, and the intervention effect was better than that of aminophylline group. As a result, 15 potential differential metabolites were identified: phytosphingosine, sphinganine, tetradecanoylcarnitine, L-palmitoylcarnitine, elaidic carnitine, lysoPC[18∶2(9Z,12Z)], lysoPC(16∶0), lysoPC[18∶1(9Z)], lysoPC(18∶0), stearic acid, lysoPC(15∶0), arachidonic acid, docosapentaenoic acid, linoleic acid and palmitic acid. Among them, Chloriti Lapis could significantly improve the levels of 10 differential metabolites of phytosphingosine, tetradecanoylcarnitine, L-palmitoylcarnitine, elaidic carnitine, lysoPC[18∶2(9Z,12Z)], lysoPC(16∶0), lysoPC[18∶1(9Z)], stearic acid, lysoPC(15∶0), and palmitic acid(P<0.05). The intervention effect of Chloriti Lapis group was better than that of aminophylline group. Analysis of metabolic pathways showed that there were 8 possible metabolic pathways that could be affected, and three of the most important metabolic pathways(pathway impact>0.1) were involved: linoleic acid metabolism, arachidonic acid metabolism, and sphingolipid metabolism. Chloriti Lapis had obvious intervention effects on lung tissue-related metabolites and metabolic pathways in rats with AECOPD, and the effect was better than that of aminophyllinne.
Subject(s)
Animals , Rats , Lung , Medicine, Chinese Traditional , Metabolomics , Minerals , Pulmonary Disease, Chronic ObstructiveABSTRACT
Metallomics is a frontier interdisciplinary subject at its vigorous development stage. Its goal is to systematically study the content, distribution, chemical species, structural characteristics and functions of metal elements in biological system. It is also a comprehensive discipline to study the existing state and function of free or complex metal elements in life. Metallomics is an ideal tool to study the biological behavior of inorganic elements, which can be used to solve many problems in the research of mineral Chinese medicine(MCM). It provides a strong theoretical basis and technical support for the research of MCM. Its theory and methods provide re-ference and enlightenment for the in-depth study of MCM, and also provide new ideas and open up new ways for the research of MCM. The application of metallomics theory and methods in the research of MCM is of great significance to reveal the material basis and mec-hanism of MCM, promote the process of basic research on MCM, fully exploit and utilize medicinal mineral resources and carry forward the traditional MCM treasure in China. In this paper, we introduced the concept, academic development, research content and research methods of metallomics, and discussed the application prospects of metallomics in the analysis of inorganic element composition characteristics and quality control, material basis and mechanism of MCM, so as to provide reference for further researches on MCM.
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China , Drugs, Chinese Herbal , Medicine, Chinese Traditional , Minerals , Quality ControlABSTRACT
Objective: To provide the new quality control means for Alumen by investigating the elemental differences between calcined Alumen and its counterfeit processed products of ammonium alum, and establishing their characteristic chromatogram. Method: The contents of 22 inorganic elements both in calcined Alumen and processed products of ammonium alum were determined by means of inductively coupled plasma (ICP)-optical emission spectrometer-mass spectrometry (ICP-OES/ICP-MS),SPSS 16.0 was used for cluster analysis (CA) while SIMCA-P 13.0 with t-test and Rank-Sum test was used to identify the differential inorganic elements. In addition, the characteristic spectrum of the inorganic elements for calcined Alumen and counterfeit calcined alumen were established. Result: Calcined Alumen had highest contents of K and Al while counterfeit calcined Alumen has highest contents of Al and Fe;Cr,Sr,and Mn contents in calcined Alumen were relatively higher,while Mn,Ti,and Ga contents in processed products of ammonium alum were relatively higher. The content of K in calcined Alumen was about 205 times of that of counterfeit products. On the contrary,the average contents of Fe,Ti,Mn and Ga in counterfeit products of ammonium alum were much higher than those in calcined Alumen,33,46,38, 27 times, respectively. A total of 18 samples were clustered into two categories in CA:calcined Alumen and processed products of ammonium alum. 18 inorganic elements showed significant difference in contents(PConclusion: This method can be used for quality control of calcined Alumen.
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Objective: Ammonium alum is a common counterfeit of Alumen,and the processed product of ammonium alum is a common counterfeits of calcined Alumen. This paper aims to establish a method for identifying Alumen,calcined Alumen,ammonium alum and their processed products. Method: The samples were analyzed by scanning electron microscope (SEM) and X ray diffraction (XRD) in this paper. Result: Ammonium alum and Alumen showed obvious changes in morphology after processing. Both Alumen and ammonium alum showed obvious differences in morphology at×250 and×1 000 times microscope. Alumen presented irregular fragments,clear edge corners,smooth surface,scattered irregular small particles,occasional holes and longitudinal edges. Ammonium alum presented irregular clumps,blunt edges,not obvious edges and corners,uneven surface,scattered smaller and round-like particles. The difference in morphology was not obvious at×250 times microscope between Alumen and ammonium alum processed products. While at×1 000 times,the surface of calcined Alumen was uneven with coarse particles; the surface of counterfeit calcined Alumen was flat,and the coarse particle characteristics were not obvious. XRD can be used to rapidly and accurately identify the primary phase of Alumen,calcined Alumen,ammonium alum and ammonium alum processed products:KAl(SO4)2·12H2O,NH4Al(SO4)2·12H2O,KAl(SO4)2,and NH4Al(SO4)2 respectively, with 2θ angle characteristic value of 23,12,22 and 5 respectively for XRD peak. Conclusion: SEM and XRD techniques can be used for the identification of Alumen,calcined Alumen,ammonium alum and their counterfeit products.
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Objective: To evaluate the quality of Limonitum from different mineral genesis and select its high quality mineral resources by analyzing its mineral compositions and content. Method: The mineral compositions and content of different samples were analyzed by polarizing microscope and X-ray diffraction,and goethite content was taken as the evaluation index. SPSS 16.0 statistical software was used for hierarchical cluster analysis of the mineral compositions in different samples. Result: Limonite,quartz,illite and mica kaolinite were the main components of Limonitum from leaching disseminated type. Sedimentary type limonite was mainly composed of limonite,quartz and albite. Residual-alteration type limonite was mainly composed of limonite,quartz,diopside and carnallite. The content of goethite in limonite was highest (46.4%) in sedimentary type limonite from Jiangxi province,followed by that in leaching-disseminated type from Jiangsu province (17.9%) and leaching-disseminated type from Shandong province (0.5%). The content of sedimentary goethite was higher than 12.0%and the content of leaching-disseminated type goethite was between 0.5%and 18.0%, with significant difference; and the content of residual-alteration type was below 3.0%. Cluster analysis of mineral components in different samples showed that 6 samples of leaching-dissemination type were clustered into one group,3 samples of sedimentation type were clustered into one group,and 2 samples of residual-alteration type were clustered into one group. Conclusion: The quality of sedimentary type Limonitum in Jiangxi is the best. Generally,the quality of sedimentary type is better than that of leaching-disseminated type, and the quality of leaching disseminated type is better than that of residual-alteration type.
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Objective: To investigate the antidiarrheal effect and the gastrointestinal motility of two kinds of Limonitum with different mineral genesis, so as to select the optimal mineral genesis, and provide a theoretical basis for clinical use. Method: The selected research samples were leaching type and sedimentary type Limonitum with different mineral genesis, including powder, water decoction and dregs. In the experiment, castor oil (0.2 mL/10 g) was used to induce diarrhea, loperamide (4 mg·kg-1) was used for positive control, and then the antidiarrheal activity and the gastrointestinal motility were observed. Result: In the castor oil-induced diarrhea experiment, the diarrhea index of the water decoction (2.5,5,10 g·kg-1) and the dregs (2.5 g·kg-1) of leaching type Limonitum significantly reduced (P-1) of leaching type limonitum and the dregs (2.5,5,10 g·kg-1) of the leaching type Limonitum were significant different at the first defecation (P-1) and the water decoction (2.5 g·kg-1) of leaching type Limonitum, and the total feces weight of the mice were significantly decreased (P-1), the dregs (2.5,5,10 g·kg-1) of the sedimentary, the powder (2.5,5 g·kg-1), the water decoction (2.5,5,10 g·kg-1) and the dregs (10 g·kg-1) of leaching type Limonitum were significantly lower than those of the model group on the intestinal motility, with charcoal as a marker (P-1) of leaching type Limonitum was significantly higher than that of the model group on the content of small intestine contents (P-1) of sedimentary type limonitum, and the water decoction (2.5,5 g·kg-1) of leaching type Limonitum were significantly lower than those of the model group on gastric residual rate (P-1), the dregs (5 g·kg-1) of sedimentary type Limonitum were significantly reduced on the water content of large intestine compared with the model group (P-1 of Limonitum can inhibit the contraction of isolated ileum induced by acetylcholine in rabbits except 0.003 3 g·mL-1 water decoction of leaching type. Conclusion: Mineral medicine Limonitum has a certain therapeutic effect in castor oil-induced diarrhea. Leaching type limonitum has the best antidiarrheal effect.
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Mineral Chinese medicine is an indispensable part in traditional Chinese medicine, with a history of several thousand years in research and utilization of medicinal mineral resources. It is a summary of the medical experience accumulated in numerous trials,observations and practices during the survival process of people of all nationalities, showing distinctive characteristics. At present,the scope of mineral medicine treatment involves internal medicine,surgery,gynecology,pediatrics,facial features and so on. The mineral Chinese medicine has broad clinical application in many kinds of hemorrhagic diseases,with rich experience in medication and remarkable curative effects. However, there are few papers in modern research, mainly concentrated in the 1970s-1990s. In this paper, the pharmacology and clinical application of mineral Chinese medicine with hemostatic effects in the last 40 years were reviewed, aiming to provide references for its clinical rational use in bleeding diseases,further development and utilization of mineral Chinese medicine resources of traditional Chinese medicine,in-depth study of material basis and action mechanism,and further exploration of mineral Chinese medicine resources. According to the statistics of the major Chinese medicine works,27 kinds of mineral drugs have hemostatic effects. The researches on hemostatic pharmacology of mineral Chinese medicine mainly focus on the analysis of mineral medicine elements and the effect of related elements on hemostasis,as well as the pharmacological experimental study on hemostasis, but the research is not deep-going and comprehensive. The clinical application of mineral Chinese medicine for hemostasis mainly includes digestive tract hemorrhage,hemoptysis,epistaxis,gingival hemorrhage,cerebral hemorrhage,post-abortion hemorrhage,metrorrhagia,uterine bleeding,hematochezia and external hemostasis.
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Models were established in mice with warfarin sodium method, and their bleeding time and hemostasis time were measured by tail cutting method and slide method respectively. Rats were administered for 15 consecutive days to measure their recalcification time, plasma viscosity, platelet adhesion rate, platelet aggregation rate and other blood indexes. As compared with the blank group, the bleeding time was prolonged in model groupn(P<0.05). As compared with the model group, the results showed that the positive vitamin K, the leaching type water decoction and the sediment type decoction could significantly shorten the bleeding time (P<0.01); positive vitamin K significantly (P<0.01) shortened clotting time, and the leaching type water decoction, the sediment type water decoction and the sediment type powder could also shorten the clotting time (P<0.05). As compared with blank group, low dose, medium dose of leaching type water decoction, medium dose of powder, high dose of sediment type decoction and low dose of drug residues could reduce plasma viscosity (P<0.05), and high dose of leaching powder and low dose of water decoction could significantly reduce (P<0.01) plasma viscosity. As compared with blank group, Limonitum leaching type decoction high dose group could significantly reduce the platelet adhesion rate (P<0.05), while sediment type water decoction could significantly increase the platelet adhesion rate (P<0.05); the high dose of leaching type water decoction, high dose of drug residues, low dose of leaching type powder and low dose of drug residues could decrease the platelet aggregation rate (P<0.05), while high dose of leaching type water decoction and high dose of the powder could increase the platelet aggregation rate (P<0.05). Analysis of mineral compositions was conducted by polarized light microscopy and X-ray diffraction (XRD). The results of the both methods showed that Limonitum mineral compositions contained goethite, quartz, and kaolinite, and sedimentary type also contained illite and albite. Sediment type of Limonitum showed better hemostatic effect, which may be related to the high content of goethite and illite.