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OBJECTIVE To optimize ethanol extraction process of Yihuang powder. METHODS An orthogonal experiment was designed by reflux extraction with ethanol volume fraction， liquid-to-material ratio， and extraction time as investigation factors. The parameters used were the contents of hesperidin， nobiletin， tangeretin， gallic acid， chebulagic acid， chebulinic acid， liquiritin， glycyrrhizin， eugenol， and the paste-forming rate. The analytic hierarchy process （AHP） was used to calculate the comprehensive score. The optimal ethanol extraction process parameters of Yihuang powder were determined by verifying the results predicted by orthogonal experiment and genetic algorithm （GA）-back propagation neural network （BP neural network）. RESULTS The optimal ethanol extraction process parameters， as optimized by orthogonal experiment， were as follows: ethanol volume fraction of 60%， liquid-solid ratio of 14∶1 （mL/g)， extraction time of 90 min， and extraction for 2 times. The comprehensive score obtained by verification was 79.19. Meanwhile， the optimal ethanol extraction process parameters， optimized by GA-BP neural network， were ethanol volume fraction of 65%， liquid-solid ratio of 14∶1 （mL/g )， extraction time of 60 min， and extraction for 2 times. The comprehensive score obtained by verification was 85.30， higher than the results obtained from orthogonal experiment. CONCLUSIONS The optimization method of orthogonal experiment combined with GA-BP neural network is superior to the traditional orthogonal experiment optimization method. The optimized ethanol extraction process of Yihuang powder is stable and reliable.
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This paper aimed to explore the anti-inflammatory effect of ethanol extract from Saposhnikoviae Radix in a lipopolysaccharide(LPS)-induced inflammation mouse model and its regulation of TLR4/NF-κB signaling pathway. The ethanol extract from Saposhnikoviae Radix was separated and purified on the macroporous adsorption resin and its main chemical components were identified by UPLC-QE/MS. The identification results showed that the top ten components of ethanol extract from Saposhnikoviae Radix were mainly chromones and coumarins. A mouse model of inflammation induced by intraperitoneal injection of LPS was used to investigate the anti-inflammatory effects of ethanol extract from Saposhnikoviae Radix after intragastric administration for seven successive days. Mice in all groups except for the control group were treated with intraperitoneal injection of LPS(0.015 g·kg~(-1)) one hour after the last administration, and twelve hours later, the blood was sampled and separated and the broncoalveolar lavage fluid(BALF) was collected. The levels of nitric oxide(NO), tumor necrosis factor-α(TNF-α), interleukin-6(IL-6), and interleukin-1β(IL-1β) in mouse serum and BALF were detected by ELISA. The harvested lung tissue was stained with hematoxylin-eosin(HE) for observing the pathological changes, followed by the detection of protein expression levels of related molecules in TLR4/NF-κB signaling pathway by Western blotting. The results showed that the ethanol extract from Saposhnikoviae Radix significantly ameliorated the pathological conditions in lung tissue of model mice, reversed the increase in NO, TNF-α, IL-6, and IL-1β levels of mouse serum and BALF, down-regulated the protein expression levels of Toll-like receptor 4(TLR4), myeloid differentiation factor(MyD88), and phosphorylated nuclear transcription factor κB-p65/nuclear transcription factor κB-p65(P-NF-κB p65/NF-κB p65), and up-regulated the NF-κB inhibitory protein α(IκBα). The ethanol extract from Saposhnikoviae Radix exhibited a good anti-inflammatory effect in the LPS-induced acute inflammation muse model, which might be related to the inhibition of the activation of TLR4/NF-κB inflammatory signaling pathway. Chromones and coumarins have been proved to be the active components for its anti-inflammatory effects.
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Animals , Mice , Anti-Inflammatory Agents , Ethanol , Inflammation/drug therapy , Lipopolysaccharides/toxicity , NF-kappa B/genetics , Plant ExtractsABSTRACT
BACKGROUND: Studies have found that Sambucus sibirica Nakai can promote osteoblast division, proliferation, and secretion of alkaline phosphatase to accelerate bone matrix calcification, promote new bone formation, and promote the absorption of necrotic bone. OBJECTIVE: To observe the effects of two extracts of Xinjiang Sambucus sibirica Nakai bark in promoting fracture healing in rats. METHODS: A total of 108 male Sprague-Dawley rats, 3 months old, were randomly divided into sham-operated group, model group, Zhong Hua Die Da Wan group (0.54 g/kg, negative control group), high-, middle- and low-dose ethanol extract groups (0.314, 0.157, 0.078 g/kg), high-, middle, and low-dose water extract groups (0.340, 0.170, 0.085 g/kg). Except for the sham-operated group, the middle segment of the femoral shaft was cut off to make a model of open femoral fracture. In the sham-operated group, only the femoral shaft was exposed and no osteotomy was performed. Intragastric administration began at 1 day after modeling and continued for 6 weeks (1 time/day). At 2, 4, 6 weeks after modeling, the effects of two extracts of Sambucus sibirica Nakai bark on serum-related biochemical indicators, X-ray imaging, biomechanical indicators and hematoxylin-eosin staining were observed. RESULTS AND CONCLUSION: (1) Serum biochemical indicators: The serum calcium level of the high-dose ethanol extract group in each phase was lower than that of the sham-operated group and the model group, indicating that a faster osteogenesis in the high-dose ethanol extract group, followed by middle-dose ethanol extract group, low-dose ethanol extract group, and high-dose water extract group. The serum phosphorus level of ethanol extract groups was higher than that of sham-operated group, model group, and Zhong Hua Die Da Wan group; the serum alkaline phosphatase level of high-dose ethanol extract group at each stage was significantly higher than the other groups, indicating that a stronger osteoblast activity in the high-dose ethanol extract group, followed by the high-dose water extract group. (2) X-ray film: Fracture healing was significant in the high-dose ethanol extract group, fair in the Zhong Hua Die Da Wan group, high-dose water extract, middle-dose water extract group, middle-dose ethanol extract group, and poor in the low-dose ethanol extract group and low-dose water extract group. (3) Three-point bending test: At 6 weeks, the maximum load and maximum deflection of the femurs of the two extract groups were higher than those in the model group. The maximum load and maximum deflection of the femurs increased significantly in the high-dose ethanol extract group followed by the high-dose water extract group, middle-dose water extract group and middle-dose ethanol extract group. (4) Hematoxylin-eosin staining: Fibrous epiphysis gradually formed in the fracture area, and the bone marrow cavity was communicated. Bone healing was better in the high-dose ethanol extract and high-dose water extract groups than in the other groups. To conclude, these two extracts of Xinjiang Sambucus sibirica Nakai bark are capable of improving fracture healing. Moreover, different extraction methods and dosages of Sambucus sibirica Nakai bark demonstrate various effects on fracture healing. Compared with water extracts, ethanol extracts are better to promote fracture healing, and high-dose extracts have better effects than middle- and low-dose extracts.
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Objective: To explore the therapeutic effect of cistanche deserticola ethanol extraction (CDE) in the rats with osteoporosis induced by ovariectomy, and to clarify its mechanism. Methods: A total of 72 female SD rats were randomly divided into control, model, estradiol (0. 3 g · kg-1), low, middle and high doses (0. 5, 1.0 and 2. 0 mg · kg-1) of CDE groups, and 12 rats in each group. The rat ovaries were removed by operation in the other groups to establish the osteoporosis models, while the same size of fat of the rats in control group was excised. 5 d later, the rats in estradiol group were hypodermicly injected with estradiol, and the rats in CDE groups were orally administrated with corresponding doses of CDE. The same volume of diluted water was given to the rats in control and model groups. After 20 weeks, the density of right femur of the rat was detected with the method of ratio of weight and size; the mechanics index of left femur of the rat was analyzed by bone granulometer; the levels of serum alkaline phosphatase (ALP) and osteocalcin were measured by ELISA; the levels of serum Ca2 and P were detected by automatic biochemical analyzer; the morphology of uterus tissue was observed by HE staining. Results: Compared with model group, the densities of right thighbone of the rats in estradiol group and low, middle and high doses of CDE groups were significantly increased (P0. 05). Compared with model group and low dose of CDE group, the maximum bending forces, the maximun strains, and the bone rigid coefficients in middle and high doses of CDE groups were remarkably increased (P0. 05). Compared with model group, the morphology of uterus tissue of the rats in low, middle and high doses of CDE groups was improved at different degrees. Conclusion: CDE has therapeutic effect on osteoporosis through increasing the levels of serum ALP, osteocalcin and Ca' of the rats.
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Objective:To explore the therapeutic effect of cistanche deserticola ethanol extraction (CDE) in the rats with osteoporosis induced by ovariectomy,and to clarify its mechanism.Methods:A total of 72 female SD rats were randomly divided into control,model,estradiol (0.3 g · kg-1),low,middle and high doses (0.5,1.0 and 2.0 mg · kg-1) of CDE groups,and 12 rats in each group.The rat ovaries were removed by operation in the other groups to establish the osteoporosis models,while the same size of fat of the rats in control group was excised.5 d later,the rats in estradiol group were hypodermicly injected with estradiol,and the rats in CDE groups were orally administrated with corresponding doses of CDE.The same volume of diluted water was given to the rats in control and model groups.After 20 weeks,the density of right femur of the rat was detected with the method of ratio of weight and size;the mechanics index of left femur of the rat was analyzed by bone granulometer;the levels of serum alkaline phosphatase (ALP) and osteocalcin were measured by ELISA;the levels of serum Ca2+ and P were detected by automatic biochemical analyzer;the morphology of uterus tissue was observed by HE staining.Results:Compared with model group,the densities of right thighbone of the rats in estradiol group and low,middle and high doses of CDE groups were significantly increased (P＜0.05 or P＜0.01);there were no differences between low,middle and high doses of CDE groups (P＞0.05).Compared with model group and low dose of CDE group,the maximum bending forces,the maximun strains,and the bone rigid coefficients in middle and high doses of CDE groups were remarkably increased (P＜ 0.05),while the levels of ALP were decreased (P＜ 0.05).Compared with model group,and low and middle doses of CDE groups,the level of osteocalcin in high dose of CDE group was significantly decreased (P＜0.05).Compared with model group,the levels of Ca2+ in serum of the rats in low,middle and high doses of CDE groups were significantly increased (P＜0.05);there were no significant differences between low,middle and high doses of CDE groups (P＞0.05).Compared with model group,the morphology of uterus tissue of the rats in low,middle and high doses of CDE groups was improved at different degrees.Conclusion:CDE has therapeutic effect on osteoporosis through increasing the levels of serum ALP,osteocalcin and Ca2+ of the rats.
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Objective To optimize the ethanol extraction technology ofLichong ShengsuiDecoction.Methods An L9(34) orthogonal test was used in the study. The extraction rates of calycosin glycoside, icariin, baohuosideⅠ, ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1, astragaloside, and ginsenoside Rd were set as indexes. The influence of ethanol concentration, extraction time, extraction temperature, and amount of ethanol on the yield of Lichong ShengsuiDecoction were detected by comprehensive scoring method.Results The optimal ethanol extraction technology forLichong ShengsuiDecoction was soaking for 2 h with ten times of 60% ethanol and then reflux extracting for two times; extraction time was 1.5 h each time at 80℃.ConclusionThe optimal extraction technology is efficient, stable and feasible, which can provides data for the further study ofLichong ShengsuiDecoction.
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The effective exploitation of natural products is of great significance. Herein the essential oil from Herba Artimisiae Sieversianae in growth period and flower/ fruit bearing period was extracted by ethanol extraction method. The optimal extraction condition was determined by orthogonal experiment, including extracting 3 times, soaking for 90 min, and the ratios of liquid to solid for Herba Artimisiae Sieversianae in growth period and flower/ fruit bearing period were 10 mL : 1 g and 8 mL : 1 g, respectively. The main components of the products were confirmed by FT-IR and GC-MS, which were cineole, camphor, d-borneol, caryophyllene, cadina-1, 4-diene, calamenene, ethyl palmitate, etc. in herba artimisiae sieversianae in growth period, and camphor, caryophyllene, borneol, neryl formate, etc. in Herba Artimisiae Sieversianae in flower/ fruit bearing period. The radical scavenging activity of the products was determined. The results showed that the IC50 values of the essential oil from Herba Artimisiae Sieversianae in growth period and flower/fruit bearing period for 0. 05 mmol/ L DPPH solution were 0. 40 mg / mL and 1. 66 mg / mL, respectively. The essential oil extracted by ethanol from Herba Artimisiae Sieversianae was confirmed to possess good antioxidant activity.
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To optimize the ethanol extraction process for Shenlian formula. On the basis of the pharmacodynamics index for different extraction process routes, the contents of salvianolic acid B, tanshinone ⅡA and berberine, as well as the extraction ratio in different experimental schemes were used as the ethanol extraction examining indexes, and multi-criterion synthesizing grading method was used for data analysis to optimize and verify the ethanol-extraction process conditions in orthogonal experiment. The optimum ethanol extraction process was as follows: adding 60% ethanol, 10 times amount, extracting for 2.0 h each time for a total of 2 times. This extraction process showed good stability and availability.
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Objective To optimize ethanol extraction technologyDingtongning Granule.Methods With yield of extract and content of tanshinoneⅡA as comprehensive evaluation indexes, the concentration of ethanol, the amount of ethanol, soaking time, extraction time and extraction times were selected as observing factors. Orthogonal design test L18(37) was used to optimize ethanol extraction technology ofDingtongning Granule.Results Optimum extraction conditions were as follows: soaking medicine for 0 hour; 2 times with 6 times of the amount of 80% ethanolre; flux extracted 3 hours each time.Conclusion This optimal extraction technology is stable and reliable, which can provide basis for industrial production ofDingtongning Granule.
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Objective: To optimize the extracting and processing conditions of Compound Jianshen Granules (CJG). Methods: UV analytical methods of total saponin and total polysaccharides were established, the optimal ethanol extraction process and water extraction process were confirmed according to single factor and orthogonal design test. The influence of different excipients and wetting agents were investigated, taking forming rate, solubility, and absorption rate as indexes, the materials added prescription varieties, ratio, and optimum molding process were screened. Results: The optimal ethanol extraction technology of CJG was: 85% ethanol, 10-fold, extracting for three times, 2 h once. The optimal water extraction technology was 14-fold, decocting 3 times, 30 min once. The optimized conditions of CJG were obtained with: accessories for the dextrin: lactose (4:1), main drug-accessories (1:1), with 85% ethanol as wetting agent which accounted for 15% of the main drug. Conclusion: The extraction process and forming process are feasible and stable, and the quality is controllable.
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Female rats were fed with ethanol extraction of Alismatis Rhizoma for 6 months to study its nephrotoxicity and molecular mechanism. HPLC was used to determine the components in ethanol extraction of Alismatis Rhizoma. An assessment of renal pathology was determined by HE staining. Meanwhile Western blot, immunohistochemical assay and q-PCR were used to assess the protein expression and mRNA level of Kim-1, clusterin, LCN2, osteopontin, ceruloplasmin and TIMP1 in rat kidney. Eight components were identified in ethanol extraction of Alismatis Rhizoma. Tubule-interstitial inflammation, renal tubular epithelial cell exfoliation and morphological changes of cell were observed in rat kidney. Comparing with control blank group, the protein expression of clusterin, Kim-1, LCN2, osteopontin and TIMP1 in rat kidney was significantly increased while the protein expression of ceruloplasmin was significantly decreased. The mRNA level of Kim-1, TIMP1, osteopontin, clusterin and LCN2 was significantly increased while the mRNA level of ceruloplasmin was significantly decreased. In this study, it was inferred that there is chronic toxicity in kidney by using high dosage of ethanol extraction of Alismatis Rhizoma for a long time. And the underlying molecular mechanism was related to regulate the protein expression of ceruloplasmin, clusterin, Kim-1, LCN2, osteopontin and TIMP1.
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Objective To optimize ethonal extraction technology for no sugar runfei granules.Methods Ethanol was used as a solvent to extract the ethanol dissolvable ingredients in 9 kinds of medicinal materials.The formula L9 (34 ) table was used to examine the effects of 4 factors and 3 levels.The weighted sum of baicalin content and the dried extract quantity was used as quantitative index.Results The maximize optimize condition for extraction of ethanol dissolvable ingredients was as follows:9 kinds of medicinal materials, add of 10-fold 70% ethanol solution, soaked for 45 min, and extracted by heating reflux for 3 times,1.5 h each time.Conclusion The method can maximize extraction of ethanol dissolvable effective ingredents in 9 kinds of medicinal materials and can be used as ethonal extraction technologgary of no sugar runfei granules.
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Objective: To optimize the ethanol extraction process of Psoraleae Fructus-Myristicae Semen (psoralen-nutmeg) drug pair. Methods: Using L9(34) orthogonal design, the effects of ethanol concentration, ethanol amount, extraction time, and extraction times on the extraction process were investigated. The contents of psoralen, isopsoralen, and dehydrodiisoeugenol, dry extract yield, and total area of HPLC fingerprint characteristic peaks were used as comprehensive evaluation indexes. Results: The optimum process conditions were as follows: 50% ethanol, six times of the ethanol volume, extracted for three times, each time for 2 h. Conclusion: The method provides the basis for the determination of ethanol extraction process of Psoraleae Fructus-Myristicae Semen drug pair.
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Objective:To optimize the ethanol extraction technology for Xiaoluo granules. Methods:The extraction technology for the ethanol dissolvable ingredients in 10 kinds of medicinal materials was optimized by orthogonal experiment. The weighted sum of ba-icalin content and the dry extract rate as the index, 4 influencing factors with 3 levels each were optimized by L9 (34 ) orthogonal table. Results:The optimal ethanol extraction conditions were as follows:the medicinal material was soaked by 10-fold 50% ethanol solution for 45min, and then extracted by heating reflux for 3 times with 1. 5h each time. Conclusion: The method can effectively extract the ethanol dissolvable ingredients in 10 kinds of medicinal materials, which can be used as the ethanol extraction technology for Xiaoluo granules.
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Objective To establish the optimal ethanol extraction technology of Qingshen Granule. Methods The total content of emodin, chrysophanol and physcion, the content of tanshinoneⅡA and dry extact rate was set as indexes, orthogonal test was adopted to optimize the technology. Results The optimum extraction conditions were as follows:8 times of 80%alcohol, refluxing 3 times and 2 hours for each time. Conclusion The optimum technology of Qingshen Granule is simple, stable and effective.