Effect of asiaticoside on OGD/R induced injury of H9C2 cardiomyocytes based on PI3K/Akt/Beclin-1 signaling pathway / 药学学报

In order to investigate the effects of asiaticoside (Ass) on H9C2 cardiomyocytes, the present study examined the potential intervention of Ass on the proliferation and phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt)/Bcl-2 homology domain protein (Beclin-1) signaling pathway in H9C2 cardiomyocytes following oxygen and glucose deprivation/reperfusion (OGD/R) injury. H9C2 cardiomyocytes were selected as the research objects, and the activity of H9C2 was detected by cell counting kit-8 (CCK-8). H9C2 cells were divided into control group, OGD/R group, Ass low concentration group (10 μmol·L-1), Ass high concentration group (80 μmol·L-1) and Ass high concentration + chloroquine group (80 μmol·L-1 + 50 μmol·L-1). The control group was cultured under normal conditions, and the other groups were treated with oxygen and glucose deprivation for 4 h and reperfusion for 2 h. The activity and content of aspartic aminotransferase (AST), lactate dehydrogenase (LDH) and creatine kinase (CK) in the supernatant of H9C2 cardiomyocytes were detected by enzyme-linked immunosorbent assay. Autophagy staining assay kit with monodansylcadaverine (MDC) method to observe cellular autophagy; molecular docking technique to identify the molecular targets of Ass. Immunofluorescence was used to observe the effect of the drug on cell number. The expression levels of PI3K, Akt, selective autophagy adaptor protein (P62) and Beclin-1 were detected by Western blot. Compared with OGD/R group, Ass group had a protective effect from 10-80 μmol·L-1, and the activities and contents of AST, LDH and CK were decreased. The protein expression levels of PI3K, Akt, P62 and Beclin-1 were decreased. Compared with the administration group, the activities and contents of AST, LDH and CK in Ass high-concentration + chloroquine group were significantly decreased, and the protein expression levels of PI3K, Akt, Beclin-1 and P62 were significantly decreased. Immunofluorescence showed that the inhibitor group and each administration group had different degrees of protective effect compared with the model group. Asiaticoside can reduce the injury of H9C2 cardiomyocyte induced by OGD/R, reduce the content of AST, LDH and CK, reduce the expression level of P62 protein, and reduce autophagy, which may be closely related to the inhibition of PI3K/Akt/Beclin-1 signaling pathway activation.

Effects of lattice superpulsed CO 2 laser combined with asiaticoside cream ointment on repair of facial depression acne scar / 中华医学美学美容杂志

Objective:To investigate the effect of lattice superpulsed CO 2 laser combined with asiaticoside cream ointment on the repair of facial depression acne scar. Methods:A total of 124 patients with facial acne depression scar who visited the dermatology department of Zhengzhou People's Hospital from January 2019 to December 2020 were selected as subjects, including 60 males and 64 females, aged 16-38 (27.2±4.8) years. According to the random number table, the patients were divided into the control group ( n=62) and observation group ( n=62). The control group were treated with lattice superpulsed CO 2 laser, and the observation group were treated with lattice superpulsed CO 2 laser combined with asiaticoside cream ointment for 6 months. The therapeutic efficacy, Vancouver scar scale (VSS), ECCA score, skin barrier related indicators, pain duration, healing time, delayed duration and adverse reactions were compared between the two groups. Results:The total effective rate in the observation group (91.94%) was significantly higher than that in the control group (77.42%) (χ 2=5.04, P<0.05), pain duration, scab formation time, scab removal time, complete healing time, delay period and the incidence of adverse reactions were significantly lower than those in the control group [(2.76±1.04) h, (2.64±1.03) d, (6.18±1.47) d, (8.87±1.75) d, (7.89±2.16) d, 3.23% vs. (4.11±1.29) h, (3.87±1.14) d, (7.24±1.56) d, (11.05±1.93) d, (10.52±3.01) d, 12.90%, detection value = 6.42, 6.30, 3.90, 6.59, 5.59, 3.92, P<0.05]. After treatment, the VSS scale and ECCA score in the observation group were significantly lower than those in the control group (5.71±1.06, 39.12±10.64 vs. 6.42±1.17, 42.61±11.51, t=3.54, 2.26, P<0.05). After treatment, the water content of cuticle in the observation group was significantly higher than that in the control group [(40.02±14.14) vs. (34.35±11.50) AU, t=2.45, P<0.05], and transepidermal water loss, lactic acid stimulation test score and cuticle protein content were significantly lower than those in the control group [(19.07±5.70) g/(h·m 2), (2.62±1.27) score, (30.12±10.63) μg vs. (21.39±6.51) g/(h·m 2), (3.25±1.89) score, (35.10±11.19) μg, t=2.11, 2.18, 2.54, P<0.05]. Conclusions:Lattice superpulsed CO 2 laser combined with asiaticoside cream ointmentis can effectively treat acne scar and reduce adverse reactions, and the curative effect is better than single laser treatment.

Interventional effect of asiaticosdide on rats exposed to silica dust / 中华劳动卫生职业病杂志

Objective: To investigate the effect of asiaticoside for fibrosis in lung tissues of rats exposed to silica and to explore its possible mechanism. Methods: 144 SD male rats were randomly divided into control group, model group, positive drug control group, asiaticoside high-dose group, medium-dose group and low-dose group, each group included 24 rats. Rats in the control group were perfused with 1.0 ml of normal saline, and the other groups were given 1.0 ml 50 mg/ml SiO(2) suspension. Gavage of herbal was given from the next day after model establishment, once a day. Rats in the positive drug control group were administration with 30 mg/kg tetrandrine and rats in the low-dose group, medium-dose group and high-dose group were given 20 mg/kg, 40 mg/kg and 60 mg/kg asiaticoside for fibrosis respectively. Rats in the control group and the model group were given 0.9% normal saline. The rats were sacrificed in on the 14th, 28th and 56th day after intragastric administration and collect the lung tissues to detect the content of hydroxyproline, TGF-β(1) and IL-18, observe the pathological changes of the lung tissues by HE and Masson staining and determine the expressions of Col-I, a-SMA, TGF-β in lung tissues by Western Blot. Results: On the 14th day, 28th day and 56th day after model establishment, the lung tissues of rats in the model group showed obvious inflammatory response and accumulation of collagen fibers, and the degree of inflammation and fibrosis increased with time. The intervention of asiaticoside could effectively inhibit the pathological changes of lung tissues. The contents of hydroxyproline, IL-18 and TGF-β1 in lung tissues of model group were higher than those in the control group (P<0.05) , while the level of hydroxyproline, IL-18 and TGF-β1 in asiaticoside groups were significantly decreased, and the difference was statistically signicant (P<0.05) . Compared with the control group, the expression levels of Col-I, TGF-β1and α-SMA in lung tissue of model group were increased (P<0.05) , while the expression level of Col-I, TGF-β1 and α-SMA were decreased after the intervention of asiaticoside, and the difference was statistically signicant (P<0.05) . Conclusion: Asiaticoside can inhibit the increase of Col-I, TGF-β1 and α-SMA content in the SiO(2)-induced lung tissues of rats, reduce the release of TGF-β1 and IL-18 inflammatory factors in lung tissue, and then inhibit the synthesis and deposition of extracellular matrix in rat lung tissue, and improve silicosis fibrosis.

Effect of Asiaticoside on Expression of Th17/Treg Cells in DBA/1 Mice with Collagen-Induced Arthritis / 中国实验方剂学杂志

ObjectiveTo observe the effect of asiaticoside （AC） on the expression of T helper 17 （Th17） cells and regulatory T （Treg） cells in DBA/1 mice with collagen-induced arthritis （CIA）. MethodMale SPF DBA/1 mice were randomized into six groups according to body weight： control group， CIA group， methotrexate group （MTX group， ip， 0.5 mg·kg-1）， and AC low-， medium-， and high-dose groups （ig， 5， 15， 45 mg·kg-1， respectively）. Modeling was performed in rats other than the control group. To be specific， they were immunized with bovine type Ⅱ collagen and complete Freund's adjuvant on the first day and with bovine type Ⅱ collagen and incomplete Freund's adjuvant on the 21st day. Administration began on the day of the second immunization， once a day for 28 days. On the 49th day， related tissues were collected. Then， hematoxylin-eosin （HE） staining was performed to observe the pathological changes of the joints. Immunohistochemical method was used to detect the expression of interleukin-17 （IL-17） and forkhead box protein-3 （FoxP3）， the markers of Th17 and Treg cells， respectively， immunofluorescence double staining the expression of IL-17 and FoxP3 in CD4+T cells of mouse joint tissue， and flow cytometry the proportions of Th17 and Treg cells in mouse lymph nodes. ResultCompared with the control group， CIA group demonstrated joint disorder， damage of articular cartilage and bone， severe bone erosion （P<0.01）， increase in stained CD4 and IL-17 and the integral absorbance （IA） （P<0.01）， decrease in stained FoxP3 and the IA （P<0.01）， rise of Th17/Treg ratio （P<0.01）， elevation of Th17 expression in mouse lymph nodes （P<0.01）， and reduction in Treg expression （P<0.01）. Compared with CIA group， MTX group and three AC groups showed normal joints， alleviated bone erosion and damage， intact and smooth joint surface， and decrease in stained IL-17 and IA （P<0.05， P<0.01）， and MTX group and AC medium-dose and high-dose groups registered decrease in stained CD4 and IA （P<0.01） and reduction in Th17/Treg ratio （P<0.05， P<0.01）. Moreover， AC medium-dose and high-dose groups showed rise in stained FoxP3 and IA （P<0.05， P<0.01）. In the lymph nodes of mice， decrease in expression of Th17 cells （P<0.05， P<0.01） and the increase in expression of Treg cells （P<0.05， P<0.01） were observed in all the three AC group. ConclusionAC can regulate Th17/Treg balance by inhibiting the expression of Th17 cells and promoting the expression of Treg cells in CIA mice.

Preparation of asiaticoside sodium alginate repair patch and its wound healing effect / 中草药

Objective: Asiaticoside-sodium alginate repair patch (As-SA RP) was prepared and its effect on skin wound healing was investigated. Methods: Based on the cross-linking characteristics of sodium alginate and Ca2+, As-SA RP was prepared using asiaticoside as a drug model through blending with sodium alginate. The compatibility of the blend films was investigated by scanning electron microscope and FTIR. The whole layer of skin on both sides of the back spine of New Zealand's big ear rabbit was used to establish the trauma model. The repair effect of the external application of As-SA RP on skin wounds was investigated by observing the healing status and determining the expression levels of collagen (hydroxyproline as an index) and pro-inflammatory (TNF-α, IL-1β and IL-6) factors in the injured skin tissue. Results: Asiaticoside had good compatibility with sodium alginate and can be successfully prepared As-SA RP. Compared with other groups, the red swelling, bleeding, infection, and exudate on injured skin were significantly reduced, and the wound healing rate was also relatively increased after As-SA RP treatment. Moreover, the less inflammatory cells infiltration, more new capillaries and intact wound skin structure relatively tight and orderly arrangement of collagen fibers were observed in the As-SA RP group. On the other hand, the biochemical analysis showed that As-SA RP could effectively inhibit the high expression of TNF-α, IL-1β, IL-6 in the injured skin tissue and significantly increase the contents of hydroxyproline and collagen fiber. Conclusion: As-SA RP could improve the skin absorption of asiaticoside and show the better effect of skin wound healing than the active component asiaticoside.

Effect of different compatibility ratios of Centella asiatica-Rhubarb on the dissolution rate of effective components of Centella asiatica / 国际中医中药杂志

Objective@#To study the effects of different compatibility ratios of Centella asiatica- Rhubarb on the dissolution rates of madecassoside and asiaticoside.@*Methods@#By using the HPLC method to analyze the content of madecassoside and asiaticoside in the Centella asiatica extract solution as well as Centella asiatica - Rhubarb extract solution with 6 different kinds of compatibility ratios. The tests were carried out by Thermo C18 (250 mm×4.6 mm, 5 μm) by isocratic elution with acetonitrile and 2 mmol/L of beta-cyclodextrin solution as mobile phase at a flow rate of 1 ml/min, the column temperature was 30 ℃ and the detection wavelength was 205 nm.@*Results@#Compared with the Centella asiatica extract solution, the dissolution rate of madecassoside decreased significantly with the increase of the proportion of rhubarb. The combination of Centella asiatica and Rhubarb had little effect on the content of asiaticoside.@*Conclusions@#Considering the clinical application and efficacy of Centella asiatica, the optimal ratio of Centella asiatica-Rhubarb is 10:1 and 15:1.

Study on Dose-effect Relationship and Mechanism of Protective Effects of Total Asiaticoside on Gastrointestinal Motility and Enteric Nervous System in Aged Functional Dyspepsia Model Rats / 中国药房

OBJECTIVE：To explo re the dose-effect relationship and mechanism of protective effects of total asiaticoside （TA） on gastrointestinal motility and enteric nervous system （ENS）in aged functional dyspepsia （FD）model rats. METHODS ：Aged male SD rats of 16 months old were randomly divided into blank control group ，model group ，TA low dose ，medium dose and high dose groups （15，30，60 mg/kg），with 8 rats in each group. FD model was established by tail-stimulation combined with irregular diet for 4 weeks. The next day after modeling ，administration groups were given relevant doses of TA solution intragastrically ； control group and model group were given constant volume of normal saline intragastrically ，once a day ，for consecutive 15 d. Gastric emptying rate and small intestinal propulsion rate of rats were examined. ELISA were used to detect serum contents of MTL and VIP. Immunofluorescence and immunohistochemistry were proposed to measure the expression of ENS marker （S100β and GDNF）in gastric antrum tissue. The protein expression of S 100β，GFAP，PGP9.5，GDNF，p-MEK and p-ERK 1/2 in gastric antrum tissue were measured by Western blotting assay. RESULTS ：Compared with blank control group ，gastric emptying rate and small intestinal propulsion rate ，serum MTL content and protein expression of PGP 9.5 in gastric antrum tissue of model and TA low，medium dose group were decreased significantly ，while serum VIP content ，protein expressions of S 100β，GFAP，GDNF， p-MEK and p-ERK 1/2 in gastric tissue were increased significantly （P＜0.05）. Compared with model group ，gastric emptying rate and small intestinal propulsion rate of TA groups were increased significantly （P＜0.05）；except for GFAP protein in TA low dose group（P＞0.05），the serum MTL content and the expression of PGP 9.5 protein in gastric antrum tissue of rats in TA groups were increased significantly ，while serum VIP content ，protein expression of S 100β，GFAP，GDNF，p-MEK and p-ERK 1/2 in gastric antrum tissue were decreased significantly （P＜0.05）. Some or most of the content of gastrointestinal motility indexes and related factor protein expression were significantly different among TA groups （P＜0.05），and the indexes in TA high dose group could recover to the levels which were not significantly different with blank control group （P＞0.05）. CONCLUSIONS ：TA can dose-dependently improve the gastrointestinal motility deficiency and ENS dysfunction in aged FD model rats ，especially in high dose（60 mg/kg）of TA group. Its mechanism may be related with promoting the release of endogenous MTL ，inhibiting the secretion of VIP ，expression of GDNF and the activation of downstream signaling pathway ，and promoting the repair of ENS and intestinal neurons.

Elicitation effect on the production of asiaticoside and asiatic acid in shoot, callus, and cell suspension culture of Centella asiatica

Centella asiatica is an important medicinal plant which contains various phytocompounds. Asiatic acid and asiaticosideare two major compounds which are responsible for its various pharmaceutical activities. The present study analyzesthe effect of elicitor, i.e., methyl jasmonate on the synthesis of asiaticoside and asiatic acid (ATA) in shoot, callus, andcell suspension cultures of C. asiatica. A high-performance liquid chromatography analysis showed that the elicitationwith 100 µM concentration of methyl jasmonate enhanced asiaticoside content by 69-fold in callus culture, 39-fold inshoot cultures, and ATA by 1.9-fold in cell suspension culture. Thus, elicitation with methyl jasmonate is an effectivemethod of increasing the rate of biosynthesis of asiaticoside and ATA in plant cell cultures of C. asiatica

Expression Profiling Of Selected Micro Rnas Coupled With Molecular And Biochemical Analyses Of Asiaticosides In Centella Asiatica (L.)Urb In Vitro

Objective: Centella asiatica (L.) Urb from Umbelliferae is a potential source of secondary metabolites having immense medicinal value. Asiaticoside is the major therapeutic compound. In the present study, Identification of a possible relationship between concentration/transcript level expression of asiaticoside and concentrations of growth hormones at different growth stages was observed. The current study includes molecular and biochemical evaluation of stress generated in C. asiatica at different time intervals in vitro. Methods: The enhancement in auxin, cytokinin and final asiaticoside content were determined using immunoassay kits for auxin, cytokinin and HPLC analysis respectively. Transcript level expression at different growth phases was carried out using real-time RT-PCR. For isolation of stress-related miRNAs, reverse transcription of total RNA using miScript II RT Kit PCR System was carried out as per instructions. The differential expression of five selected miRNAs was done by Real-Time RT-PCR. The analysis of stress in vitro was done by quantification of Hydrogen Peroxide (H2O2), total phenolics and total antioxidants by H2O2 assay kit, total antioxidant assay kit and Folin Ciocalteau reagent respectively. The final asiaticoside content was determined by HPLC. Results: Differential expression of key genes involved in asiaticoside pathway showed significantly higher transcript expression, which is in correlation with the final asiaticoside content. The enhanced expression of miRNAs and the analysis of H2O2, total antioxidant capacity and total phenolics are suggestive of generation of oxidative stress under controlled conditions. Conclusion: The present study shows a direct correlation between oxidative stress and transcript/phytochemical estimation of asiaticoside content under in vitro conditions.

Screening of Centella asiatica drying method based on comprehensive evaluation of quality / 中草药

Objective: To explore the effects of different drying methods on the quality of medicinal materials, and screen out the optimal drying process of Centella asiatica. Methods: The whole fresh grass of C. asiatica were dried by hot air, drying in the sun, drying in the sun and hot air combined, drying in the shade, microwave and vacuum respectively. Meanwhile, the drying time and rate were determined. The characters, identification, inspection, and leachable content of C. asiatica were analyzed by the method of pharmacopoeia. The content of asiaticoside, madecassoside, asiatic acid, madecassic acid, kaempferol-3-O-tutinoside, kaempferol, and quercetin were detected by HPLC analysis; The weighted scoring method was used to sort the comprehensive evaluation of multiple indexes. Results: Different drying methods consume different time, among which drying in the sun, shade and drying at 50 ℃ for more than 100 h, and the average drying rate was 24.83%. The effects of different drying methods on the properties of medicinal materials are mainly reflected in color and odor, among which 50-70 ℃ hot air drying had a better color, which was light green, and the odor of hot air drying and microwave drying at 80 ℃ and 85 ℃ also changed significantly. Although there were some differences in moisture and ash content, both of them met the pharmacopoeia standards. The drying method also had certain effects on the leachable, the maximum was 45.70%, and the minimum content of dry extract was 29.67%. The highest content of the total active ingredient was determined by HPLC using the method of drying in the shade, which was 83.032 mg/g, and the lowest was is 75 ℃ hot air drying, which was 40.982 mg/g. The highest total content of madecassoside and asiaticoside was 80 ℃ hot air drying, and the lowest was 75 ℃ hot air drying. Weighted score in the top three of line was 70 ℃, dried at 50 ℃ after drying in the sun, hot air drying at 50 ℃, and 85 ℃ hot air drying ranked the bottom. Conclusion: In summary, the suitable drying method for the production area of C. asiatica was 70 ℃ hot air drying.

Clinical effects of Dermatix Ultra silica gel and asiaticoside cream on hyperplastic scar tissue in patients after epicanthoplasty / 中华医学美学美容杂志

Objective@#To observe the clinical efficacy of Dermatix Ultra silica gel and asiaticoside cream for the treatment or repair of scar hyperplasia which was external used in internal corner incision of patients after the internal canthus correction operation.@*Methods@#A total of 90 patients were randomly chosen and divided into three groups. All patients with epicanthus were treated with the same internal canthus correction method by one doctor. There were 30 patients in each group: 30 cases in the control group were treated without any medication, and the postoperative incision healed naturally; 30 cases were treated with Dermatix Ultra silica gel; 30 cases were treated with asiaticoside cream. Patient and observer scar assessment scale (POSAS) was used as the end points to evaluate the clinical efficacy of Dermatix Ultra silica gel and asiaticoside cream 4 weeks and 12 weeks after the operation. The satisfactory degree from all the patients were collected after 6 months follow-up and we compared the scores of three groups.@*Results@#Compared with the control group, the POSAS scores in Dermatix Ultra silica gel group and asiaticoside cream group were low (P<0.05) at 4 weeks and 12 weeks after operation. At 4 weeks after operation, the POSAS score in Dermatix Ultra silica gel group was lower than that in the asiaticoside cream group (P<0.05). At 12 weeks after operation, there was no significant difference in POSAS score between Dermatix Ultra silica gel group and asiaticoside cream group (P>0.05). Follow-up for 6 months, the satisfaction degree of patients in Dermatix Ultra silica gel group and asiaticoside cream group was higher than in the control group (P<0.05); the satisfactory degree of patients in Dermatix Ultra silica gel group was higher than that in asiaticoside cream group (P<0.05).@*Conclusions@#Dermatix Ultra silica gel and asiaticoside cream have good clinical effects in repair of scar tissue after the epicanthoplasty. Dermatix Ultra silica gel is better than asiaticoside cream in scar early improvement.

Validated high performance liquid chromatography for simultaneous determination of stability of madecassoside and asiaticoside in film forming polymeric dispersions

ABSTRACT The objective of the work was to validate the high performance liquid chromatography for simultaneous determination of stability of madecassoside and asiaticoside in Centella asiatica (L.) Urb., Apiaceae, extract-loaded film forming polymeric dispersions. High performance liquid chromatography method was validated in five topics: linearity and range, limit of detection and limit of quantitation, specificity, precision, and accuracy. Results showed the method had a good linearity (R2 > 0.9990) in the range of 5-150 µg/ml and specific. The limit of detection and limit of quantitation of madecassoside were 81 and 245 ng/ml and asiaticoside were 21 and 64 ng/ml, respectively. The percent relative standard deviation of intraday and interday precision were less than 1 and 3%, respectively. The accuracy presented as percent recovery was 101.54-103.29% for madecassoside and 100.39-102.58% for asiaticoside. This validated high performance liquid chromatography method was used to determine the stability of the formulation containing Centella asiatica extract. Centella asiatica extract-loaded film forming polymeric dispersions used Eudragit® RS 30D and Eudragit® RL 30D as film former, glycerin as plasticizer, and absolute ethanol as solvent and penetration enhancer. Three formulations with different ratio of Eudragit® RS 30D and Eudragit® RL 30D were prepared and stored for 90 days at 4 ºC, 25 ºC, and 40 ºC. Stability results showed that almost all of the formulations were unstable at 25 ºC and 40 ºC. Except, two of three formulations were stable at 4 ºC. However, the formulation was further developed to improve the stability of madecassoside and asiaticoside in the formulation.

Formulation optimization of asiaticoside loaded capsosomes by central composite design-response surface method / 中草药

Objective To fabricate the asiaticoside loaded capsosomes with CaCO3 as core (CASI) and establish the method for determination of entrapment efficiency, then the prescription of formulation and preparation process were screened with the entrapment efficiency as index. Methods An HPLC method was established to determine the contents of asiaticoside. CASI were prepared by co-precipitation method and layer-by-layer assembly technique. The encapsulation efficiency was determined by a proved centrifugation. In this study, the effect of concentrations of capsule material, pH and rotating speed on encapsulation efficiency was investigated. Results The encapsulation efficiency obtained by centrifugation was accurate and reliable. The optimized prescription was concentrations of 1 mg/mL capsule material with pH value 12, pH 7.9 of liposomes, two precursor layers, one liposome layer, rotating speed 500 r/min, and 15 min of reaction time. The CLSM images confirmed the structural integrity of the CASI. Conclusion This formulation endowed with high encapsulation efficiency, and the CASI observed by CLSM turned out to be globular shapes and was narrow in size distribution.

Study on the Tissue Distribution of Asiaticoside in SD Rats / 中国药师

Objective:To investigate the tissue distribution of asiaticoside in SD rats .Methods:The rats were injected by asiatico-side at the effective treatment dose of 42 mg kg -1 via tail vein.The rats sacrificed respectively at 5, 30 and 80 min after the injection and heart, liver, spleen, lung, kidney, stomach, intestine, brain, gonad and left thigh muscle were dissected immediately .The con-tent of asiaticoside in the tissues was determined by HPLC .Results:Asiaticoside widely distributed in SD rats , and the concentration in heart was the highest followed by liver , brain, lung, spleen and kidney .The contents in the other tissues were relatively low .Con-clusion:The method is convenient , sensitive and accurate , and can be used for the content determination of asiaticoside the tissues of SD rats.

Transdermal characteristics and mechanism of asiaticoside nanoemulsions and asiaticoside nanoemulsions-based gels / 中国中药杂志

To prepare the asiaticoside nanoemulsions (ASI-NEs) and asiaticoside nanoemulsions-based gels (ASI-NBGs), compare them with the commercial cream of asiaticoside (ASI-C) in terms of transdermal characteristics, and investigate the transdermal mechanism of ASI-NEs and ASI-NBGs. Their transdermal characteristics were studied by using Franz diffusion cells. The effect of topical ASI-NEs and ASI-NBGs on ultrastructure of rabbit skin was evaluated by using HE staining method. The localization and the permeation pathway of asiaticoside were visually investigated by using laser scanning confocal microscope (CLSM). The transdermal studies in vitro showed that the cumulative amount of ASI permeated from ASI-NEs and ASI-NBGs at 12 h after application were (3 504.30±180.93), (1 187.40±128.88) μg·cm⁻² respectively, 6.57, 2.23 times of that in the control group of ASI-C; the drug deposition of ASI-NEs and ASI-NBGs in skin was (159.48±7.47), (120.53±5.71) μg·cm⁻² respectively, 5.93, 4.48 times of that of ASI-C. HE staining of the rabbit skin after application of ASI-NEs and ASI-NBGs showed that the epidermis structure was basically intact; stratum corneum was loosed and the keratin fragment was increased; at the same time, the gap of prickle cell was increased and the basal cells were arranged loosely. The study of CLSM showed that significant percutaneous enhancer effect was observed for ASI-NEs after the topical application of 6 h, as the fluorescent compound was penetrated in the dermis and diffused uniformly. The fluorescence area and the integral optical density (IOD) were 28.81, 32.51 times of that in the FITC aqueous solution group, respectively. The fluorescent preparations showed strong fluorescence in the epidermis, but weak in deeper layers; with the increase of treatment time, the fluorescence in deeper layer was increased and stronger in skin appendages. The prepared ASI-NEs and ASI-NBGs have good transdermal characteristics and the transdermal mechanism is related to breaking the ultrastructure of stratum corneum and penetrating by the path of skin adnexa.

Inhibitory Effect of Asiaticoside on Bleomycin-induced Skin Cicatrization in Mice / 医药导报

Objective To evaluate the inhibitory effect of asiaticoside on bleomycin-induced skin cicatrization. Methods Thirty male C57BL/6 mice were randomly divided into three groups:negative control group,model control group,and asiaticoside group,ten in each group.In model control group and asiaticoside group,1 mg·mL-1bleomycin was subcutaneously injected into the dorsal skin of mice every day;4 h later,1 mL 0.9% sodium chloride solution 1 mL asiaticoside(20 mg·mL-1) was injected into the lesion skin in the model control group and the asiaticoside group,respectively.In the negative control group, the same volume of 0.9% sodium chloride solution was subcutaneously injected into the dorsal skin of the mice at the two time points every day.After 21 days,skin specimens were harvested to observe the histomorphology and detect myofibroblast proliferation and expression of inflammatory factors. Results The skin scar was significantly attenuated in the asiaticoside group as compared with the model control group,and the dermal thickness measured exhibited a gradual decrease in asiaticoside group.The expression of α-antismooth muscle antisbidy and infiltration of inflammatory cells were significantly lower in the asiaticoside group than in the model control group. Conclusion Asiaticoside inhibits the development of skin scar of mice by regulating proliferation and differentiation of myofibroblasts and down-regulating inflammatory cells.

Simultaneous Determination of Contents of Asiaticoside, Tetrahydropulmatine and Saikosaponin d in Shenji Huwei Granules by HPLC / 中国中医药信息杂志

Objective To develop an HPLC method for simultaneous determination of contents of asiaticoside, tetrahydropulmatine and saikosaponin d in Shenji Huwei Granules. Methods The analysis was performed on a R&C C18 column (250 mm × 4.6 mm, 5 μm) by using the mobile phase of acetonitrile (A) - phosphate buffer (which used potassium dihydrogen phosphate 8.34 g, potassium phosphate 0.87 g dissolved by 1000 mL water) with gradient elution (0–15 min, 20％A; 15–30 min, 20％→40％A; 30–42 min, 40％A; 42–45 min, 40％→48％A; 45–50 min, 48％A; 50–70 min, 48％→50％A). The flow rate was 1.0 mL/min; the detection wavelength was set at 210 nm; the column temperature was maintained at 30 ℃. Results Asiaticoside, tetrahydropulmatine and saikosaponin d were in the linear ranges among 0.173–2.770 μg (r=0.9999), 0.021–1.320 μg (r=0.9992), 0.151–9.660 μg (r=0.9993), respectively. The average recovery rates of asiaticoside, tetrahydropulmatine, saikosaponin d were 96.25％, 97.02％, and 97.84％, respectively, and RSD were 2.31％, 4.51％, 1.87％, respectively. Conclusion This method is simple, with good separation effect and strong specificity, and can be used for simultaneous determination of contents of asiaticoside, tetrahydropulmatine and saikosaponin d in Shenji Huwei Granules, which provides references for perfection of quality control of Shenji Huwei Granules.

Preliminary study on artificial planting technique of Centella asiatica / 中草药

Objective: To study the artificial planting technique of Centella asiatica. Methods: The yield and the total content of asiaticoside and madecassoside in C.asiatica were used as evaluating indicators. And the influences of different factors such as soil type, planting density, base fertilizer, fertilization, and shading degree on planting effect were investigated. Results: C.asiatica was suitable to be planted in peat soil/perlite (1∶1), the planting density was 20 cm × 20 cm, the base fertilizer was organic fertilizer/ compound fertilizer (1∶1) and its consumption was 7.5 g/m2, urea (18 g/m2) was applied after 4 months and compound fertilizer (45 g/m2) was applied after 5 months, the appropriate shading degree was 75%. Conclusion: The research results could be used to establish the GAP planting base of C.asiatica.

Pharmacokinetics and tissue distribution study in rats of asiaticoside-loaded modified liposomes / 中草药

Objective To prepare asiaticoside-loaded modified liposomes and to investigate the distribution and pharmacokinetics. Methods Different asiaticoside-loaded preparations (include solution, modified, and unmodified liposomes) were injected by tail vein in SD rats. HPLC method was used to detect the concentration of asiaticoside in the tissue and plasma samples. And the concentration-time profiles and pharmacokinetic parameters were then obtained and compared to get the variances. Results The concentration-time profiles of asiaticoside-loaded preparations guided along the single compartment model which the weight is 1/C2. The elimination half-life of asiaticoside solution and different asiaticoside liposomes were (14.52 ± 0.56), (101.35 ± 12.47), (149.82 ± 20.00), and (159.58 ± 16.46) min, respectively. The AUC of asiaticoside solution and different asiaticoside liposomes were (1 929.70 ± 159.00), (57 004.35 ± 8 710.89), (93 736.52 ± 12 710.76), and (64 737.48 ± 6 365.28) min∙μg/mL, respectively. The mass fraction of asiaticoside in each organ increased, especially in the pulmonary which increased from (4.94 ± 0.94) μg/g to (39.12 ± 12.04) μg/g. Conclusion The sustained release and targeting effects in SD rats were obvious of the asiaticoside-loaded modified liposomes.

Determination of madecassoside and asiaticoside in Centella asiatica formula granules by HPLC method / 药学实践杂志

Objective To establish a HPLC method for determination of madecassoside and asiaticoside in Centella asiatica formula granules.Methods Chromatographic separation was performed on Ultimate AQ-C18 column(4.6 mm×250 mm,5 μm) with mobile phase of acetonitrile(A)-2 mmol/L β-cyclodextrin(0~30 min:21% A→23% A;30~60 min:23% A→25% A).The flow rate was set at 1.0 ml/min, the column temperature at 30 ℃ and detection wavelength at 205 nm.Results Madecassoside and asiaticoside showed good linearity (r>0.9995) in the ranges of 0.187 7~3.754 μg and 0.184 3~3.686 μg respectively.The specificity, repeatability, precision,recovery and stability were satisfied to the method validation requirements of China Pharmacopoeia.Conclusion The method can determine madecassoside and asiaticoside in Centella asiatica formula granules.