Anti-fatigue Effect of Water Extract from Centella asiatica(L.)Urban on Mice

[Objectives]To study the anti-fatigue effect of water extract from Centella asiatica(L.)Urban on mice.[Methods]After intragastric administration of low,medium and high concentration of water extract from Centella asiatica(L.)Urban for 14 d,the rotarod time and the content of serum lactic acid in mice were determined,respectively.[Results]Compared with the control group,the rotarod time of mice in the low and medium concentration groups was significantly prolonged(P<0.05),but there was no significant difference between the control group and the high concentration group;the content of serum lactic acid in the medium concentration group was significantly lower than that in the control group(P<0.01),but there was no significant difference between low concentration group and high concentration group and the control group.[Conclusions]A certain concentration of water extract from Centella asiatica(L.)Urban had a good anti-fatigue effect.

Molecular Identification of Alternaria alternate (Fr.) Keissl. from the Leaf Blight Disease of Centella asiatica L.

Centella asiatica (L.), frequently known as Thankuni, is an important ethnobotanical plant in Bangladesh. This study was conducted to evaluate the morphological characteristics, cultural factors and molecular identification of the causal agent of Alternaria leaf blight disease of C. asiatica. The potato dextrose agar (PDA) medium recorded the maximum mycelial growth (69 mm), followed by the yeast extract agar (YEA) medium, while the honey peptone agar (HPA) medium recorded the lowest growth (27 mm). The optimal pH and temperature for mycelial growth of Alternaria alternata were 6 and 30°C, respectively. Internal transcribed spacer (ITS) region of Alternaria alternata PCR products measured 558 bp and blast search showed 99% sequence similarity with Alternaria alternata species complex. To the best of our knowledge, Alternaria leaf blight disease caused by Alternaria alternata is the first record in Bangladesh.

Attenuation of Collagen Induced Arthritis by Centella asiatica Methanol Fraction via Modulation of Cytokines and Oxidative Stress

Objective To investigate the anti-inflammatory, antioxidant and anti-arthritic effects of Centello asiatica methanolfraction （CAME） on collagen-induced arthritis （ClA）, an animal model of rheumatoid arthritis. Methods Arthritis was induced in female wistar rats by immunization with porcine type II collagen. The CIA rats were treated orally with CaME （50, 150, and 250 mg/kg/day） for 15 d （beginning on day 21 of the experimental period）. The clinical, histological, biochemical, and immunological parameters were assessed. Results CaME treatment （150 and 250 mg/kg） significantly attenuated the severity of CIA and reduced the synovial inflammation, cartilage erosion, and bone erosion as evident from both histological and radiographic data. The escalated plasma levels of pro-inflammatory cytokines TNF-α, IL-1β, IL-6, and IL-12 alongwith nitric oxide in CIA rats decreased significantly on CaME treatment. The serum levels of type-Ⅱ collagen antibody were significantly lower in rats of CaME （150 and 250 mg/kg） treated group than those in the arthritic group. Furthermore, by inhibiting the above mediators, CaME also contributed towards the reversal of the disturbed antioxidant levels and peroxidative damage. Conclusion Our results clearly indicate that oral administration of CaME suppresses joint inflammation, cytokine expression as well as antioxidant imbalance, thereby contributing to an amelioration of arthritis severity in CIA rats.

A new triterpene and a saponin from Centella asiatica

A new triterpene and a saponin, named 2α,3β23-trihydroxyurs-20-en-28-oic acid （1） and 2α,3β,23-trihydroxyurs-20-en-28-oic acid O-α-L-rhanmopyranosyl-（1→4）-O-β-D-glucopyranosyl-（1→6）-O-β-D-glucopyranosyl ester （2）, have been isolated from the aerial part of CenteUa asiatica. Their structures were elucidated by spectral methods, including 2D-NMR spectra.

Antidiabetic effect of combination of fractionated-extracts of Andrographis paniculata and Centella asiatica: In vitro study

Objective: To examine the effect of combination of Andrographis paniculata herb fraction(AHF) and Centella asiatica herb fraction(CHF) on PPAR γ and GLUT4 mRNA expression in 3 T3-L1 adipocyte, and its effect on insulin-stimulated glucose uptake. Methods: 3 T3 L1 adipocyte cells were used to investigate gene expression of PPARy reverse transcription-polymerase chain reaction method. The γ and GLUT4 protein b adipocyte cells were differentiated by using insulin, dexamethasone and 3-isobutyl-1-methylxanthine from 3 T3-L1 cells. Pioglitazone, AHF, CHF and the combination of both herbs were evaluated on glucose uptake activity, PPAR γ and GLUT4 mRNA expressions in 3 T3-L1 adipocyte. Results: The results showed that combination of AHF at 30 μg/mL and CHF at 10 μg/mL could enhance insulin-stimulated glucose uptake. The combination also increased PPAR γ and GLUT4 mR NA expressions significantly in comparison to those of negative control(DMSO). These effects were equal in comparison to those of pioglitazone(0.02 μM) and its single extracts Conclusions: The combination of AHF and CHF can increase glucose uptake and insulin sensitivity through up-regulation of PPAR γ and GLUT4 mRNA expressions in 3 T3-L1 adipocyte.

Vegetative Growth and Molecular Identification of Fusarium equiseti Isolated from Wilt Disease of Centella asiatica L. in Bangladesh

Centella asiatica (L.) is commonly known as Thankuni plant and has ethnobotanical importance in Bangladesh. Present experiment was conceded to investigate the wilt disease of C. asiatica, vegetative growth and molecular characterization of pathogenic fungi. Pathogenic fungus, Fusarium equiseti was identified as a causal agent of wilt disease in C. asiatica. The effect of culture media on the mycelial growth of F. equiseti showed the highest (89.25 mm) on potato dextrose agar (PDA) medium followed by carrot agar (CA) medium and the lowest growth (40.25 mm) was measured in HA medium. The optimal temperature and pH for mycelial growth of F. equiseti were 30&deg;C and 7, respectively. The genetic variation of the selected species of fungi, the internal transcribed spacer (ITS) region was amplified using ITS4 and ITS5 primers and sequenced. The PCR product of the ITS region of F. equiseti was 535 bp. Phylogenetic tree of thirty-seven strains of Fusarium sp. based on the nucleotide sequences of the ITS region using the neighbor-joining method with 1000 bootstrapping indicated that 98% - 100% identity with MN886590.1 JUF0046 (F. equiseti). ITS sequences are generally constant, or show little variation within species, but vary between species in a genus. The ITS region is relatively short and can be easily amplified by PCR using universal single primer pairs. Genetic distance exhibited high level of similarity with identical ITS sequences. To date, no published research articles are found on the molecular identification of F. equiseti, the causal agent of fusarium wilt disease of C. asiatica in Bangladesh.

Compound’s Pre-Screening of <i>Withania somnifera</i>, <i>Bacopa monnieri</i>and <i>Centella asiatica</i>Extracts

Spectral fluorescence signature, Gas Chromatography-Mass Spectrometry and Liquid Chromatography-Mass Spectrometry for identification of chemical and bioactive compounds were applied to study the plant extracts of Withania somnifera, Centella asiatica and Bacopa monnieri which are related to the possible treatment of mental diseases as Alzheimer, Parkinson and Depression. These plants are known for different positive phytotherapeutic effects on the human brain without negative post-, adverse or after effects to the treated individuals, and have been recommended in several medical studies. Therefore, we selected these plants for further analysis, based on the inhibition results of in vitro Amyloid Beta fibrillation tests made by previous measurements. With this study a first screening of the complex plant extract mixtures was performed, to get an initial overview about known and unknown ingredients. In all three plants, similar main compounds were identified, however in different quality and quantity. These may provide substantial information on which compound combinations might be mainly responsible for the positive effects and should be further investigated being responsible for reducing the fibrillation process of Amyloid Beta.

Efficacy Study on the Anti-Aging Emulsion Containing Carnosine and Centella Asiatica Extract

A clinical study was conducted on the anti-aging effects of an emulsion containing carnosine and centella asiatica extract to provide a reference for the evaluation and development of anti-aging cosmetics.Sixty healthy females,aged 35~60,were divided into the test sample and placebo groups.Skin elasticity and wrinkles of the participants were analyzed before they used the emulsion and after they used it for 8 weeks.Instrument measurement results showed that after 8 weeks,skin elasticity and wrinkles in the members of the test sample group improved relative to those of the placebo group.Clinical effect assessment results showed that subjects did not experience adverse skin reactions,and their skin wrinkles showed improvement.Self-assessment results showed that the score of the total satisfaction of the test sample was 4.3,and 100%subjects are satisfied with the test sample.Emulsion containing carnosine and centella asiatica extract has a certain anti-aging effect.

Production of asiaticoside from centella(Centella asiatica L.Urban)cells in bioreactor

Objective:To investigate the effects of some culture conditions on production of asialicoside from centella(Centella asiatica L.Urban)cells cultured in 5-L bioreactor.Methods:The centell cell suspension culture was conducted in 5-L bioreactor to investigate the growth and asiaticoside accumulation under various conditions.Asiaticoside content was determined by HPLC analysis.Results:The results showed that the cell growth and asiaticoside accumulation peaked after 24d of culture at an agitation speed of 150 r/min and aeration rate of 2.5 L/min.The cell biomass reached a maximum value of 302.45 g fresh weight(31.43 g dry weight)and growth index of 3.03with inoculum size of 100 g.However,asiaticoside content was the highest(60.08 mg/g dry weight)when culture was initiated with an inoculum size of 50 g.Conclusions:The present study found the suitable conditions for growth of centella cells and their asiaticoside production in bioreactor.

The Stimulatory Effects of the Antimicrobial Agents Bavistin, Cefotaxime and Kanamycin on <i>In Vitro</i>Plant Regeneration of <i>Centella asiatica</i>(L.)—An Important Antijaundice Medicinal Plant

Antimicrobial agents such as bavistin, cefotaxime and kanamycin were evaluated for their effects on the rapid shoot regeneration from nodal explants of Centella asiatica (L.). Filter sterilized bavistin (250 mg/L) was augmented alone and in combination with cytokinins such as BAP and TDZ into the media to trace the effect on regeneration. On this basis, the potential use of bavistin (150 mg/L) along with BAP (2.0 mg/L) was evaluated which showed the maximum shoot number (6.6) and shoot length (4.4 cm) respectively. Cefotaxime at the concentration of 100 μM/L was found to be effective to obtain the maximum shoot number formation (5.8) with the regeneration frequency (90%). Kanamycin at the concentration of 80 μM/L induced maximum shoot regeneration (5.12). Kanamycin at 100 μM/L or at higher concentrations reduced the shoot regeneration. The best rooting response was noticed when in vitro regenerated microshoots were transferred to the rooting medim which was supplemented with IBA (2.0 mg/L). This combination generates 90% of regeneration frequency and maximum number of roots per shoot (14.2) and high root length (4.2 cm). The rooted plants were acclimatized and transferred to field for survivalance. The addition of antibiotics was found to be more effective and safer for using since their effects on regeneration were found to be negligible.

Comparative Efficacy of <i>Tagetes erecta</i>and <i>Centella asiatica</i>Extracts on Wound Healing in Albino Rats

The study was undertaken to evaluate the comparative efficacy of hydroalcoholic extracts of leaves of Tagetes erecta (T. erecta) and aerial parts of Centella asiatica (C. asiatica) on Excision, Incision and Dead space wound models in albino rats. Extract of T. erecta and C. asiatica (P T. erecta extract (250 and 500 mg/kg) showed significantly increased the wound breaking strength in incision wound model and wet and dry granulation tissue weights, breaking strength in a dead space wound model compare to control and C. asiatica treated group (P T. erecta extract showed potent wound healing activity then the reported C. asiatica in different wound parameters.

Decreasing a-synuclein aggregation by methanolic extract of Centella asiatica in zebrafish Parkinson's model

Objective: To observe the effects of Centella asiatica(C. asiatica) methanolic extract on a-synuclein aggregation and its expression in rotenone-exposed zebra fish.Methods: Zebra fish(Danio rerio) were exposed to 5 m g/L rotenone for 28 days and coincubated with 2.5, 5.0 and 10.0 m g/mL of C. asiatica methanolic extract. The medium was changed every 48 h for maintain the concentration of rotenone and extract. After 28 days zebra fish were sacrificed on the ice block and protein was isolated from zebra fish brain for ELISA of dopamine and Western blotting of a-synuclein. Immunohistochemistry was conducted to observe the a-synuclein expressions from histopathological preparation of zebra fish brain. The head were soaked in 10% formaline for less than 24 h and embedded onto paraffin block, then sliced for immunohistochemistry using anti a-synuclein antibody. We also measured zebra fish motility for 5 min in each week.Results: C. asiatica has important bioactive compounds such as asiaticoside that has antiin flammatory and antioxidant properties. It may inhibit cascade reaction due to oxidative stress induced by rotenone. Decreasing reactive oxygen species proposed probability of radical attack to a-synuclein protein that caused aggregation and increase of its expression.The motility of zebra fish was also maintained in C. asiatica groups due to the increasing dopamine level in rotenone-induced zebra fish. High level of reactive oxygen species inactivated enzyme for dopamine synthesis such as tyrosine hydroxylase, and oxidized dopamine itself. Oxidized dopamine increased a-synuclein aggregation. Thus, the dopamine level decreased in rotenone-induced zebra fish, but C. asiatica increased dopamine level.Conclusions: C. asiatica has a potential to be developed as an anti-Parkinson's disease treatment due to its capability for minimized the sign of Parkinson's such as a-synuclein aggregation and expression, increasing motility and dopamine as well.

Comparative study of standardized Centella asiatica and combination of levodopa and benzerazid HCl on Parkinson′s model zebrafish

OBJECTIVE To investigate the motility,dopamine,serotonin and pro-BDNF level of Centella asiatica(CA)comparable to the combination of levodopa and benzerazid HCL on zebrafish model Parkinson′s.METHODS Rotenone 5μg·L-1 induced to adult zebrafish(8months)for 28 dto made zebrafish model Parkinson′s.CA concentration were used 5 and 10μg·mL-1.The combination of levodopa 100 g and 25 mg benzerazid(L-DOPA agonist)was given at 1 μg·mL-1 at the same time to rotenone,and CA as well.Motility assessment conducted for every 7dstarted on day 0until day 28.Five fish were subjected in the 2L tank(25cm×16.5cm×12cm).Three vertical lines were drawn on the tank at equal distances,dividing the tank into four zones(the length of each zone was 6.25cm).Locomotor activity was measured for 5min by counting the number of lines that adult zebrafish crossed.Fish were sacrificed by decapitating on ice water.Dopamine level measured from whole brain by ELISA,serotonin and pro-BDNF by immunoreactivity at substantia nigra.RESULTS The locomotor activity of rotenone treated fish were significantly decrease starts at 7 dcompared to control group.Interestingly on 10μg·mL-1 group there are increasing motility start at day 7 and slightly decrease until day 28,but on L-DOPA group increasing motility at day 7 followed by decreasing motility significantly(P<0.05)until day 28.Dopamine level of rotenone group decreased compared to control group,CA 10μg·mL-1 and L-DOPA group increased significantly to rotenone group which CAμg·mL-1 higher than L-DOPA group.Serotonin and pro-BDNF level had the same profile.Its significantly increased on rotenone group compared to control group.Serotonin on group with CA administration significantly decreased compared to rotenone group,while rotenone plus CA 10μg·mL-1 had no significant differet to L-DOPA group(P>0.05).Pro-BDNF on rotenone with CA 5μg·mL-1 had no significant difference,while compare to rotenone with CA 10-1 μg·mLgroup and L-DOPA group were significantly different(P<0.05).CONCLUSION Rotenone produces reliable Parkinson′s zebrafish model by decreasing motility and dopamine level.CA was more stable,increasing motility than L-DOPA administration.CA and L-DOPA increased dopamine level,but decreasing serotonin and pro-BDNF.

Enzymatic pretreatment and microwave extraction of asiaticoside from Centella asiatica

The extraction of asiaticoside from Centella asiatica by enzymatic pretreatment and microwave extraction (EPME) was studied in this article. The effects of several important factors such as temperature of enzymatic pretreatment, liquid to solid ratio and microwave radiation time were investigated by quadric regression orthogonal design experiment and were analyzed by response surface. An extraction model with well forecast performance was then established. The results indicate that the optimum extraction condition was as follows: liquid to solid ratio was 36mL/g, temperature of enzymatic pretreatment was 45℃, enzymatic time was 30min, and microwave ra-diation time was

Prophylactic effects of asiaticoside-based standardized extract of Centella asiatica(L.) Urban leaves on experimental migraine: Involvement of 5HT1A/1B receptors

The present study aimed at evaluation of prophylactic efficacy and possible mechanisms of asiaticoside(AS) based standardized extract of Centella asiatica(L.) Urban leaves(INDCA) in animal models of migraine. The effects of oral and intranasal(i.n.) pretreatment of INDCA(acute and 7-days subacute) were evaluated against nitroglycerine(NTG, 10 mg·kg-1, i.p.) and bradykinin(BK, 10 μg, intra-arterial) induced hyperalgesia in rats. Tail flick latencies(from 0 to 240 min) post-NTG treatment and the number of vocalizations post-BK treatment were recorded as a measure of hyperalgesia. Separate groups of rats for negative(Normal) and positive(sumatriptan, 42 mg·kg-1, s.c.) controls were included. The interaction of INDCA with selective 5-HT1 A, 5-HT1 B, and 5-HT1 D receptor antagonists(NAN-190, Isamoltane hemifumarate, and BRL-15572 respectively) against NTG-induced hyperalgesia was also evaluated. Acute and sub-acute pre-treatment of INDCA [10 and 30 mg·kg-1(oral) and 100 μg/rat(i.n.) showed significant anti-nociception activity, and reversal of the NTG-induced hyperalgesia and brain 5-HT concentration decline. Oral pre-treatment with INDCA(30 mg·kg-1, 7 d) showed significant reduction in the number of vocalization. The anti-nociceptive effects of INDCA were blocked by 5-HT1 A and 5-HT1 B but not 5-HT1 D receptor antagonists. In conclusion, INDCA demonstrated promising anti-nociceptive effects in animal models of migraine, probably through 5-HT1A/1B medicated action.

LED光质对积雪草生长及总苷含量的影响

通过研究LED光质对水培积雪草生长和生理指标的影响,探索积雪草在植物工厂中适宜的光质,为积雪草在植物工厂水培过程中的光环境适应机制和高产优质提供理论依据。本试验以未生根的积雪草幼苗为材料进行水培,利用植物工厂的LED人工光源,设计蓝光(B)、红蓝光组合(R∶B=2∶8、R∶B=3∶7、R∶B=5∶5、R∶B=7∶3、R∶B=8∶2)、红光(R)等7种不同光质的处理,测定各处理水培积雪草的生长指标、光合色素及有效成分的含量和产量。试验结果表明:R有利于水培积雪草的分枝数、叶片数、叶面积、匍匐茎、全株鲜质量和全株干质量;R∶B=2∶8、R∶B=3∶7、B有利于水培积雪草的叶绿素a、总叶绿素和类胡萝卜素的积累;R∶B=7∶3有利于积雪草苷和羟基积雪草苷含量的形成与积累;R有利于积雪草总苷产量积累。

复方积雪草组方联合中药结肠透析对CKD4~5期患者的临床疗效及肠道菌群的影响

目的:观察复方积雪草组方联合中药结肠透析治疗CKD4~5期患者的临床疗效及对肠道菌群的影响。方法:将48例CKD4~5期非透析且属肾虚血瘀浊毒证的患者随机分为两组,在基础治疗上,治疗组予复方积雪草组方并联合中药结肠透析;对照组予酪酸梭菌活菌片,治疗12周,观察两组治疗前后的中医证候积分,肾小球滤过率(eGFR),血肌酐(Scr),尿素氮(BUN),血尿酸(UA),血白蛋白(Alb),总胆固醇(TC)等指标,同时粪便标本行细菌16S扩增子测序分析肠道菌群组成及丰度。结果:与治疗前相比,治疗组的Scr及BUN水平未见明显上升(P>0.05);而对照组的Scr及BUN水平均明显升高(P<0.01),两组的eGFR变化率差异有统计学意义(P<0.05)。治疗后,治疗组的Alb水平明显升高,TC水平明显下降;UA水平、中医证候积分显著低于治疗前及对照组,差异均有统计学意义(P<0.05~0.01)。治疗组的临床总有效率81.8%;对照组的总有效率42.3%;两组间比较差异有统计学意义(P<0.01)。两组治疗前优势菌门构成有厚壁菌门、拟杆菌门、变形菌门,两组在治疗前后优势菌群组成均有变化,但差异无统计学意义(P>0.05)。但两组间治疗后菌群组成比较,差异有统计学意义(P<0.05)。结论:通过复方积雪草组方联合中药结肠透析干预后,患者的Scr及eGFR水平稳定,Alb上升,UA及TC水平、中医证候积分较治疗前有明显改善,部分有益菌群的丰度增加。

中医外治综合疗法对痤疮皮损修复作用的临床研究初探

目的观察中医外治综合疗法治疗寻常痤疮皮损的效果。方法选取60例寻常痤疮患者,随机分为治疗组和对照组。治疗组采用锋勾针排脓放血后滚针导入中药药液治疗,对照组采用口服丹参酮胶囊配合克林霉素磷酸酯外用溶液外涂治疗。比较两组治疗后痤疮等级、皮损软硬度、疗效、满意度、安全性及复发率。结果治疗后,治疗组痤疮等级、皮损软硬度、皮损修复疗效、满意度均优于对照组(P<0.05)。两组不良反应无统计学差异(P>0.05)。治疗组复发率小于对照组(P<0.05)。结论相比丹参酮口服结合克林美溶液外涂治疗,中医外治综合疗法对寻常痤疮皮损的修复能力更强,疗效更好。