Phytochemical Studies, Antimicrobial and Anti-Inflammatory Properties of the Hydroethanolic Extract of Kalanchoe pinnata (Lam.) Pers. from Togolese Flora

Kalanchoe pinnata (Lam.) Pers. is known as a plant that has many special benefits such as anti-inflammatory and antibacterial. The present study was carried out to perform a phytochemicals study and evaluate the antimicrobial and anti-inflammatory activity of the hydroethanolic extract of Kalanchoe pinnata (Lam.) Pers. leaves. After phytochemicals screening, the content of phenolic compounds, proanthocyanidol and flavonoids in the extract of this plant was determined spectrophotometrically. Antimicrobial activity was assessed using the micro-dilution technique on 96-well plates in liquid medium, combined with agar spreading. Anti-inflammatory activity was assessed using the 1% carrageenan induced rat paw oedema model. Phytochemical screening revealed the presence of alkaloids, saponins, triterpenes and sterols, phenols and flavonoids in the plant extract in varying proportions. The extract contained (0.049 ± 0.03 µg EAG/mg extract) total polyphenols, (0.215 ± 0.025 µg CE/mg extract) proanthocyanidins and (385.435 ± 0.0328 µg ER/mg ES) flavonoids. The hydroethanolic extract of the leaves of this plant inhibited the in vitro growth of the microbial strains studied to varying degrees. The MIC of the extract varied from 12.5 to 25 mg/mL and the BMC from 12.5 to 50 mg/mL. The plant did not show any activity on 1% carrageenan-induced rat paw edema.

Nutraceutical with Anti-Inflammatory Activity for the Management of Airway Remodeling in Bronchial Asthma: <i>Kalanchoe integra</i>Var. Crenata (Andr.) Cuf Leaf Extract

Background: Kalanchoe integra is widely used in folklore medicine as an antiasthmatic agent. Previous studies have shown the ameliorating effect of Kalanchoe integra leaf extract [KILE] on bronchial hyperesponsiveness and inflammation. Further, the stabilizing effect of Kalanchoe sp on mast cell degranulation, suggests that Kalanchoe species are suitable candidates for allergic asthma therapy. This study is designed to investigate the anti-asthmatic potential of KILE and monitor the accompanying histopathological and immunobiochemical changes that occur in an animal model of bronchial asthma using ovalbumin sensitized guinea pigs. Method: Thirty male guinea pigs were divided into five groups of six animals each. Bronchial asthma was simulated in guinea pigs using ovalbumin. Both low dose (300 mg/kg) and high dose extract (900 mg/kg) were administered daily for 42 days. Prednisolone (2.5 mg/kg) was the standard drug used. Results: Guinea pigs in all KILE treated groups maintained the integrity of their airway structures: bronchial folds and walls, alveoli, alveolar ducts and sacs. KILE and prednisolone caused a reduction in immune parameters (p 0.001), extent of bronchoconstriction, bronchial wall thickness and goblet cell accumulation in the sensitized guinea pigs. Conclusion: This study demonstrates the anti-asthmatic potential of KILE during prolonged administration by the oral route.

The Effect of Wide-Range Photosynthetic Active Radiations on Photosynthesis,Growth and Flowering of Rosa sp.and Kalanchoe blossfeldiana

Miniature roses (Rosa sp.) and Kalanchoe blossfeldiana were grown at photon flux densities (PFD) ranging from 60 to 670 μmol·m-2·s-1 (associated with a temperature gradient from 20.0°C to 24.0°C [TEMP1]) and from 50 to 370μmol·m-2-s-1 (associated with a temperature gradient from 22.5°C to 26.5°C [TEMP2]). The experiment was conducted in a greenhouse compartment at latitude 59° north in mid-winter. The daily photosynthetic active radiations (PAR) ranged from 4.3 to 48.2 and 3.6 to 26.6 mol·m-2·day-1 in the TEMP1 and TEMP2 treatments, respectively. Time until flowering in miniature roses decreased from about 50 to 35 days in the TEMP1 treatment and from 50 to 25 days in the TEMP2 treatment, when the PFD increased from 50 to 370μmol·m-2·s-1. In Kalanchoe time until flowering was decreased to the same extent (about 15 days) in both temperature treatments when PFD increased from 50 to 370 μmol·m-2·s-1. The number of flowers and the plant dry weight in miniature roses increased up to 300 – 400 μmol·m-2·s-1 PFD (21.6 - 28.8 mol·m-2 day-1 PAR), while flower stem fresh weight and plant dry weight in Kalanchoe increased up to 200 – 300 μmol·m-2·s-1 at TEMP1. Measurements of the diurnal carbon dioxide exchange rates (CER) in daylight in small plant stands of roses in summertime showed that CER was saturated at about 300 μmol·m-2·s-1 PFD at 370 μmol·mol-1 CO2 and at 400 – 500 μmol·m-2·s-1 PFD at 800 μmol·mol-1 CO2. For Kalanchoe similar results were obtained. Increasing the CO2 concentration from 370 to 800 μmol·mol-1 increased the CER in roses (48%) as well in Kalanchoe (69%). It was concluded that 15 to 20 mol·m-2·day-1 combined with about 24°C air temperature and high CO2 concentration will give a very good growth with lot of flowers within a short production time in miniature roses. For Kalanchoe 10 to 15 mol·m-2·day-1 combined with about 20°C and high CO2 produced a similar result.

Hypoglycemic and hypocholesterolemic activities of the aqueous preparation of Kalanchoe pinnata leaves in streptozotocin-induced diabetic rats

Objective:To evaluate the hypoglycemic and hypocholesterolemic activities of the aqueous preparation of Kalanchoe pinnata(K.pinnata) leaves in streplozotocin-induced diabetic rats.Methods:Diabetes mellitus was induced in rats by a single administration of streptozotocin(60 mg/Kg).Diabetic rats were then treated with aqueous K.pinnata for 30 d.Serum glucose,proteins.lipid composition,liver and kidney function indices,inflammatory markers,and key enzymes of hepatic carbohydrate and lipid metabolism were determined.Results:The untreated and treated diabetic groups lost weight and consumed less food compared to the normal group.We noted 37.9%decrease in fasting blood glucose in the treated diabetic group compared to 13.2%and 17.0%increases in normal and untreated diabetic groups respectively.Serum cholesterol and triglyceride levels were significantly(P<0.05) reduced in the treated diabetic group compared to the untreated diabetic group.Blood urea nitrogen was significantly(P<0.05) elevated in the untreated and treated diabetic groups compared lo the normal group.Serum alkaline phosphatase and hepatic pyruvate kinase activities were significantly(P<0.05) elevated in the treated diabetic group.Scrum albumin level was signilieantiy(P<0.05) reduced in the untreated diabetic group.Serum IL-6 was significantly(P<0.05) depressed in the treated diabetic group.Conclusions:The observed decrease in body weight,blood glucose and cholesterol level suggests that the aqueous K.pinnata preparation consumption may be beneficial in the management of diabetes mellitus.The observed adverse effect on alkaline phosphatase activity may be due to the combined effect of streptozotocin-induced diabetes and K.pinnata preparation administration.

Agrobacterium–mediated Transformation of Kalanchoe laxiflora

Plants with crassulacean acid metabolism(CAM) generally utilize water 20%–80% more efficiently than non-CAM plants. The whole genomes of several CAM plants have been sequenced or are being sequenced. For effective genome characterization and genome editing of CAM plants,an efficient transformation system is essential. In this study, we developed an Agrobacterium-mediated transformation protocol for Kalanchoe laxiflora, an obligate CAM plant,by optimizing several factors affecting the transformation efficiency. Agrobacterium strains AGL1, C58, EHA105,and GV3101 were all suitable for K. laxiflora transformation. Fifty-nine percent of the leaf explants yielded kanamycin-resistant and GUS-positive shoots. Polymerase chain reaction and quantitative real-time PCR(qRT-PCR) using gus A-, gus Plus-, npt II-and hpt-specific primers confirmed that the transgenes were integrated into K. laxiflora genome and expressed. This efficient transformation system will allow effective functional characterization of genes through over-or down-expression, knockout, or genome editing.

Evaluation of the analgesic,sedative-anxiolytic,cytotoxic and thrombolytic potentials of the different extracts of Kalanchoe pinnata leaves

Objective:To evaluate the analgesic,neuropharmacological,cytotoxic and thrombolytic potentials of the aqueous,ethanol and ethyl acetate extracts of Kalanchoe pinnata leaves.Methods:At the dose of 400 mg/kg body weight,the analgesic activity of the extracts were evaluated by the acetic acid-induced writhing and formalin-induced persistent pain tests while neuropharmacological activity was evaluated by the open field,hole cross and elevated plus maze tests.The cytotoxic potential was observed by brine shrimp lethality bioassay and the thrombolytic potential was investigated by clot lysis test.Results:The aqueous extract significantly suppressed the number of writhing(96.78%)as well as the formalin-induced persistent pain on the early phase(46.92%)and on the late phase(40.98%).Again in case of hole cross and open field tests,the locomotor activity was decreased significantly(P<0.001)mostly by the ethyl acetate extract.Furthermore,the sedative-anxiolytic activity was supported by the increased percent(P<0.01)of frequency into the open arm on elevated plus maze test.Besides,the extracts showed moderate lethality and thrombolytic activity.Conclusions:The findings showed that activities are comparable to the standards and in some cases are stronger than the standards.Therefore,based on the results,it is evident that it has great analgesic and sedative-anxiolytic activity with moderate cytotoxic and thrombolytic potential.

Chemical constituents from Kalanchoe hybrida and their cytotoxicity

Objective：Kalanchoe hybrida（Crassulaceae）is naturalized throughout all the island of Taiwan in China.The preliminary bioassay-guided fractionation of the crude extract of K.hybrida exhibited that the chloroform and n-butanol fractions possessed potent cytotoxicity against MCF-7,NCI-H460,and SF-268 tumor cell lines at 50μg/m L concentration.Therefore,K.hybrida was selected as a target and the chemical constituents from the chloroform and n-butanol fractions of the crude extracts of K.hybrida were identified.The potential constituents were examined for their cytotoxicity against the tumor cell lines.Methods：A combination of conventional chromatographic techniques was performed on the crude extract of K.hybrida.The chemical structures of the purified constituents were identified on the basis of spectroscopic and spectrometric analysis.Results：The purification results had led to the characterization of totally 37 compounds.The isolated compounds 1,2,and 4–12 were examined for their cytotoxicity in vitro,and bufadienolides 4–8 and flavonol glycoside 11 displayed significant cytotoxicity towards all the tested tumor cell lines among these tested compounds.Conclusion：The results indicated that these principles should be responsible for the bioactivity of corresponding partial fractions.The potential constituents could be further investigated to explore the new natural lead drugs.

基于离体模式体系的长寿花脱毒苗培养及瓶内开花

以长寿花的顶芽为外植体,通过愈伤组织诱导获得无菌苗,并进行瓶内花芽诱导研究。结果表明,长寿花茎尖最佳灭菌时间为0.1mg/L的氯化汞处理3min,成活率达到40%;愈伤组织诱导的最佳茎尖大小为0.3mm;茎尖诱导愈伤组织产生的最佳培养基为MS+2mg/L6-BA+0.3mg/LNAA+0.2mg/L2,4-D;愈伤组织诱导丛芽增殖的最佳培养基为MS+0.3mg/LNAA+1mg/L6-BA,诱导花芽分化的最佳激素配比为MS+0.1mg/LNAA+0.25mg/L6-BA+0.6mg/LGA3,花芽诱导率最高,为45.32%;瓶内开花的最佳培养基为MS(1/3NH4NO3),开花率为45%。本试验通过愈伤组织的诱导,芽的增殖,获得脱毒苗并诱导使其在瓶内开花,可为其优良品种的开发利用和快速繁殖提供技术支持。

Effect of 5'-deletion of Arabi-dopsis profilin2 promoter on its vascular specific expression

Based on the published sequence of profilin2 promoter of Arabidopsis thaliana, a full-length promoter (1667 bp) was amplified by PCR. The 5’ -end deletion fragments with length of 1380, 1153, 969 and 597 bp were then fused with gus (uid\) gene respectively. Constructed plant expression vectors were individually transferred into Kalan-choe laciniata and transgenic plants regenerated. GUS his-tochemical assay confirmed that the full-length promoter Pfnl.7 was vascular-specific. Deletion assays showed that profilin2 promoter could be divided into three parts. Deletion of fragment 1 ( -1667 -1380 bp) resulted in constitutive expression, suggesting that element(s) responsible for vascular-specific expression might exist in this region. Fragment 2 located at -1153 - -597 bp strongly inhibited gus gene expression. Fragment 3 ( -597 - -1 bp) is considered as a basic domain of profilin2.