Response surface methodology-based optimization of ultrasound-assisted extraction of β-sitosterol and lupeol from Astragalus atropilosus(roots) and validation by HPTLC method

Objective:To optimize the ultrasonication method for efficient extraction ofβ-sitosterol and lupeol from the roots of Astragalus atropilosus using Box-Behnken design of response surface methodology(RSM),and its validation by high performance thin layer chromatography(HPTLC)method.Methods:Ultrasonication method was used to extractβ-sitosterol and lupeol from Astragalus atropilosus(roots).RSM was used to optimize the different extraction parameters viz.liquid to solid ratio(10–14 m L/g),temperature(60-80℃)and time(40–60 min)to maximize the yield ofβ-sitosterol and lupeol.The quantitative estimation ofβ-sitosterol and lupeol was done in chloroform extract of Astragalus atropilosus by validated HPTLC method on 10 cm×20 cm glass-backed silica gel 60 F254 plate using hexane and ethyl acetate(8:2,v/v)as mobile phase.Results:A quadratic polynomial model was found to be most appropriate with regard to R1(yield of total extraction;R2/%CV=0.9948/0.28),R2(β-sitosterol yield;R2/%CV=0.9923/0.39)and R3(lupeol yield;R2/%CV=0.9942/0.97).The values of adjusted R2/predicted R2/signal to noise ratio for R1,R2,and R3 were 0.9782/0.9551/48.77,0.9904/0.9110/31.33,and 0.9927/0.9401/36.08,respectively,indicating a high degree of correlation and adequate signal.The linear correlation plot between the predicted and experimental values for R1,R2,and R3 showed high values of R2 ranging from 0.9905-0.9973.β-sitosterol and lupeol in chloroform extract of Astragalus atropilosus were detected at Rf values of 0.22 and 0.34,respectively,atλmax=518 nm.The optimized ultrasonic extraction produced 8.462%w/w of R1,0.451%w/w of R2 and 0.172%w/w of R3 at 13.5 m L/g liquid to solid ratio,78℃of temperature and 60 min of time.Conclusions:The experimental findings of RSM optimized extraction and HPTLC analysis can be further applied for the efficient extraction ofβ-sitosterol and lupeol in other species of Astragalus.

Nigella sativa oil alleviates doxorubicin-induced cardiomyopathy and neurobehavioral changes in mice:In vivo and in-silico study

Objective:To investigate the effect of Nigella sativa oil on cardiomyopathy and neurobehavioral changes induced by doxorubicin in mice.Methods:Swiss strain of albino female mice were divided into 6 groups of 5 animals in each:GroupⅠ(control group),groupⅡ(doxorubicin,10 mg/kg,i.v.),groupⅢ,Ⅳ,andⅤ(Nigella sativa oil;1.5,3,and 6 mL/kg,respectively),groupⅥ(Nigella sativa oil per se;6 mL/kg,p.o.).The duration of treatment was 15 d(10 days’pre-treatment and 5 days’post-treatment)and doxorubicin was administered on day 11th of the treatment schedule.Following Nigella sativa oil treatment,neurobehavioral tests,cardiac hypertrophy tests,and biochemical tests in serum and tissues were performed.Neurological tests included assessment of anxiety-like behavior in the elevated plus maze,spontaneous alternation behavior in the cross maze,and depression-like behavior in modified forced swim tests.Biochemical tests included serum lactate dehydrogenase and creatinine kinase-MB,malondialdehyde and reduced glutathione in tissues.Lastly,molecular docking was used to estimate the affinity of the phytoconstituents of Nigella sativa oil with histone deacetylases.Results:Nigella sativa oil treatment significantly(P<0.001)restored doxorubicin-induced neurobehavioral changes,decreased lactate dehydrogenase and creatinine kinase-MB in the plasma,malondialdehyde contents in tissues,and increased reduced glutathione level.Besides,no significant alteration was observed in Nigella sativa oil per se group as compared to the control.Molecular docking showed that Nigella sativa oil components had appreciable binding affinitiy with the protein cavities of HDAC1 and HDAC6.Conclusions:The result shows that Nigella sativa oil exerts anxiolytic,antidepressant,and memory-enhancing effects in addition to cardioprotective effect against doxorubicin-induced cardiomyopathy in mice.The modulatory effect of Nigella sativa oil on oxidative stress could contribute to the cardioprotective effect and associated neurobehavioral changes in mice.

A Review on Survivin as a Prognostic and Therapeutic Cancer Biomarker

Survivin is an antiapoptotic protein expressed in tumor cells that correlates with aggressiveness, prognosis and treatment. Objective: This review summarizes significance of survivin expression among different tumor types highlighting the potential value of survivin as an important biomarker that may have an impact on patient’s response to therapy with prognostic implications.