RESUMO
OBJECTIVE@#The present study aimed to evaluate the therapeutic effect and explore the underlying mechanisms of Longxue Tongluo Capsule (LTC) on ischemic stroke rats.@*METHODS@#Twenty-six rats were randomly divided into four groups, including sham group, sham + LTC group, MCAO group, and MCAO + LTC group. Ischemic stroke rats were simulated by middle cerebral artery occlusion (MCAO), and LTC treatment group were orally administrated with 300 mg/kg of LTC once daily for seven consecutive days. LTC therapy was validated in terms of neurobehavioral abnormality evaluation, cerebral infarct area, and histological assessments. The plasma metabolome comparisons amongst different groups were conducted by UHPLC-Q Exactive MS in combination with subsequent multivariate statistical analysis, aiming to finding the molecules in respond to the surgery or LTC treatment.@*RESULTS@#Intragastric administration of LTC significantly decreased not only the neurobehavioral abnormality scores but also the cerebral infarct area of MCAO rats. The interstitial edema, atrophy, and pyknosis of glial and neuronal cells occurred in the infarcted area, core area, and marginal area of cerebral cortex were improved after LTC treatment. A total of 13 potential biomarkers were observed, and Youden index of 11 biomarkers such as LysoPC, SM, and PE were more than 0.7, which were involved in neuroprotective process. The correlation and pathway analysis showed that LTC was beneficial to ischemic stroke rats via regulating glycerophospholipid and sphingolipid metabolism, together with nicotinate and nicotinamide metabolism. Heatmap and ternary analysis indicated the synergistic effect of carbohydrates and lipids may be induced by flavonoid intake from LTC.@*CONCLUSION@#The present study could provide evidence that metabolomics, as systematic approach, revealed its capacity to evaluate the holistic efficacy of TCM, and investigate the molecular mechanism underlying the clinical treatment of LTC on ischemic stroke.
RESUMO
In order to obtain salt-tolerant calli of Dandelion (Taraxacum officinale Weber), calli were induced from leaf explants of Dandelion on Murashige and Skoog's medium supplemented with 2.0 mg/L 6-benzyladenine and 0.5 mg/L 2,4-dichlorophen oxyacetic acid With 1.5% NaCl as selection pressure, most calli became brown and dead, whereas some new cell clusters appeared at the edge of the brown calli after 2 to 3 weeks. The survived cells were picked out and sub-cultured every 3 weeks onto the fresh selection medium and salt-tolerant calli were finally obtained through 4 continuous selections on the selection medium supplemented with 1.5% NaCl. Salt-tolerant calli increased steadily under a fixed NaCl stress though their relative growth rate decreased with increased NaCl concentration whereas the control calli which were sub-cultured by 4 continus selections on salt free medium ceased to grow under the same condition. This result indicated that the salt-tolerance of the selected calli is improved and this character is stable. Compared with the control, the SDS-PAGE pattern of the salt-tolerant calli had a unique 34 kD protein band. Its 30 kD and 18 kD protein bands were up-regulated. Further more, within the NaCl stress range up to 1.5%, the activities of antioxidant enzymes such as super oxidase dimutase, peroxidase and catalase, and the proline contents of the salt-tolerant calli were higher than those of the control. The results indicated that the selected calli with improved and stable salt tolerance were cell variants. The accumulation of the organic compatible solutes including proteins and the enhanced antioxidant capabilities in the salt tolerant calli are the two ways for them to regulate their osmotic homeostasis and alleviate the secondary reactive oxygen spexies damage respectively.