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Objective To establish and verify the design space for the extraction process of Carthami Flos (CF) based on the concept of quality by design (QbD). Methods The safflower was used as a model drug. The critical evaluation indicators were determined through literature and previous research experience. Fishbone diagram and Failure Mode Effect Analysis (FMEA) were carried out to determine the critical process parameters (CPPs). The mathematical models of CPPs and critical evaluation indicators were established using the Box-Behnken experimental design method. Results The critical evaluation indicators were the extraction amount of total flavonoid, hydroxysafflor yellow A (HSYA), and total solids. The water-adding amount, extraction temperature, extraction time, and extraction times were determined as the CPPs by the fishbone diagram and FMEA. The variance analysis results of Box-Behnken experiments showed that the P values of established models were less than 0.000 1, indicating that the models have good prediction ability. The recommended operating space were as follows: the extraction times were twice and 2.5 h for each time, the water-adding amount was 23.5-25 mL/g crude drug and the temperature was 65-71 ℃. Conclusion The establishing of design space for CF extraction improves the correlation between the extraction process parameters and extract quality, which provides a reference for the applicability of the design space approach in the field of CMM.
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Objective To observe the protective effect of hydroxysafflor yellow A(HSYA) on anoxia/reoxygenation (A/R) injury of neonatal primary cardiomyocytes, and its relationship with phosphoinositide 3-kinase/protein ki-nase B/glycogen synthase kinase 3β(PI3K/Akt/GSK3β) signaling pathway. Methods Primary cardiomyocytes of neonatal rats were isolated from the rats and incubated for 48 hours. The cells were adhered to each other and then divided into five groups:control group (Con group), anoxia/reoxygenation group (A/R group),HSYA treatment group(A/R+H group),PI3K inhibitor (LY294002)treatment group(A/R+L group)and HSYA+LY294002 treat-ment group (A/R+H+L group),then to collect the supernatant fluid of each group to measure LDH.The flow cy-tometry was used to measure the apoptotic cells. The protein levels of Bcl-2,Bax,Akt,p-Akt (Ser473),GSK3β, p-GSK3β (Ser9) were evalated by Western blot. Results A/R increased LDH release,the apoptosis rate (P<0.001),and the expression of pro-apoptotic protein Bax (P <0.001) with the decrease of anti-apoptotic protein Bcl-2,p-Akt(Ser473), p-GSK3β(Ser9)(P<0.001) as compared with the control group. HSYA treatment de-creased LDH release,the apoptosis rate (P<0.001),and the expression of Bax (P<0.001) and increase the ex-pression of Bcl-2,p-Akt(Ser473),p-GSK3β(Ser9)(P<0.001). Compared with the A/R+H group,the expres-sion of Bax was increased (P<0.001),while the expression of Bcl-2, p-Akt(Ser473), p-GSK3β(Ser9)was de-creased (P<0.001) in the A/R+H+L group. Conclusions HSYA protects rats'cardiomyocytes from anoxia/reoxy-genation injury by regulating PI3K/Akt/GSK3β signaling pathway.
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To investigate the pharmacokinetic characteristics of active constituents of Guhong injection in rats with cerebral ischemia reperfusion injury. The middle cerebral artery occlusion (MCAO) model was established in our studies, and then all the rats received iv administration of Guhong injection (2.1 mL·kg⁻¹). The blood concentrations of aceglutamide and hydroxysafflor yellow A (HSYA) were determined by high performance liquid chromatography (HPLC) method at different time points. The concentration-time curves were drawn and pharmacokinetic data were obtained by DAS 3.2.6 software. The results showed that aceglutamide and HSYA showed good linear relationship within the ranges of 1.5-500 mg·L⁻¹ (R²=0.997 5) and 0.33-40 mg·L⁻¹ (R²=0.998 9) respectively. This quantitative method showed a high recovery rate, good precision and stability. The main pharmacokinetics parameters of t1/2α, t1/2β, CL₁, CL₂, AUC0-t, AUC0-∞, Vd1, and Vd2 were (0.139±0.007) and (0.155±0.017) h, (0.803±0.046) and (2.233±0.410) h, (0.016±0) and (0.149±0.018) L·h⁻¹·kg⁻¹, (0.015±0.001) and (0.446±0.016) L·h⁻¹·kg⁻¹, (133.335±3.844) and (9.298±0.179) mg·h·L⁻¹, (143.851±3.595) and (14.464±1.451) mg·h·L⁻¹, (0.009±0.001) and (0.223±0.007) L·kg⁻¹, (0.006±0.001) and (0.212±0.032) L·kg⁻¹, respectively. The results showed that the established HPLC method was highly specific, and could be used for the simultaneous detection of aceglutamide and HSYA of Guhong injection in MCAO rats, which was conducive to pharmacokinetic studies. Pharmacokinetic data and parameters could provide reference for continuous administration and interval administration of the drug.
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Objective: To investigate the enhancing effect of Borneolum Syntheticum and Acori Talarinowii Rhizoma on penetrating blood-brain barrier (BBB) of hydroxysafflor yellow A (HSYA) in rat. Methods: The concentration of HSYA in rat plasma and brain was determinated after ig administration of 20.00 mg/kg of HSYA to rats with or without Borneolum Syntheticum and Acori Talarinowii Rhizoma. And then, the character of penetrating BBB through the result of the AUCbrain/AUCbloodwas evaluated. Results: The AUC 0-8h, were (77 228.76±2 873.19), (81 949.04±2 283.11), and (28 479.63±2 431.71) ng·mL-1·min for blood and (55 925.0± 2 434.28), (82 768.53 ±3 277.00), and (70 914.29 ±2 900.71) ng·mL-1·min for brain of the control group, Borneolum Syntheticum group, and Acori Talarinowii Rhizoma group. The AUCbrain/AUCblood, were 0.72, 1.01, and 2.49, respectively. Conclusion: Borneolum Syntheticum and Acori Talarinowii Rhizoma do enhance the penetrating BBB of HSYA (P<0.05). Further more, Acori Talarinowii Rhizoma could affect the distribution of HSYA in rats.