ABSTRACT
Objective@#Comparison of effects of Zingiberis Rhizoma and Aristolochia manshuriensis on diuretic effect and acute renal injury in rats by combining two methods of co-decoction and mixed-decoction. The effects of in vitro observation on normal human renal tubular epithelial cells (HK-2) were observed.@*Methods@#The rats were randomly divided into five groups: blank group, positive control group, aristolochia manshuriensis group, combined decoction group and divided decoction group, 8 rats in each group. The water-loading rat model was established by intragastric administration of normal saline. The urine of rats was collected and the volumes of urine were measured for 24 hours after the corresponding drugs were given to each group. After 2 weeks of gavage of the corresponding drugs in each group, the serum BUN, SCr and urine UCr and PRO levels were measured by 7600P automatic biochemical analyzer, and renal histopathology were observed by HE staining. The HK-2 cells were cultured in vitro and divided into control group, Aristolochia manshuriensis group, mixed decoction group and sub-decoction group. After 24 hour intervention, the activity of cells was detected by CCK-8 method and the apoptosis was observed by Hoechst stain method.@*Results@#There was no significant difference in 24 hour urine output between the groups (P>0.05). Compared with Aristolochia manshuriensis group, the kidney coefficient (0.010 1 ± 0.005 8 vs. 0.013 3 ± 0.007 8), SCr (38.52 ± 0.58 μmol/L vs. 46.61 ± 0.72 μmol/L), BUN (8.55 ± 0.12 mmol/L vs. 10.21 ± 0.30 mmol/L), UCr (52.21 ± 0.89 μmol/L vs. 57.71 ± 0.67 μmol/L), PRO (29.89 ± 0.18 mg/L vs. 34.23 ± 6.05 mg/L) of combined decoction group significantly decreased (P<0.05). The survival rate of HK-2 cells (72.45% ± 3.70% vs. 55.92% ± 8.39%) in combined decoction group significantly increased (P<0.01), and the apoptosis rate (7.9% ± 2.6% vs. 31.6% ± 9.1%) significantly decreased (P<0.01).@*Conclusions@#The traditional co-decoction method of Aristolochia manshuriensis compatibility with Zingiberis Rhizoma can achieve a certain attenuation effect, and the mixed-decoction group can not achieve the attenuating effec.
ABSTRACT
Objective Comparison of effects of Zingiberis Rhizoma and Aristolochia manshuriensis on diuretic effect and acute renal injury in rats by combining two methods of co-decoction and mixed-decoction. The effects of in vitro observation on normal human renal tubular epithelial cells (HK-2) were observed. Methods The rats were randomly divided into five groups: blank group, positive control group, aristolochia manshuriensis group, combined decoction group and divided decoction group, 8 rats in each group. The water-loading rat model was established by intragastric administration of normal saline. The urine of rats was collected and the volumes of urine were measured for 24 hours after the corresponding drugs were given to each group. After 2 weeks of gavage of the corresponding drugs in each group, the serum BUN, SCr and urine UCr and PRO levels were measured by 7600P automatic biochemical analyzer, and renal histopathology were observed by HE staining. The HK-2 cells were cultured in vitro and divided into control group, Aristolochia manshuriensis group, mixed decoction group and sub-decoction group. After 24 hour intervention, the activity of cells was detected by CCK-8 method and the apoptosis was observed by Hoechst stain method. Results There was no significant difference in 24 hour urine output between the groups (P>0.05). Compared with Aristolochia manshuriensis group, the kidney coefficient (0.010 1 ±0.005 8 vs. 0.013 3 ± 0.007 8), SCr (38.52 ± 0.58 μmol/L vs. 46.61 ± 0.72 μmol/L), BUN (8.55 ± 0.12 mmol/L vs. 10.21 ± 0.30 mmol/L), UCr (52.21 ± 0.89 μmol/L vs. 57.71 ± 0.67 μmol/L), PRO (29.89 ± 0.18 mg/L vs. 34.23 ± 6.05 mg/L) of combined decoction group significantly decreased (P<0.05). The survival rate of HK-2 cells (72.45% ± 3.70% vs. 55.92% ± 8.39%) in combined decoction group significantly increased (P<0.01), and the apoptosis rate (7.9% ± 2.6% vs. 31.6% ± 9.1%) significantly decreased (P<0.01). Conclusions The traditional co-decoction method of Aristolochia manshuriensis compatibility with Zingiberis Rhizoma can achieve a certain attenuation effect, and the mixed-decoction group can not achieve the attenuating effec.
ABSTRACT
OBJECTIVE:To establish a method for simultaneous determination of hyperoside,quercitrin,luteoloside, kaempferol,quercetin,rutin,luteolin and isorhamnetin in total flavanones of Sedum sarmentosum Bunge. METHODS:UPLC-MS/MS method was adopted. The determination was performed on ZOBAX SB C18column with mobile phase consisted of methanol-5 mmol/L ammonium formate aqueous solution(45:55,V/V)at the flow rate of 0.4 mL/min. The column temperature was 30 ℃, and sample size was 2 μL. The electrospray ionization source(ESI)was used;ion source temperature was 400 ℃;desolvation temperature was 300 ℃;desolvation gas flow was 600 L/h;capillary voltage was 3 000 V;nebuliser pressure was 45 psi;the work mode was multiple reaction monitoring mode;detection mode was negative ion mode. The established method was used to determine the contents of 8 components in 3 batches of total flavanones of Sedum sarmentosum Bunge. RESULTS:The linear ranges of hyperoside,quercitrin,luteoloside,kaempferol,quercetin,rutin,luteolin and isorhamnetin were 10.0-640.0,0.5-32.0, 4.5-288.0,8.0-512.0,50.0-3 200.0,2.0-128.0,12.5-800.0 and 25.2-1 612.8 ng/mL(r≥0.991 4),respectively. The limits of detection were 5.0,0.25,2.25,4.0,25.0,1.0,6.25 and 12.6 ng/mL,respectively.The limits of quantitation were 10.0,0.5,4.5, 8.0,50.0,2.0,12.5 and 25.2 ng/mL,separately. RSDs of precision,stability(24 h)and reproducibility tests were no more than 4.3%(n=6). The recoveries were 95.9%-100.6%,and RSDs were 1.5%-3.8%(n=6). The contents of hyperoside,quercitrin, luteoloside,kaempferol,quercetin,rutin,luteolin and isorhamnetin in 3 batches of total flavanones of Sedum sarmentosum Bunge were 507.88-560.37,42.95-50.36,63.52-71.80,1 695.10-1 753.27,10 569.28-10 612.99,25.76-30.13,2 795.22-2 877.43 and 4 869.55-4 971.30 μg/g,respectively. CONCLUSIONS:The established method can be used for simultaneous determination of 8 components in total flavanones of Sedum sarmentosum Bunge.