ABSTRACT
Objective To explore the impact of different doses of butylphthalide on renal interstitial fibrosis in mice with obstructive nephropathy, and to discuss the correlation between Nrf-2 expression level and renal interstitial fibrosis. Methods Totally 72 male CD-1 mice of clean grade were selected and randomly assigned into 4 groups:sham operation group (Sham), model group (UUO), low dose of butylphthalide group (NBPL) and high dose of butylphthalide group (NBPH). In addition to the control group, UUO model was established in other three groups. NBPL group was given NBP 150 mg/(kg· d) by gavage since postoperative day 1,NBPH group was given NBP 220 mg/(kg·d) by gavage. Sham group and UUO group were given equal volume of saline by gavage. Six mice were sacrificed at the third, 7th, 14th day, respectively. The obstructive renal tissue was selected for immunohistochemical staining and Western blot assay. The expression levels of Nrf- 2 and type Ⅰ collagen were detected by Western blot assay. Results The IOD value of type Ⅰ collagen was increased with time in UUO group, and which showed a gradual decreasing trend in Sham group, NBPL group and NBPH group (P<0.05). The IOD values at different time points were significantly higher in UUO group, NBPL group and NBPH group compared to those of Sham group. The IOD values were significantly decreased in NBPL and NBPH groups than that of UUO group, and IOD value was lower in NBPH group than that of NBPL group (P<0.05). Western blot assay showed that IOD values of Nrf-2 and type Ⅰ collagen at 7 d and 14 d were increased in UUO group, NBPL group and NBPH group compared with those of Sham group. The IOD values of Nrf-2 protein were increased, and IOD values of type Ⅰcollagen were decreased, in NBPL group and NBPH group than those of UUO group. The IOD value of Nrf-2 protein was decreased in NBPL group than that of NBPH group, but the IOD value of type Ⅰcollagen was increased in NBPL group than that of NBPH group (P<0.05). Conclusion NBP can improve renal interstitial fibrosis in UUO mice, which may be related with the increased expression of Nrf-2 and the down-regulated expression of type I collagen.
ABSTRACT
Objective To develop a new method for Tinospora sagittata species identification and genetic map construction. Methods Sequence-related amplified polymorphism (SRAP) was applied for T. sagittata to carry on PCR amplification of its DNA and optimize the reaction parameter grade by grade. Results The stable and reproducible SRAP reaction system of T. sagittata has been developed. Conclusion SRAP is an effective method for T. sagittata identification in molecular degree and it has set up a foundation for the further species identification and genetic map construction of T. sagittata.
ABSTRACT
Objective: A method for determination of glycyrrhizine and paeoniflorin in Fubao Danggui Jiao by RP HPLC was established.Methods: Shim pack CLC ODS column was used with TBAH solution (10→300) methanol (36∶64) (pH=6.0) as a mobile phase for glycyrrhizine and acetonitrile water (containing 0.1% phosphoric acid) (15∶85) as a mobile phase for paeoniflorin. The detection wavelength was at 254nm and 230nm, respectively. The flow rate was 1mL?min -1 .Results: A good linearity was found at the concentration range of 100 1000ng for glycyrrhizine and paeoniflorin. The average recovery was 98.37%,97.44% and the relative standard deviation was 0.32%,0.48%, respectively.Conclusion: This method was simple,accurate,rapid,sensitive and reproducible. The RP HPLC could be used for quality control of Fubao Danggui Jiao preparations.