1.Preparation and characterization of rabbit anti-mouse zona pellucida 2 antibodies
Chuncheng NIE ; Guanying YAO ; Zuowu CAO
Journal of Southern Medical University 2015;(4):522-525,548
Objective To prepare rabbit anti- mouse zona pellucida 2 (mZP2) polyclonal antibodies and test their immunoactivity. Methods Recombinant proteins of mZP2 expressed in Rosetta transformant was separated by SDS-PAGE, and the gel strips containing the recombinant mZP2 were cut out and emulsified to immunize New Zealand white rabbits. The antibody response of the antiserum was detected by ELISA, and the specificity of the antiserum was verified by immunohistochemical assay. The effect of the antiserum on the binding of oocytes with acrosomal reacted sperm was tested by sperm-egg binding assay. Results ELISA results showed that the immunized rabbit produced anti-mZP2 antiserum. The antiserum reacted specifically with the zona pellucida of mouse ovarian sections. Sperm-egg binding assay showed that treatment of the oocytes with the anti-mZP2 antiserum caused decreased binding of zona pellucida with the acrosomal reacted sperm by 43.7%. Conclusion We obtained rabbit anti-mouse ZP2 polyclonal antibodies that can inhibit the binding of oocytes with acrosomal reacted sperm.
2.Preparation and characterization of rabbit anti-mouse zona pellucida 2 antibodies
Chuncheng NIE ; Guanying YAO ; Zuowu CAO
Journal of Southern Medical University 2015;(4):522-525,548
Objective To prepare rabbit anti- mouse zona pellucida 2 (mZP2) polyclonal antibodies and test their immunoactivity. Methods Recombinant proteins of mZP2 expressed in Rosetta transformant was separated by SDS-PAGE, and the gel strips containing the recombinant mZP2 were cut out and emulsified to immunize New Zealand white rabbits. The antibody response of the antiserum was detected by ELISA, and the specificity of the antiserum was verified by immunohistochemical assay. The effect of the antiserum on the binding of oocytes with acrosomal reacted sperm was tested by sperm-egg binding assay. Results ELISA results showed that the immunized rabbit produced anti-mZP2 antiserum. The antiserum reacted specifically with the zona pellucida of mouse ovarian sections. Sperm-egg binding assay showed that treatment of the oocytes with the anti-mZP2 antiserum caused decreased binding of zona pellucida with the acrosomal reacted sperm by 43.7%. Conclusion We obtained rabbit anti-mouse ZP2 polyclonal antibodies that can inhibit the binding of oocytes with acrosomal reacted sperm.
3.Effects of FAR on myocardial fibrosis of diabetic rats
Dongdong WANG ; Sumei HE ; Guanying ZHANG ; Di YIN ; Xin HUANG ; Lijuan CHEN ; Xiao CHEN ; Tong WEI ; Qunli WEI ; Huankai YAO
Chinese Pharmacological Bulletin 2015;(4):509-513
Aim To study the effects of Free Anthra-quinone from Rhubarb (FAR)on myocardial CTGF and collagen expression and interstitial fibrosis in dia-betic rats.Methods The male SD rats were randomly divided into normal group (CON),diabetic cardiomy-opathy group (DCM) and FAR treatment group (FAR).Streptozocin was intraperitoneally injected in-to the animals in the latter 2 groups to induce diabetic rat model.The model was expected to be stable for 2 weeks before the treatment.At the end of the 8th week in treatment,fasting plasma glucose and heart mass in-dex were measured.Masson staining was used to ob-serve the myocardial fibrosis.RT-PCR was used to de-tect the mRNA levels of CTGF,procollagen type Ⅰand collagen type Ⅲ.Immunohistochemical method was used to detect the content of CTGF.ELISA was used to detect the depositions of collagen type I and collagen type Ⅲ. Results Compared with CON group,fasting plasma glucose,heart mass index,the degree of myocardial fibrosis,and the expressions of CTGF,collagen type I and collagen type Ⅲ in left ven-tricular myocardial tissue of DCM group were signifi-cantly increased. However, compared with DCM group,fasting plasma glucose,heart mass index,the degree of myocardial fibrosis,and the expressions of CTGF,collagen type I and collagen type Ⅲ in left ven-tricular myocardial tissue of FAR-treated rats were sig-nificantly decreased.Conclusion FAR retards the process of myocardial fibrosis in diabetic rats by down-regulating the expression of CTGF,reducing the syn-thesis and depositions of collagen type I and collagen type Ⅲ.