主 办:北 京 中 医 药 大 学
ISSN 1006-2157 CN 11-3574/R

JOURNAL OF BEIJING UNIVERSITY OF TRADITIONAL CHINESE MEDICINE ›› 2021, Vol. 44 ›› Issue (4): 332-339.doi: 10.3969/j.issn.1006-2157.2021.04.008

• Chinese Medicinal Pharmacology • Previous Articles     Next Articles

Effects of Zuogui Jiangtang Shuxin formula medicated plasma on pyroptosis and apoptosis induced by ox-LDL in J774A.1 cells*

Yang Jinwei1, Yu Rong2,3*, Wu Yongjun4#, Xiao Fan5, Liu Xiu3, Deng Yihui1   

  1. 1 College of Integrated Traditional Chinese and Western Medicine, Hunan University of Chinese Medicine, Hunan 410208, China;
    2 Graduate School, Hunan University of Chinese Medicine, Hunan 410208, China;
    3 Key Laboratory of Colleges and Universities, Traditional Chinese Medicine Prescription and Transformation of Medical Laboratory, Hunan 410208, China;
    4 School of Pharmacy, Hunan University of Chinese Medicine, Hunan 410208, China;
    5 School of International Education, Hunan 410208,China
  • Received:2020-09-27 Published:2021-04-29
  • Contact: A/Prof.Wu Yongjun, Master Supervisor. School of Pharmacy, Hunan University of Chinese Medicine, No.300,Xueshi Road, Hanpu Science and Education Park, Yuelu District, Hunan 410208. E-mail: 1463636969@qq.com
  • Supported by:
    National Natural Science Foundation of China (No.81573956, 82074400); National Key Research and Development Plan of China(No.2018YFC1704300); Key Research and Development projects of Hunan Province (No.2020SK2101); Research Foundation of Education Bureau of Hunan Province (No.18A220, 20K094); Open Fund for Traditional Chinese Medicine of Hunan Province (No.2018ZYX44).
    Ethical review: Committee of Ethical Review on Experimental Animals in Hunan University of Chinese Medicine (No.LL2018112501).

Abstract: Objective To investigate the effects of Zuogui Jiangtang Shuxin (kidney-supplementing glucose-lowering heart-soothing) formula medicated plasma on ox-LDL-induced pyroptosis and apoptosis and to explore new targets in the treatmert of diabetic atherosclerosis with this formula. Methods SD rats were administered intragastrically with 7.2 g/kg Zuogui Jiangtang Shuxin formula or 0.83 g/kg metformin combined with 12.45 mg/kg atorvastatin calcium to prepare drug-containing plasma. Zuogui Jiangtang Shuxin formula medicated plasma was then extracted for J774A.1 cells treatment. J774A.1 cells were pretreated with ox-LDL to establish the experimental model of pyroptosis and apoptosis. CCK-8 essay was used to meausre the effect of medicated plasma or ox-LDL on proliferation of J774A.1 cells and to determine the optimal concentration. J774A.1 cells were randomly divided into 4 groups: normal group, model group, Zuogui Jiangtang Shuxin medicated plasma group and Western medicine combination (metformin+atorvastatin calcium) medicated plasma group. After inducting and administration for 24 hours, LDH releasing test was used to evaluate the cell membrane integrity. Cell apoptosis were measured by using flow cytometry. The expression of NLRP3, Caspase-1,GSDMD, Caspase-3 and Bax were detected by using Western blot assay. Results 10% medicated plasma and blank plasma showed no inhibition of cells, the concetnration of which was chosen to be suitable for experiment. The cell survival rate decreased when J774A.1 cells were treated with 100 mg/L ox-LDL for the induction concentration. Compared with the blank group, 100 mg/L ox-LDL suppressed the growth of J774A.1 cells. It was also shown with LDH releasing test that LDH activity and apoptosis rate increased in the model group cells significantly(P<0.01). Moreover, ox-LDL could up-regulate the expression of NLRP3, Caspase-1,GSDMD, Caspase-3 and Bax in the model group(P<0.01). Compared with the model group, Zuogui Jiangtang Shuxin medicated plasma reduced inhibition rate, LDH release and the rate of apoptosis (P<0.01). Meanwhile, the expression of pyroptosis related protein (NLRP3, Caspase-1 and GSDMD) and apoptosis related protein Caspase-3 and Bax expression were significantly reduced(P<0.01). These effects were significantly superior to Western medicine combination group (P<0.01). Conclusion Zuogui Jiangtang Shuxin formula medicated plasma can maintain the function of J774A.1 cells, and its mechanism may be related to the inhibition of pyroptosis and apoptosis induced by ox-LDL.

Key words: Zuogui Jiangtang Shuxin formula, medicated plasma, J774A.1 cells, pyroptosis, apoptosis

CLC Number: 

  • R285.5