[Immuno-regulatory effect of 3'-meisoindigo in mice of various germlines].
OBJECTIVE
To investigate the effect of 3'-meisoindigo on the proliferation and the biological function of the splenocyte and thymocyte of mouse, which were 8 weeks old masculinity BALB/c, C57BL/6 and F1 hybridization mouse.


METHODS
Cells of thymus and spleen were harvested and prepared as the unicell suspension, then treated with 5, 10, 15, 20, 25 micromol/L 3'-meisoindigo. The cell proliferation was by MTT method, concentration of IL-12 was dectected by ELISA method, the mRNA levels of Bcl-2 and CDK2 were decected by RT-PCR. The cell cycle, apoptosis ratio, death ratio and intracellular ROS concentration were detected by FCM method. The protein level of Bcl-2, CDK2 and Bax were detected by immumofluorescence method.


RESULTS
15, 20, 25 micromol/L 3'-meisoindigo can inhibit the proliferation of thymocyte and splenocyte (P < 0.05). It had dose-dependent and time-dependent manner. 3'-meisoindigo inhibit the secretion of IL-12, even at 5 micromol/L concentration. 15 micromol/L 3'-meisoindigo decrease the mRNA level of Bcl-2 and CDK2, induced apoptosis and G2 arrestting of the thymocyte and splenocyte. (P < 0.05). The intracellular ROS level increased after treated by 3'-meisoindigo at 15 micromol/L for 24 h (P < 0.05). There were no difference among three germ line mouse.


CONCLUSIONS
Above 15 micromol/L, 3'-meisoindigo can inhibit the proliferation and externalization function of thymocyte and splenocyte from different germ line mouse, meanwhile the mRNA and protein level of Bcl-2 and CDK2 decrease, the Bax protein expressed increased, the intracellular ROS level increase too.
Hematology Department, The Second Affiliated Hospital of SUN Yat-Sen University, Guangzhou 510102, China. wxj6209@163.com
Xiu-Ju
XJ
Wang
Song-Mei
SM
Yin
Li-Ping
LP
Ma
Da-Nian
DN
Nie
Shuang-Feng
SF
Xie
Yi-Qing
YQ
Li
D000276:Adjuvants, Immunologic / Q000494:pharmacology*; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D051357:Cyclin-Dependent Kinase 2 / Q000235:genetics / Q000378:metabolism; D007211:Indoles / Q000008:administration & dosage / Q000494:pharmacology; D018664:Interleukin-12 / Q000378:metabolism; D031219:Isatis / Q000737:chemistry; D008214:Lymphocytes / Q000187:drug effects / Q000378:metabolism; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D010936:Plant Extracts / Q000008:administration & dosage / Q000494:pharmacology*; D028502:Polygonum / Q000737:chemistry; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D013045:Species Specificity; D013154:Spleen / Q000166:cytology* / Q000276:immunology; D013950:Thymus Gland / Q000166:cytology* / Q000276:immunology
D000276:Adjuvants, Immunologic; D007211:Indoles; D010936:Plant Extracts; D019253:Proto-Oncogene Proteins c-bcl-2; D012333:RNA, Messenger; D018664:Interleukin-12; C047330:N-methylisoindigotin; D051357:Cyclin-Dependent Kinase 2