CAJ | 학술논문

目的：观察华蟾素注射液对小鼠脾淋巴细胞增殖、活化的影响，探讨其对免疫系统调节的作用，为阐明其免疫作用机制提供药理学依据。方法无菌分离小鼠脾淋巴细胞，制备淋巴细胞悬液，按临床给药量设置华蟾素注射液不同浓度组给药。MTT法分别检测刀豆蛋白A（ ConA）刺激和脂多糖（ LPS）刺激的小鼠脾淋巴细胞增殖；用华蟾素注射液预孵育淋巴细胞6 h，分别加入Con A或LPS诱导淋巴细胞活化，利用流式细胞仪通过Aimplex?对细胞上清液进行GM-CSF、TNF-α，IFN-γ，IL-1α，IL-2，IL-6，IL-10细胞因子检测，评价其免疫效果。结果华蟾素注射液7.82 mg/ml、15.63 mg/ml和31.25 mg/ml浓度组与ConA协同刺激淋巴细胞增殖。1.96 mg/ml和62.50 mg/ml浓度组与LPS协同抑制细胞增殖。华蟾素注射液可促进TNF-α和IL-6分泌，一定程度抑制IFN-γ和GM-CSF分泌，与不同刺激剂作用可不同程度促进或抑制IL-2、IL-1α和IL-10的分泌。结论华蟾素注射液可一定程度促进B淋巴细胞增殖，但对体液免疫的作用效果和作用机制还有待进一步研究。
목적：관찰화섬소주사액대소서비림파세포증식、활화적영향，탐토기대면역계통조절적작용，위천명기면역작용궤제제공약이학의거。방법무균분리소서비림파세포，제비림파세포현액，안림상급약량설치화섬소주사액불동농도조급약。MTT법분별검측도두단백A（ ConA）자격화지다당（ LPS）자격적소서비림파세포증식；용화섬소주사액예부육림파세포6 h，분별가입Con A혹LPS유도림파세포활화，이용류식세포의통과Aimplex?대세포상청액진행GM-CSF、TNF-α，IFN-γ，IL-1α，IL-2，IL-6，IL-10세포인자검측，평개기면역효과。결과화섬소주사액7.82 mg/ml、15.63 mg/ml화31.25 mg/ml농도조여ConA협동자격림파세포증식。1.96 mg/ml화62.50 mg/ml농도조여LPS협동억제세포증식。화섬소주사액가촉진TNF-α화IL-6분비，일정정도억제IFN-γ화GM-CSF분비，여불동자격제작용가불동정도촉진혹억제IL-2、IL-1α화IL-10적분비。결론화섬소주사액가일정정도촉진B림파세포증식，단대체액면역적작용효과화작용궤제환유대진일보연구。
Objective To analyze the effect of cinobufacini injection( Cinobufacini)on mice spleen lymphocyte proliferation and activa_tion in vitro. To discuss its role in regulation of immune system,provide the pharmacological evidence to clarify its immune mechanism. Methods Aseptic isolated the mice spleen lymphocyte,which prepared the lymphocyte suspension,according to the clinical dosage. The Cinobufacini group was administered with different concentration. MTT method was used to detect the concanavalin A(ConA)and lipopolysaccharide(LPS) stimulating mice spleen lymphocyte proliferation separately. Cinobufacini pre-incubate the lymphocytes 6 h,add ConA or LPS separately to in_duce the lymphocyte activation. Theg flow cytometry with Aimplex?kit on cell supernatant was used to detect the cytokines of GM-CSF,TNF-α,IFN-γ,IL-1α,IL-2,IL-6 and IL-10,evaluating its immunological effect. Results The Cinobufacini concentrations were 7. 82 mg/ml,15. 63 mg/ml and 31. 25 mg/ml and ConA co-stimulated the lymphocyte proliferation. 1. 96 mg/ml,62. 50 mg/ml groups and LPS co inhib_ited the proliferation. Cinobufacini can promote the secretion of TNF-αand IL-6,inhibit the IFN-γand GM-CSF secretion in a certain de_gree. The different stimulating agents can promote or inhibit IL-2,IL-1αand IL-10 secretion in different degree. Conclusion Cinobufacini can promote the proliferation of B cells in a certain extent,but the effect and mechanism of humoral immunity remains to be further studied.