文献类型：期刊文献

中文题名：基于lncRNA-mRNA共表达网络探讨党参增强衰老小鼠免疫功能的机制

英文题名：Study on the Mechanism of Codonopsis pilosula Enhancing Immune Function of Aging Mice Based on lncRNA-mRNA Co-expression Network

作者：陈冬梅[1];刘佳佳[1];蒙洁[1];康甲超[1];段永强[2];王晶[1]

第一作者：陈冬梅

机构：[1]甘肃中医药大学临床医学院,甘肃兰州730000;[2]甘肃中医药大学基础医学院,甘肃兰州730000

第一机构：甘肃中医药大学临床医学院

年份：2021

卷号：32

期号：3

起止页码：307

中文期刊名：中药新药与临床药理

外文期刊名：Traditional Chinese Drug Research and Clinical Pharmacology

收录：CSTPCD;;北大核心:【北大核心2020】；CSCD:【CSCD2021_2022】；

基金：国家自然科学基金项目(81760835,82060829);甘肃省高等学校科学研究一般项目(2018A-052)。

语种：中文

中文关键词：党参;衰老;免疫;基因芯片技术;长链非编码RNA(lncRNA);信使RNA(mRNA);小鼠

外文关键词：Codonopsis pilosula;aging;immunity;gene chip technology;long chain non-coding RNA(lncRNA);messenger RNA(mRNA);mice

摘要：目的基于对小鼠脾脏长链非编码RNA(lncRNA)、信使RNA(mRNA)表达谱的检测,探讨中药党参增强衰老小鼠免疫功能的分子机制。方法将100只昆明种小鼠随机分为对照组、模型组和党参低、中、高剂量组(5、10、15 g·kg^(-1));采用每日颈背部皮下注射D-半乳糖溶液(1.25 mg·g^(-1))建立衰老小鼠模型;党参组每日按上述剂量灌胃给药,给药体积20 mL·kg^(-1),对照组和模型组给予等量生理盐水;连续造模、给药42 d。给药结束后,测定脾脏质量和体质量,计算脾脏脏器系数;采用HE染色法对脾脏组织进行病理学观察,以透射电镜观察脾脏组织超微结构;采用基因芯片技术筛选组间差异表达的lncRNAs和mRNAs,并对差异基因进行通路富集分析;选取组间差异表达的共同lncRNAs和mRNAs进行lncRNA-mRNA共表达网络构建,并采用实时荧光定量PCR法对网络中的基因表达进行验证。结果与对照组比较,模型组小鼠的脾脏质量和脏器系数明显降低(P<0.01);脾脏组织出现病理改变,淋巴细胞发生变异、自噬及凋亡;共有96个lncRNAs和65个mRNAs在衰老后发生了显著变化;Enpp6、Cped1、Galnt15基因表达上调。与模型组比较,党参低、中、高剂量组小鼠的脾脏质量和脏器系数均明显升高(P<0.05,P<0.01);党参对衰老小鼠脾脏组织病理变化及细胞超微结构有明显改善作用;共有623个lncRNAs和435个mRNAs在高剂量党参干预后出现表达差异;差异基因的KEGG通路富集主要与免疫过程、免疫疾病相关,包括同种异体移植物排斥反应、移植物抗宿主病、自身免疫性甲状腺疾病、抗原处理及呈递等;党参高剂量组的Enpp6、Cped1、Galnt15基因表达下调(P<0.01)。结论党参对衰老小鼠脾脏具有一定的保护作用,lncRNA-mRNA共表达网络可能在党参增强衰老小鼠免疫过程中发挥重要作用。
Objective To explore the molecular mechanism of Codonopsis pilosula in enhancing the immune function spleen of mice were detected.MethodsOne hundred Kunming mice were randomly divided into control group,model group and Codonopsis pilosula low,medium and high dose groups(5,10,15 g·kg^(-1)). The aging mouse model was established by subcutaneous injection of 1.25 mg·g^(-1) D-galactose solution into the back of the neck. The Codonopsis pilosula group was given intragastric administration according to the above doses every day, and the administration volume was 20 mL·kg^(-1),while the control group and model group were given the same amount of normal saline. The above model was made continuously for 42 days. The body weight and weight of spleen were measured and the organ coefficient of spleen was calculated. The pathology of spleen was observed by HE staining and the ultrastructure of spleen tissue was observed by transmission electron microscope. The differentially expressed lncRNAs and m RNAs between groups were screened by gene chip technology and the pathway enrichment analysis ofdifferentially expressed genes was performed. The common differentially expressed lncRNAs and mRNAs betweengroups were selected for lncRNA-mRNA co-expression network construction, and Real-time fluorescencequantitative PCR was used to verify the genes in the network.ResultsCompared with the control group,the spleenweight and organ coefficient in the model group were significantly decreased(P<0.01). The spleen tissue showedpathological changes,the lymphocytes underwent mutation,autophagy and apoptosis. A total of 96 lncRNAs and65 mRNAs showed significant changes;the expression of Enpp6,Cped1,Galnt15 increased. Compared with themodel group, the spleen weights and organ coefficients in the low, medium and high dose Codonopsis pilosula groups were significantly increased(P<0.05,P<0.01). Codonopsis pilosula significantly improved the pathology andcellular ultrastructure of spleen in aging mice. The results of chip analysis showed that compared with the controlgroup, 96 lncRNAs and 65 mRNAs changed significantly after aging, and compared with the model group,623 lncRNAs and 435 mRNAs expressed differentially after high-dose Codonopsis pilosula treatment(P<0.05).KEGG pathways involved in aging and high-dose Codonopsis pilosula treatment are mainly related to immune processand immune disease,including allograft rejection,graft-versus-host disease,autoimmune thyroid disease,antigenprocessing and presentation,and so on. The expression of Enpp6,Cped1 and Galnt15 in the high-dose Codonopsis pilosula group was down-regulated compared with the model group(P<0.01).Conclusion Codonopsis pilosulahas acertain protective effect on the spleen of aging mice and the lncRNA-mRNA co-expression network may play animportant role in enhancing the immunity of aging mice.