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目的检测16周龄MRL/lpr小鼠发病情况,并研究发病后的MRL/lpr小鼠脾脏自噬水平。方法采用雌性MRL/lpr小鼠和雌性C57BL/6小鼠,分别于16周龄时检测2组小鼠脏器系数,ELISA检测血清抗ds-DNA、ANA的水平,淋巴结、肾脏病理HE染色观察,以检测MRL/lpr小鼠发病情况。取发病小鼠,Western blot检测2组小鼠脾脏自噬相关蛋白Beclin-1、ATG5、P62、LC3Ⅱ/Ⅰ的表达,Real-time PCR检测自噬相关基因Beclin-1、ATG5、Atg12、LC3B mRNA的表达,流式检测2组小鼠脾脏中B细胞和浆细胞的比例。结果 16周龄MRL/lpr小鼠脾脏、淋巴结脏器系数升高,血清中自身抗体水平显著升高,淋巴结和肾脏均出现一定程度的病理性改变。此时脾脏中的自噬相关蛋白Beclin-1、ATG5、LC3Ⅱ/Ⅰ表达均高于C57BL/6小鼠,P62的表达显著低于C57BL/6小鼠,自噬相关基因ATG5 mRNA的表达也显著升高。此外,相较于C57BL/6小鼠,MRL/lpr小鼠脾脏中B细胞数量减少,而抗体分泌型浆细胞数量显著升高。结论 16周龄MRL/lpr小鼠已出现狼疮样症状,出现自噬水平异常升高的现象,并且脾脏中抗体分泌型浆细胞的增多可能与异常升高的自噬水平密切相关。
Objective To detect the incidence of 16-week-old MRL / lpr mice and study the splenic autophagy in MRL / lpr mice after the onset of disease. Methods The MRL / lpr mice and female C57BL / 6 mice were used to detect the organ coefficient of the two groups at 16 weeks of age. Serum levels of anti-dsDNA and ANA were measured by ELISA. Lymph nodes and renal pathology were observed by HE staining To detect the incidence of MRL / lpr mice. The expression of Beclin-1, ATG5, P62 and LC3Ⅱ / Ⅰ were detected by Western blot and the expressions of autophagy-related genes Beclin-1, ATG5, Atg12 and LC3B mRNA The proportions of B cells and plasma cells in the spleens of two groups of mice were detected by flow cytometry. Results In 16-week-old MRL / lpr mice, the spleen and lymph node organ coefficient increased, the levels of serum autoantibodies increased significantly, and some pathological changes occurred in lymph nodes and kidneys. The expression of autophagy-related proteins Beclin-1, ATG5 and LC3Ⅱ / Ⅰ in spleen was higher than that in C57BL / 6 mice, the expression of P62 was significantly lower than that in C57BL / 6 mice, and the expression of autophagy-related gene ATG5 mRNA was also significant Rise. In addition, the number of B cells in the spleens of MRL / lpr mice was reduced and the number of antibody-secreting plasma cells was significantly increased compared to C57BL / 6 mice. Conclusions 16-week-old MRL / lpr mice have developed lupus-like symptoms with an abnormally elevated level of autophagy. Moreover, the increase of antibody secreting plasma cells in the spleen may be closely related to abnormally elevated autophagy.