结核分枝杆菌感染环境中不同细胞因子对胸膜间皮细胞增殖和凋亡的影响

Effects of different cytokines on proliferation and apoptosis of pleural mesothelial cells in humanMycobacterium tuberculosis infection

目的探讨结核感染时不同细胞因子[IL-22、IL-17、γ干扰素（IFN-γ）]对胸膜间皮细胞（PMC）增殖和凋亡的影响。方法应用流式细胞术检测PMC表面细胞因子受体IL-22R、IL-17R和IFN-γR1的表达；检测IL-22、IL-17、IFN-γ以及IFN-γ与IL-22或IL-17组合对PMC中的Ki-67表达和凋亡的影响；检测结核性胸腔积液（TPE）及其分别与抗IL-22、IL-17、IFN-γ单抗组合对PMC中的Ki-67表达的影响。结果（1）结核分枝杆菌感染时PMC表面高表达IL-22R、IL-17R和IFN-γR1[PMC的阳性比例分别为（86．2±2．9）％、（41．5±4．4）％和（64．9±5．8）％]；（2）IL-22、IL-17组Ki-67阳性PMC细胞百分数分别为（31．5±2．0）％、（26．1±2．4）％，高于培养基组的（14．6±0．7）％（q值分别为6．8和4．9，均P〈0．05）；IFN-γ组为（5．2±1．2）％，低于完全培养基组（q=5．0，P〈0．05）；而IFN-γ＋IL-22组为（23．4±1．7）％，IFN-γ＋IL-17组为（21．8±3．8）％，均高于IFN-γ组（q值分别为7．3和6．7，均P〈0．05）。单独加入TPE或加入含有同型对照IgG的TPE组分别为（63±9）％、（63±11）％，均高于完全培养基组（q值分别为19．6和19．7，均P〈0．05）；TPE加入抗IFN-γ单抗之后为（82±4）％，高于TPE＋IgG组（q=7．5，P〈0．05）；TPE加入抗IL-22单抗后为（34±3）％，低于TPE＋IgG组（q=11．8，P〈0．05）；TPE加抗IL一17单抗组为（58±5）％，与TPE＋IgC组差异无统计学意义（q=2．1，P〉0．05）。（3）IFN-γ组PMC凋亡率为（19．3±1．1）％，高于完全培养基组（4．3±0．6）％（q=33．4，P〈0．05）；IL-22、IL-17组凋亡率分别为（3．8±0．6）％和（5．7±0．8）％，与完全培养基组差异无统计学意义（q值分别为1．3和3．0，均P〉0．05）。IFN-γ＋IL-22组凋亡率为（6．5±0．7）％，IFN-γ＋IL-17组为（8．7±1．7）％，均低于IFN-γ组（q值分别为28．5和23．6，均P〈0．05）。结论IL-22和IL-17促进PMC增殖而不导致其凋亡，并且能逆转IFN-γ对PMC增殖的抑制作用，以及逆转IFN-γ对PMC凋亡的诱导作用；结核分枝杆菌感染环境可促进PMC增殖，该效应可能与TPE中IL-22、IL-17和IFN-γ的综合作用有关。

Objective To investigate the effects of different cytokines （IL-22, IL-17, IFN-γ） on proliferation and apoptosis of human pleural mesothelial cells （ PMC ） during Mycobacterium tuberculosis infection. Methods The expressions of IL-22R, IL-17R and IFN-γR1 on PMC purified from tuberculous pleural effusion （TPE） were determined by flow cytometry. The effects of one or more of IL-22, IL-17, and IFN-γ on Ki-67 expression and apoptosis of PMC were explored. Ki-67 expression of PMC under the conditions of the absence or presence of exogenous TPE and with a combination of control IgG, or anti-IL- 22, -IL-17 or -IFN-γ mAbs were deternfined. Results （ 1 ） During Mycobacterium tuberculosis infection, IL-22R, IL-17R and IFN-γ,R1 were highly expressed on the surface of PMC [ （86. 2±2. 9）%, （41.5± 4.4）% and （64. 9±5.8）% respectively]. （2） In group IL-22 ＋ IL-17, the percentage of Ki-67＋ PMC was（31.5± 2. 0）% and （26. 1± 2.4）% respectively, which were both higher than that in the mediumgroup [ （ 14. 6± 0. 7 ） % ] （ q = 6. 8 and 4. 9, respectively, both P 〈 0. 05 ）. In group IFN-γ, the percentage of Ki-67 ＋ PMC was（5.2 ±1.2） % , which was lower than that in the medium group （ q = 5.0, P 〈 0. Off）. In group IFN-γ ＋ IL-22 and IFN-γ ＋ IL-17, the percentages of Ki-67＋ PMC were （ 23.4±1.7 ） % and （21.8 ±3.8 ）% respectively, which were both higher than that in group IFN-γ （ q = 7.3 and 6.7, respectively, both P 〈 0. 05）. In group TPE and TPE ＋ IgG, the percentages of Ki-67 ＋ PMC were（63±9 ）% and（63 ±11 ）% respectively, which were both higher than that in the medium group （q = 19.6 and 19.7, respectively, both P 〈 0. 05）. In group TPE ＋ anti-IFN-γ mAb, the percentage of Ki-67＋PMC was （82± 4 ） % , which was even higher than that in group TPE ＋ IgG （ q = 7.5, P 〈 0. Off ）. In group TPE ＋ anti-IL-22 mAb, the percentage of Ki-67 ＋ PMC was （ 34±3 ） %, which was lower than that in group TPE ＋ IgG （q = 11.8, P 〈 0. 05 ）. In group TPE ＋ anti-IL-17 mAb, the percentage of Ki-67 ＋ PMC was （58±5 ） % , which showed no significant difference compared to that in group TPE ＋ IgG （ q = 2. 1, P 〉 O. 05 ）. （3） The percentage of apoptotic PMC in group IFN-γ was （19.3±1. 1 ）%, which was higher than that in the medium group [ （4. 3 ± 0. 6 ） %] （ q = 33.4, P 〈 0. 05 ）. The percentage of apoptotie PMC in group IL-22 ＋ IL-17 was （ 3.8± 0. 6 ） % and （ 5.7±0. 8 ） % respectively, which had no significant difference compared to that in the medium group （q = 1.3 and 3.0, respectively, both P 〉 0. 05 ）. The percentage of apoptotic PMC in group IFN-γ ＋ IL-22 and IFN-γ ＋ IL-17 were（6.5 ±0.7）% and（8.7±1.7）% respectively, which were both lower than that in group IFN-γ （ q = 28.5 and 23.6, respectively, both P 〈 0. 05 ）. Conclusion During Mycobacteriurn tuberculosis infection, IFN-γ inhibited PMC proliferation and contributed to apoptosis, while IL-22 and IL-17 promoted PMC proliferation without influencing PMC apoptosis and succeeded in reversing the effect induced by IFN-γ.