脑啡肽对湖北钉螺血淋巴细胞的影响
|
摘要
为观察脑啡肽(MENK)对湖北钉螺血淋巴细胞吞噬力的影响,将湖北钉螺软体组织挤压后获取钉螺血淋巴液,设9个实验组:钉螺血淋巴液对照组(A组)、 MENK溶剂组(B组)、钉螺血淋巴液+1 mg/L MENK组(C组)、钉螺血淋巴液+5 mg/L MENK组(D组)、钉螺血淋巴液+10 mg/L MENK组(E组)、钉螺血淋巴液+醛化绵羊红细胞组(F组)、钉螺血淋巴液+醛化绵羊红细胞+1 mg/L MENK组(G组)、钉螺血淋巴液+醛化绵羊红细胞+5 mg/L MENK组(H组)、钉螺血淋巴液+醛化绵羊红细胞+10 mg/L MENK组(I组)。将各组同时置25℃室温下孵育60 min后制成玻片涂片,经吖啶橙染色,于荧光显微镜下观察并统计各组不同形态类型的钉螺血淋巴细胞数。结果显示:钉螺血淋巴细胞中的伸展细胞可以直接吞噬异物(醛化绵羊红细胞); C、 G组(1 mg/L MENK组)伸展细胞的比率分别上升至48.77%(1 323/2 713)和48.30%(1 337/2 768),明显高于A组的39.99%(1 072/2 681),差异具有统计学意义P <0.01),而D、 H组(5 mg/L MENK组)和E、 I组(10 mg/L MENK组)的伸展细胞比率则分别下降至27.04%(706/2 611)、 26.29%(665/2 529)、 19.23%(422/2 194)和18.3%(414/2 262),与A组相比差异有统计学意义(P <0.01); G组(1 mg/L MENK组)对异物的吞噬率升高至48.77%(652/1 337),而H、 I两组对异物的吞噬率分别下降至7.37%(49/665)和5.8%(24/414),均与F组吞噬率40.09%(352/878)的差异具有统计学意义(P <0.01)。提示不同浓度MENK对钉螺血淋巴细胞的吞噬力有明显地促进或抑制作用。
To observe the effect of Met-enkephalin(MENK) on phagocytosis of hemocytes of Oncomelania hupensis, the hemolymph was obtained by squeezing the soft tissues of O. hupensis. The hemocytes in the hemolymph were incubated with different concentrations of MENK(0, 1, 5 and 10 mg/L) in the presence or absence of formaldehyde-treated sheep erythrocyte(FSE) at room temperature for 60 min. The hemocytes were stained with acridine orange and observed under fluorescence microscope. The results showed that the low concentration of MENK(1 mg/L) significantly stimulated the development of pseudopodia in the treated hemocytes of O. hupensis. The rates of pseudopodia development in the group treated with 1 mg/L of MENK was 48.77%(without FSE) and 48.3%(with FSE) with statistical significance compared to the control group(39.99%)( P < 0.01). The low concentration of MENK also stimulated the phagocytosis of treated hemocytes to FSE(48.77%) compared to control group without treatment( 40. 09 %) with significant difference( P < 0. 01). However, the high concentration of MENK( 5 mg/L or10 mg/L) not only inhibited hemocytes of Oncomelania to develop pseudopodia [27.04%(5 mg/L), 26.29%(5 mg/L +FSE), 19.23%(10 mg/L) and 18.3%(10 mg/L + FSE) with significant difference compared to control group( χ~2=99.366, 109.74, 244.446, 274.34, respectively, all P < 0.01), but also dramatically inhibited the phagocytosis rate of treated hemocytes to FSE [7.37%(5 mg/L) and 5.8%(10 mg/L) ] compared to 40.09% in untreated control group with statistical significance(P < 0.01). The results demonstrate that low concentrations of MENK can stimulate, but high concentration of MENK can inhibit the phagocytosis of treated hemocytes of O. hupensis.
To observe the effect of Met-enkephalin(MENK) on phagocytosis of hemocytes of Oncomelania hupensis, the hemolymph was obtained by squeezing the soft tissues of O. hupensis. The hemocytes in the hemolymph were incubated with different concentrations of MENK(0, 1, 5 and 10 mg/L) in the presence or absence of formaldehyde-treated sheep erythrocyte(FSE) at room temperature for 60 min. The hemocytes were stained with acridine orange and observed under fluorescence microscope. The results showed that the low concentration of MENK(1 mg/L) significantly stimulated the development of pseudopodia in the treated hemocytes of O. hupensis. The rates of pseudopodia development in the group treated with 1 mg/L of MENK was 48.77%(without FSE) and 48.3%(with FSE) with statistical significance compared to the control group(39.99%)( P < 0.01). The low concentration of MENK also stimulated the phagocytosis of treated hemocytes to FSE(48.77%) compared to control group without treatment( 40. 09 %) with significant difference( P < 0. 01). However, the high concentration of MENK( 5 mg/L or10 mg/L) not only inhibited hemocytes of Oncomelania to develop pseudopodia [27.04%(5 mg/L), 26.29%(5 mg/L +FSE), 19.23%(10 mg/L) and 18.3%(10 mg/L + FSE) with significant difference compared to control group( χ~2=99.366, 109.74, 244.446, 274.34, respectively, all P < 0.01), but also dramatically inhibited the phagocytosis rate of treated hemocytes to FSE [7.37%(5 mg/L) and 5.8%(10 mg/L) ] compared to 40.09% in untreated control group with statistical significance(P < 0.01). The results demonstrate that low concentrations of MENK can stimulate, but high concentration of MENK can inhibit the phagocytosis of treated hemocytes of O. hupensis.
引文
[1]Zagon IS,Rahn KA,Bonneau RH,et al.Opioid growth factor suppresses expression of experimental autoimmune encephalomyelitis[J].Brain Res,2010,15(7):154-161.
[2]张克莹,都健,单风平.蛋氨酸脑啡肽的免疫调节作用及应用新进展[J].中国免疫学杂志,2017,33(5):788-792.
[3]黄雯雯,王宜艳,管晓娟,等.柄袋沙蠋体壁和消化系统甲硫氨酸脑啡肽免疫组织化学定位[J].四川动物,2011,30(4):502-505.
[4]李华,孙虎山.贝类等无脊椎动物脑啡肽研究进展[J].动物学杂志,2012,47(6):128-135.
[5]李华,孙虎山,王宜艳,等.扁玉螺体表和消化系统甲硫氨酸脑啡肽免疫组织化学定位[J].动物学杂志,2008,43(4):72-76.
[6]Mancillas JR,McGinty JF,Selverston AI,et al.Immunocytochemical localization of enkephalin and substance P in retina and eyestalk neurones of lobster[J].Nature,1981,293(15):576-578.
[7]Stefano GB,Scharrer B.High affinity binding of an enkephalin analog in the cerebral ganglion of the insect Leucophaea maderae(Blattaria)[J].Brain Res,1981,225(1):107-114.
[8]张红梅,诸葛洪祥,王玉芳,等.钉螺血淋巴细胞及其功能的研究[J].中国寄生虫学与寄生虫病杂志,2007,25(2):141-145.
[9]谭苹,于志君.湖北钉螺中枢神经节的解剖方法[J].中国寄生虫学与寄生虫病杂志,2015,33(5):350-351.
[10]熊平源,王强.医学免疫学实验教程[M].武汉:武汉大学出版社,2011:42.
[11]卫向红,谭苹,万伦,等.吖啶橙染色法用于湖北钉螺血淋巴细胞的观察[J].中国寄生虫学与寄生虫病杂志,2018,36(3):315-317.
[12]王晓勤.腹足类血淋巴细胞的研究[J].国外医学寄生虫病分册,1994,21(2):52-55.
[13]王晓勤,裘丽姝,何毅勋,等.湖北钉螺血淋巴细胞的形态及其吞噬功能[J].中国寄生虫学与寄生虫病杂志,1994,12(2):100-103.
[14]Stefano GB,Leung MK,Zhao XH,et al.Evidence for the involvement of opioid neuropeptides in the adherence and migration of immunocompetent invertebrate hemocytes[J].Proc Natl Acad Sci USA,1989,86(2):626-630.
[15]魏凤华,覃杰,李博,等.氯代水杨胺缓释颗粒剂杀灭钉螺效果评价[J].中国寄生虫学与寄生虫病杂志,2016,34(4):308-314.
[16]杨蓬,段红颜,王策,等.紫皮独蒜液对日本血吸虫尾蚴和钉螺的杀灭效果[J].中国寄生虫学与寄生虫病杂志,2012,30(3):245-246.
[17]崔国艳,汪世平,程红兵,等.灭螺微生物的选育及其效果观察[J].中国寄生虫学与寄生虫病杂志,2015,33(3):234-236.
[18]谭苹,张学俊,杨建明,等.空心莲子草对钉螺酶组织化学的影响[J].中国寄生虫学与寄生虫病杂志,2009,27(1):11-16.
[2]张克莹,都健,单风平.蛋氨酸脑啡肽的免疫调节作用及应用新进展[J].中国免疫学杂志,2017,33(5):788-792.
[3]黄雯雯,王宜艳,管晓娟,等.柄袋沙蠋体壁和消化系统甲硫氨酸脑啡肽免疫组织化学定位[J].四川动物,2011,30(4):502-505.
[4]李华,孙虎山.贝类等无脊椎动物脑啡肽研究进展[J].动物学杂志,2012,47(6):128-135.
[5]李华,孙虎山,王宜艳,等.扁玉螺体表和消化系统甲硫氨酸脑啡肽免疫组织化学定位[J].动物学杂志,2008,43(4):72-76.
[6]Mancillas JR,McGinty JF,Selverston AI,et al.Immunocytochemical localization of enkephalin and substance P in retina and eyestalk neurones of lobster[J].Nature,1981,293(15):576-578.
[7]Stefano GB,Scharrer B.High affinity binding of an enkephalin analog in the cerebral ganglion of the insect Leucophaea maderae(Blattaria)[J].Brain Res,1981,225(1):107-114.
[8]张红梅,诸葛洪祥,王玉芳,等.钉螺血淋巴细胞及其功能的研究[J].中国寄生虫学与寄生虫病杂志,2007,25(2):141-145.
[9]谭苹,于志君.湖北钉螺中枢神经节的解剖方法[J].中国寄生虫学与寄生虫病杂志,2015,33(5):350-351.
[10]熊平源,王强.医学免疫学实验教程[M].武汉:武汉大学出版社,2011:42.
[11]卫向红,谭苹,万伦,等.吖啶橙染色法用于湖北钉螺血淋巴细胞的观察[J].中国寄生虫学与寄生虫病杂志,2018,36(3):315-317.
[12]王晓勤.腹足类血淋巴细胞的研究[J].国外医学寄生虫病分册,1994,21(2):52-55.
[13]王晓勤,裘丽姝,何毅勋,等.湖北钉螺血淋巴细胞的形态及其吞噬功能[J].中国寄生虫学与寄生虫病杂志,1994,12(2):100-103.
[14]Stefano GB,Leung MK,Zhao XH,et al.Evidence for the involvement of opioid neuropeptides in the adherence and migration of immunocompetent invertebrate hemocytes[J].Proc Natl Acad Sci USA,1989,86(2):626-630.
[15]魏凤华,覃杰,李博,等.氯代水杨胺缓释颗粒剂杀灭钉螺效果评价[J].中国寄生虫学与寄生虫病杂志,2016,34(4):308-314.
[16]杨蓬,段红颜,王策,等.紫皮独蒜液对日本血吸虫尾蚴和钉螺的杀灭效果[J].中国寄生虫学与寄生虫病杂志,2012,30(3):245-246.
[17]崔国艳,汪世平,程红兵,等.灭螺微生物的选育及其效果观察[J].中国寄生虫学与寄生虫病杂志,2015,33(3):234-236.
[18]谭苹,张学俊,杨建明,等.空心莲子草对钉螺酶组织化学的影响[J].中国寄生虫学与寄生虫病杂志,2009,27(1):11-16.