女贞子多糖在大鼠睾丸支持细胞炎性损伤中对能量代谢保护作用的研究
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摘要
目的探讨女贞子多糖(PLL)对脂多糖(LPS)诱导的睾丸支持细胞炎性损伤模型大鼠能量代谢的影响。方法体外分离和培养大鼠支持细胞,分对照组、LPS组、LPS+低剂量PLL组、LPS+中剂量PLL组和LPS+高剂量PLL组。CCK-8检测细胞增殖;紫外分光光度法检测细胞培养液上清中三磷酸腺苷(ATP)、乳酸(LD)、乳酸脱氢酶(LDH)和琥珀酸脱氢酶(SDH)活力。结果对照组、LPS组、LPS+低剂量PLL组、LPS+中剂量PLL组和LPS+高剂量PLL组细胞增殖检测OD值分别为:(0.65±0.02)%、(0.32±0.02)%、(0.38±0.02)%、(0.45±0.02)%和(0.53±0.02)%,组间差异有统计学意义(F=152.30,P<0.01),LPS组与对照组相比细胞增殖率明显降低(P<0.01),而中剂量和高剂量LPS+PLL组与LPS组相比细胞增殖率增高(P<0.01);对照组、LPS组、LPS+低剂量PLL组、LPS+中剂量PLL组和LPS+高剂量PLL组细胞ATP含量分别为:(87.47±4.88),(36.41±3.67),(40.34±4.40),(55.35±6.54),(65.34±5.27)nmol/mL。LD含量分别为:(107.21±7.47),(44.91±3.39),(49.54±4.67),(55.32±5.44),(76.08±0.42)nmol/mL,组间差异均有统计学意义(P均<0.01);LPS组与对照组相比ATP和LD含量均明显降低(P均<0.01)。中剂量和高剂量PLL+LPS组与LPS组相比ATP和LD含量均有不同程度升高(P<0.05或<0.01);对照组、LPS组、LPS+低剂量PLL组、LPS+中剂量PLL组和LPS+高剂量PLL组细胞LDH活力分别为:(98.54±6.59),(34.25±3.25),(39.27±4.24),(45.09±7.54),(57.54±4.37)U/mL。SDH活力分别为:(132.54±7.44),(54.24±4.24),(59.87±5.34),(65.62±7.27),(72.13±6.37)U/mL,组间差异均有统计学意义(P均<0.01),LPS组与对照组相比LDH、SDH活力均明显降低(P均<0.01)。中剂量和高剂量PLL+LPS组与LPS组相比LDH、SDH活力均有不同程度升高(P<0.05或<0.01)。结论女贞子多糖在LPS诱导睾丸支持细胞炎症反应过程中对细胞代谢具有保护作用。
Objective To explore the effect of polysaccharide extracted from glossy privet fruit(Ligustrum lucidum)(PLL)on energy metabolism in rats with inflammatory injury of testicular Sertoli cells induced by lipopolysaccharide(LPS). Methods Rat Sertoli cells were isolated and cultured in vitro, and cells were divided into control group, LPS group, LPS +low dose PLL group, LPS +medium dose PLL group and LPS +high dose PLL group. Cell proliferation was detected by CCK-8; Activities of adenosine triphosphate(ATP), lactic acid(LD), lactate dehydrogenase(LDH) and succinate dehydrogenase(SDH) in supernatant of cell culture medium were measured by ultraviolet spectrophotometry. Results The OD values of cell proliferation in control group, LPS group, LPS+low dose PLL group, LPS+medium dose PLL group and LPS+high dose PLL group were as the following:(0.65±0.02)%,(0.32±0.02)%,(0.38±0.02)%,(0.45±0.02)% and(0.53±0.02)%, respectively. There was significant difference between the groups(F=153.93, P<0.01). Compared to the control group, cell proliferation rate of LPS group reduced(P<0.01), and those of LPS+middle dose LLP group and LPS+high dose LLP group raised, compared with LPS group(P <0.01); ATP concentration of the above groups was(87.47 ±4.88),(36.41 ±3.67),(40.34 ±4.40),(55.35 ±6.54) and(65.34 ±5.27)nmol/mL, respectively. LD concentration was(107.21 ±7.47),(44.91 ±3.39),(49.54 ±4.67),(55.32±5.44) and(76.08±0.42)nmol/mL, respectively, and there was significant difference between the groups(P<0.01). Compared to the control group, both ATP and LD concentration decreased(P<0.01). But both middle and high dose group's ATP and LD concentration increased(P <0.05 and P <0.01). LDH activity was(98.54 ±6.59),(34.25±3.25),(39.27±4.24),(45.09±7.54) and(57.54±4.37)U/mL, respectively. SDH activity was(132.54±7.44),(54.24±4.24),(59.87±5.34),(65.62±7.27) and(72.13±6.37)U/mL, respectively, and there was significant difference between the groups(P<0.01). Both LDH and SDH concentration in LPS group decreased, compared with control group(P<0.01). But both middle and high dose group's LDH and SDH concentration increased, compared with LPS group(P<0.05 and P<0.01). Conclusion Ligustrum lucidum polysaccharide has a protective effect on cell metabolism in LPS-induced inflammatory response of testicular Sertoli cells.
Objective To explore the effect of polysaccharide extracted from glossy privet fruit(Ligustrum lucidum)(PLL)on energy metabolism in rats with inflammatory injury of testicular Sertoli cells induced by lipopolysaccharide(LPS). Methods Rat Sertoli cells were isolated and cultured in vitro, and cells were divided into control group, LPS group, LPS +low dose PLL group, LPS +medium dose PLL group and LPS +high dose PLL group. Cell proliferation was detected by CCK-8; Activities of adenosine triphosphate(ATP), lactic acid(LD), lactate dehydrogenase(LDH) and succinate dehydrogenase(SDH) in supernatant of cell culture medium were measured by ultraviolet spectrophotometry. Results The OD values of cell proliferation in control group, LPS group, LPS+low dose PLL group, LPS+medium dose PLL group and LPS+high dose PLL group were as the following:(0.65±0.02)%,(0.32±0.02)%,(0.38±0.02)%,(0.45±0.02)% and(0.53±0.02)%, respectively. There was significant difference between the groups(F=153.93, P<0.01). Compared to the control group, cell proliferation rate of LPS group reduced(P<0.01), and those of LPS+middle dose LLP group and LPS+high dose LLP group raised, compared with LPS group(P <0.01); ATP concentration of the above groups was(87.47 ±4.88),(36.41 ±3.67),(40.34 ±4.40),(55.35 ±6.54) and(65.34 ±5.27)nmol/mL, respectively. LD concentration was(107.21 ±7.47),(44.91 ±3.39),(49.54 ±4.67),(55.32±5.44) and(76.08±0.42)nmol/mL, respectively, and there was significant difference between the groups(P<0.01). Compared to the control group, both ATP and LD concentration decreased(P<0.01). But both middle and high dose group's ATP and LD concentration increased(P <0.05 and P <0.01). LDH activity was(98.54 ±6.59),(34.25±3.25),(39.27±4.24),(45.09±7.54) and(57.54±4.37)U/mL, respectively. SDH activity was(132.54±7.44),(54.24±4.24),(59.87±5.34),(65.62±7.27) and(72.13±6.37)U/mL, respectively, and there was significant difference between the groups(P<0.01). Both LDH and SDH concentration in LPS group decreased, compared with control group(P<0.01). But both middle and high dose group's LDH and SDH concentration increased, compared with LPS group(P<0.05 and P<0.01). Conclusion Ligustrum lucidum polysaccharide has a protective effect on cell metabolism in LPS-induced inflammatory response of testicular Sertoli cells.
引文
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[8]Boussouar F,Benahmed M.Lactate and energy metabolism in male germ cells[J].Trends Endocrinol Metab,2004,15(3):345-350.
[9]Hamdy AA,Aly,David A,et al.Bacterial lipopolysaccharideinduced oxidative stress in adult rat Sertoli cells in vitro[J].Toxicology in Vitro,2010,24(6):1266-1272.
[10]魏祥燕,王国娟,王桦影,等.女贞子药理作用研究进展[J].上海中医药杂志,2017,51(8):106-108.
[2] Riccioli A,Starace D,Galli R,et al.Sertoli cells initiate testicular innate immune responses through TLR activation[J].Journal of Immunology,2006,177(10):7122-7130.
[3]刘亭亭,王萌.女贞子化学成分与药理作用研究进展[J].中国实验方剂学杂志,2014,20(14):228-34.
[4]麦海星,陈立军,曲楠,等.一种获得高纯度睾丸支持细胞的分离方法[J].中国组织工程研究与临床康复,2010,14(31):5797-5800.
[5] Lee NP,Cheng CY.Adaptors,junction dynamics,and spermatogenesis[J].Biol Reprod,2004,71(21):392-404.
[6] Alves MG,Rato L,Carvalho RA,et al.Hormonal control ofSertoll cell metabolism regulates spermatogenesis[J].Cell Mol Life Sci,2013,70(5):777-793.
[7]Robinson R,Fritz I. Metabolism of glucose by Sertoli cells in culture[J].Biol Reprod,1981,24(2):1032-1041.
[8]Boussouar F,Benahmed M.Lactate and energy metabolism in male germ cells[J].Trends Endocrinol Metab,2004,15(3):345-350.
[9]Hamdy AA,Aly,David A,et al.Bacterial lipopolysaccharideinduced oxidative stress in adult rat Sertoli cells in vitro[J].Toxicology in Vitro,2010,24(6):1266-1272.
[10]魏祥燕,王国娟,王桦影,等.女贞子药理作用研究进展[J].上海中医药杂志,2017,51(8):106-108.