黑枸杞水提物对中波紫外线辐射后人角质形成细胞增殖与凋亡及凋亡相关蛋白表达水平的影响研究

Effects of Black Wolfberry Water Extracts on Proliferation and Apoptosis and Apoptosis-related-protein Expression Level of HaCaT Cells after UVB Radiation

目的研究黑枸杞水提物对中波紫外线(UVB)辐射后人角质形成(HaCaT)细胞增殖与凋亡及凋亡相关蛋白[P38丝裂原活化蛋白激酶(P38MAPK)、P53、半胱氨酸蛋白酶-8(caspase-8)、半胱氨酸蛋白酶-3(caspase-3)、B细胞淋巴瘤因子2(Bcl-2)]表达水平的影响,以期探讨黑枸杞水提物减轻UVB损伤HaCaT细胞的作用机制。方法 2015年3月—2016年1月,提取黑枸杞水提物,体外培养HaCaT细胞,取对数生长期的HaCaT细胞,采用随机数字表法分为对照组(无辐射)、UVB组(30 m J/cm^2UVB辐射40 min)、黑枸杞水提物组(30 mJ/cm^2UVB辐射40 min+黑枸杞水提物2 mg/ml)。采用MTS法检测各组细胞增殖活力,流式细胞仪检测各组细胞凋亡率,Western blotting法检测各组P38MAPK、P53、caspase-8、caspase-3、Bcl-2表达水平。结果 UVB组细胞增殖活力小于对照组(P<0.05);黑枸杞水提物组细胞增殖活力小于对照组,大于UVB组(P<0.05)。UVB组细胞凋亡率大于对照组(P<0.05);黑枸杞水提物组细胞凋亡率大于对照组,小于UVB组(P<0.05)。UVB组P38MAPK、P53、caspase-8、caspase-3表达水平高于对照组,Bcl-2表达水平低于对照组(P<0.05);黑枸杞水提物组P38MAPK、P53、caspase-8、caspase-3表达水平低于UVB组,Bcl-2表达水平高于UVB组(P<0.05)。结论黑枸杞水提物可促进UVB辐射后HaCaT细胞的增殖,同时阻止其凋亡,其机制可能为黑枸杞水提物能够下调P38MAPK、P53、caspase-8、caspase-3表达水平及上调Bcl-2表达水平,进而减轻HaCaT细胞的辐射损伤。

Objective To research the effect of black wolfberry water extracts on proliferation and apoptosis and P38 mitogen activated protein kinase （ P38MAPK）, P53, cysteine proteinase 8 （ caspase - 8） , eysteine proteinase 3 （ caspase - 3 ） and B cell lymphoma factor 2 （ Bcl - 2） expression of HaCaT cells after ultraviolet B （UVB） radiation, in order to explore the mechanism of black wolfberry water extracts reducing the damage of HaCaT cells after UVB radiation. Methods From March 2015 to January 2016, black wolfberry water extracts was extracted and HaCaT cells were cultured in vitro. HaCaT cells in logarithmic growth phase were collected and randomly divided into 3 groups by random number table method： control group （without radiation）, UVB group （30 mJ/cm2 UVB radiation for 40 rain） and black wolfberry water extracts group （30 mJ/cm2 UVB radiation for 40 rain ＋ black wolfberry water extracts 2 rag/m1） . Cell proliferation in each group was detected by MTS, apoptotic rate in each group was assessed by flow cytometry. The expression levels of P38MAPK, P53, caspase -8, caspase -3 and Bcl -2 in each group were detected by Western blotting. Results Cell proliferation in UVB group was lower than that in control group （P 〈 0.05 ） ; cell proliferation in black wolfberry water extracts group was lower than that in control group but higher than that in UVB group （P 〈 0. 05 ） . Apoptotic rate in UVB group was higher than that in control group （P 〈 0. 05 ） ; apoptotic rate in black wolfberry water extracts group was higher than that it/control group but lower than that in UVB group （ P 〈0. 05） . The expression levels of P38MAPK, P53, caspase - 8, caspase - 3 in UVB group were higher than those in control group, but the expression level of Bcl - 2 in UVB group was lower than that in control group （ P 〈 0. 05 ） ; the expression levels of P38MAPK, P53, caspase- 8, caspase -3 in black wolfberry water extracts group were lower than those in UVB group, but the expression level of Bcl - 2 in black wolfberry water extracts group was higher than that in UVB group （ P 〈 0. 05 ） . Conclusion The black wolfberry water extracts can promote the proliferation of HaCaT cells after UVB radiation and prevent their apoptosis. The mechanism might be that the black wolfberry water extracts can down -regulate the expression levels of P38 MAPK, P53, caspase - 8 and caspase - 3 and up - regulate the expression level of Bcl - 2, so as to reduce the radiation damage of HaCaT cells.