摘要
目的观察不同浓度的钙蛋白酶抑制剂ALLN对C2C12成肌细胞增殖和凋亡的影响。方法Ca^2＋和ALLN干预C2C12细胞后，采用噻唑蓝法和流式细胞术检测细胞增殖和凋亡情况，采用Giemsa染色观察细胞形态。结果0．5、1、2、4、8、16、32、64、128mmol／L Ca^2＋干预C2C12细胞72h的吸光度（A）值显著低于对照组（均P〈0．05）；16mmol／L的Ca^2＋和AUN干预6、12、24、36h后，AUN1—7组（含16mmol／LCa^2＋和AIJN终浓度为3．125、6．25、12．5、25、50、100、200 μmol／L的无血清培养基）A值均显著高于Ca^2＋组（干预6h：0．449±0．024、0．472±0．022、0．513±0．008、0．540±0．014、0．588±0．016、0．607±0．030、0．700±0．020比0．355±0．012，P值均为0．000；干预12h：0．407±0．007、0．414±0．006、0．434±0．004、0．441±0．003、0．460±0．010、0．484±0．006、0．525±0．006比0．368±0．027，P值均为0．000；干预24h：0．436±0．005、0．431±0．015、0．441±0．006、0．459±0．013、0．527±0．009、0．581±0．005、0．599±0．011比0．386±0．007，P值均为0．000；干预36h：0．464±0．022、0．460±0．018、0．461±0．007、0．434±0．020、0．454±0．028、0．479±0．006、0．524±0．011比0．379±0．011，P值均为0．000），干预48～72h后差异无统计学意义。36h时ALLN10、50、100、200μmol／L组的凋亡率分别为（6．00±1．20）％、（5．02±1．13）％、（4．89±1．11）％、（2．71±1．15）％，均显著低于Ca^2＋组（13．70±2．30）％（P值均为0．000）。Giemsa染色显示Ca^2＋组出现细胞凋亡形态学改变，ALLN组细胞凋亡情况明显改善。结论16mmol／L的Ca^2＋可诱导C2C12细胞凋亡，ALLN可抑制细胞凋亡、促进增殖，该作用呈时间和剂量依赖性。
Objective To explore the effect of different concentrations of ALLN on proliferation and apoptosis of C2C12 myoblasts. Methods After intervention with Ca^2＋ and ALLN, methyl thiazolyl tetrazolium and flow cytometry were used to determine the effect of Ca^2＋ and ALLN on the proliferation and apoptosis of C2C12 cells, respectively. The morphological changes of C2C12 myoblasts were observed using Giemsa staining. Results The absorbance of Ca^2＋ group was significantly lower than that of the control group （ P 〈 0. 05）. After 6, 12, 24, 36 hours of intervention, the absorbance in ALLN groups 1 to 7 （ cultured in serum-free media containing 16 mmol/L Ca^2＋ and ALLN at final concentrations of 3. 125, 6. 25, 12. 5, 25, 50, 100, 200 μmol/L） were all significantly higher than that in the 16 mmol/L Ca^2＋ group （after 6 hours： 0. 449 ±0. 024, 0. 472 ±0. 022, 0. 513 ±0. 008, 0. 540 ±0. 014, 0. 588 ±0. 016, 0. 607 ±0. 030, 0. 700 ±0. 020 vs. 0. 355 ±0. 012, all P =0. 000; after 12 hours： 0. 407 ±0. 007, 0. 414 ±0. 006, 0. 434 ±0. 004, 0. 441 ± 0.003, 0.460±0.010, 0.484 ±0.006, 0.525 ±0.006 vs. 0.368 ±0.027, allP=0.000; after 24 hours： 0.436 ±0.005, 0.431 ±0.015, 0.441 ±0.006, 0.459 ±0.013, 0.527 ±0.009, 0.581 ±0.005, 0.599 ±0. 011 vs. 0. 386 ±0. 007, all P =0. 000; after 36 hours： 0. 464 ±0. 022, 0. 460 ±0. 018,0. 461 ±0. 007, 0.434±0.020, 0.454 ±0.028, 0.479 ±0.006, 0.524 ±0.011 vs. 0.379 ±0.011, all P=0.000）, while no significant differences were observed after 48 -72 hours of intervention. After treatment for 36 hours, the apoptosis rate in ALLN 10, 50, 100, and 200 μmol/L groups were （6. 00 ±1.20）%, （5.02 ±1.13 ）%, （4. 89 ±1.11 ）%, and （2.71 ±1.15 ）%, all significantly lower than that in the Ca^2＋ group [ （13.70 ±2. 30） % ] （ all P = 0. 000）. Giemsa staining showed apoptotic morphological changes in the Ca^2＋ group, which were obviously alleviated in the ALLN group. Conclusions Ca^2＋ at a concentration of 16mmol/L can induce apoptosis of C2C12 ceils. In contrast, ALLN can inhibit cell apoptosis and promote proliferation in a time- and dose-dependent manner.
出处
《中华临床营养杂志》
CAS
CSCD
2015年第1期35-40,共6页
Chinese Journal of Clinical Nutrition
基金
国家自然科学基金（81272465）
