益气养阴化瘀通络中药对糖尿病胃轻瘫大鼠平滑肌细胞凋亡的影响

Effect of Yiqi Yangyin Huayu Tongluo Recipe on apoptosis of gastric smooth muscle cells in rats with diabetic gastroparesis

目的:研究益气养阴化瘀通络中药对糖尿病胃轻瘫大鼠中氧化应激和细胞凋亡的影响.方法:30只♂SD大鼠随机分为正常对照组(normal control group,NC组)、糖尿病模型组(diabetic model group,DM组)、中药干预组(Chinese medicine group,ZY组).予腹腔注射链脲佐菌素(streptozotocin,STZ)制备糖尿病大鼠模型,中药干预组予以中药灌胃,其余两组分别予等体积生理盐水灌胃.6 wk后检测胃排空率;测定胃组织超氧化物歧化酶(superoxide dismutase,SOD)水平及丙二醛(malondialdehyde,MDA)含量;原位末端标记(TUNEL)法检测胃平滑肌细胞凋亡;Real-time PCR法检测胃组织Bcl-2及Bax mRNA表达.结果:与NC组相比,DM组及ZY组大鼠胃排空率明显下降(62.64%±4.51%vs 18.65%±4.30%,26.84%±2.35%,P<0.05),与DM组相比,ZY组胃排空率明显升高(18.65%±4.30%vs 26.84%±2.35%,P<0.05);与NC组相比,DM组及ZY组大鼠胃组织MDA含量均明显升高(2.12 nmol/mgprot±0.08 nmol/mgprot vs 7.55nmol/mgprot±0.27 nmol/mgprot,2.82 nmol/mgprot±0.16 nmol/mgprot,P<0.05),ZY组MDA含量较DM组明显降低(2.82 nmol/mgprot±0.16 nmol/mgprot vs 7.55 nmol/mgprot±0.27nmol/mgprot,P<0.05);DM组及ZY组大鼠胃组织SOD活性较NC组均明显降低(48.18 U/mgprot±1.46 U/mgprot,73.66 U/mgprot±0.55U/mgprot vs 120.20 U/mgprot±1.21 U/mgprot,P<0.05),与DM组相比,ZY组大鼠胃组织SOD活性较DM组明显升高(48.18 U/mgprot±1.46U/mgprot vs 73.66 U/mgprot±0.55 U/mgprot,P<0.05);DM组及ZY组胃平滑肌凋亡细胞数目较NC组明显增多(10.84%±1.18%,1.73%±0.09%vs 0.28%±0.25%,P<0.05),ZY组大鼠胃平滑肌细胞凋亡数量明显较DM组明显减少(1.73%±0.09%vs 10.84%±1.18%,P<0.05);DM组和ZY组Bcl-2 mRNA表达水平较NC组明显降低(1.65±0.28 vs 0.59±0.09,1.10±0.27,P<0.05),ZY组Bcl-2 mRNA表达水平较DM组明显升高,差异具有统计学意义(1.10±0.27 vs 0.59±0.09,P<0.05);与NC组相比,DM组Bax mRNA表达水平显著升高(0.87±0.83vs 1.96±0.28,P<0.05),ZY组Bax mRNA表达水平没有明显变化((0.87±0.83 vs 1.13±0.11,P>0.05);ZY组Bax mRNA表达水平明显低于DM组(1.13±0.11 vs 1.96±0.28,P<0.05).结论:益气养阴化瘀通络中药可通过提高抗氧化能力,抑制氧化应激及细胞凋亡而促进胃动力.

AIM： To explore the effect of Yiqi Yangyin Huayu Tongluo Recipe on oxidative stress and apoptosis of gastric smooth muscle cells in rats with diabetic gastroparesis. METHODS： Thirty Sprague-Dawley male rats were randomly divided into three groups： a normal control （NC） group, a diabetic model （DM） group and a Chinese medicine （ZY）-group. Diabetes was induced by injecting streptozotocin（STZ）. The Chinese medicine group was treated with traditional Chinese medicine, while the other two groups received the same volume of saline. Six weeks later, rate of gastric emptying was measured with phenol red; levels of malondialdehyde （MDA） and activity of superoxide dismutase （SOD） in gastric tissues were detected; apoptosis of gastric smooth muscle cells was detected by TUNEL assay; and expression of Bcl-2 and Bax mRNAs was examined by realtime PCR. RESULTS： Compared with the NC group, the rates of gastric emptying in the DM and ZY groups decreased significantly （62.64% ± 4.51% vs 18.65% ± 4.30%, 26.84% ± 2.35%, P 〈 0.05 for both）, while the gastric emptying rate was significantly higher in the ZY group than in the DM group （P 〈 0.05）. Compared with the NC group, gastric MDA contents were significantly elevated in the DM and ZY groups （2.12 nmol/mgprot ± 0.08 nmol/mgprot vs 7.55 nmol/mgprot ± 0.27 nmol/mgprot, 2.82 nmol/mgprot ± 0.16 nmol/ mgprot, P 〈 0.05 for both）, while gastric MDA content was significantly lower in the ZY group than in the DM group （P 〈 0.05）. SOD activity was reduced significantly in the DM and ZY groups in comparison with the NC group （48.18 U/mgprot ± 1.46 U/mgprot, 73.66 U/mgprot ± 0.55 U/mgprot vs 120.20 U/mgprot ± 1.21 U/ mgprot, P 〈 0.05 for both）, while SOD activity was significantly higher in the ZY group than in the DM group （P 〈 0.05）. The apoptotic index （AI） of gastric smooth muscle cells was significantly increased in the DM and ZY groups compared with the NC group （10.84% ± 1.18%, 1.73% ± 0.09% vs 0.28% ± 0.25%, P 〈 0.05 for both）, while the AI was significantly lower in the ZY group than in the DM group （P 〈 0.05）. The levels of Bcl-2 mRNA in the DM and ZY groups were significantly lower compared with the NC group （1.65 ± 0.28 vs 0.59 ± 0.09, 1.10 ± 0.27, P 〈 0.05 for both）, while the level of Bcl-2 mRNA was significantly higher in the ZY group than in the DM group （P 〈 0.05）. The expression of Bax mRNA in the DM group was significantly higher than that in the NC group （1.96 ± 0.28 vs 0.87 ± 0.83, P 〈 0.05）, while there was no significant difference between the NC group and ZY group （0.87 ± 0.83 vs 1.13 ± 0.11, P 〉 0.05）. Compared with the DM group, the expression of Bax mRNA in the ZY group was significantly decreased （1.96 ± 0.28 vs 1.13 ± 0.11, P 〈 0.05）. CONCLUSION： Yiqi Yangyin Huayu Tongluo Recipe can improve gastric motility by increasing antioxidative action, decreasing oxidative stress and inhibiting apoptosis in rats with diabetic gastroparesis.