滋阴降胃法对糖尿病胃轻瘫大鼠十二指肠Cajal间质细胞ICC和肌间神经丛的影响

Effect of Ziyin Jiangwei method on interstitial cells of Cajal and myenteric nerve plexus in a diabetic gastroparesis rat model

目的探讨滋阴降胃法对糖尿病胃轻瘫(diabetic gastroparesis,DGP)大鼠十二指肠Cajal间质细胞(interstitial cells of Cajal,ICC)和肌间神经丛的影响。方法鼠龄8周的SPF级雄性SD大鼠61只,按随机数字表法分为正常对照组10只及造模组51只。造模组采用链脲佐菌素(streptozotocin,STZ)法,将成模大鼠按随机数字表法分为模型组11只、多潘立酮组及滋阴降胃法中药高、中、低剂量组各10只,多潘立酮组予多潘立酮2.92 mg/(kg·d)灌胃,滋阴降胃法中药高、中、低剂量组分别按照含生药量12.5 g/(kg·d)、6.25 g/(kg·d)和3.125 g/(kg·d)灌胃,模型组和正常对照组予同等容量无菌水灌胃,连续干预8周。分别于干预0、4、8周采血测定血糖;干预8周后麻醉处死全部大鼠,测定小肠传输速率,免疫组化法检测十二指肠c-kit、突触素(synaptophysin,SYN)、蛋白基因产物9.5(protein gene product 9.5,PGP9.5)的表达水平。实验过程中模型组和滋阴降胃法中剂量组大鼠各死亡1只。结果与正常对照组比较,各时间点模型组体重均下降,血糖均明显升高,模型组小肠传输速率、十二指肠c-kit、SYN和PGP9.5表达明显下降,差异均具有统计学意义(P<0.05)。与模型组比较,8周时滋阴降胃法各剂量组体重均明显升高,4周时滋阴降胃法高剂量组血糖明显下降,滋阴降胃法中各剂量组小肠传输速率明显升高,滋阴降胃法中各剂量组十二指肠c-kit、SYN和PGP9.5表达明显升高,差异均具有统计学意义(P<0.05)。结论滋阴降胃法中药可能通过调节ICC、肌间神经丛来增加胃肠动力,从而治疗DGP。

Objective To explore the effects of Ziyin Jiangwei method on interstitial cells of Cajal(ICC)and myenteric nerve plexus in diabetic gastroparesis(DGP)rats.Methods Sixty-one Sprague Dawley male rats(eight-week old in specific pathogen free)were divided into the normal control group(n=10)and modeling-ready group(n=51)by a random number table.Streptozotocin was used in model establishment.The modeling rats were further assigned to the model group(n=11),domperidone group(n=10),and Ziyin Jiangwei method groups(high,medium and low,n=10 each group)by a random number table.In the domperidone group,2.92 mg/(kg·d)domperidone was used for gavage.In the Ziyin Jiangwei method groups,they were provided with gavage at the dosages of 12.5 g/(kg·d),6.25 g/(kg·d),and 3.125 g/(kg·d)at high,medium,and low levels respectively.Both the model group and normal control group were provided with the same volume of sterile drinking water for gavage.The intervention lasted for eight weeks.On Weeks 0,4,and 8,the blood glucose were measured.After eight-week intervention,all rats were sacrificed after anesthesia.The small intestine transmission rate was tested.The levels of duodenal c-kit,synaptophysin(SYN)and protein gene product 9.5(PGP9.5)were measured by immunohistochemistry.During experiment,one death occurred in each model group and medium dose Ziyin Jiangwei method group.Results As compared with the normal control group,the levels of body weight at different time points in the model group all decreased and the levels of blood glucose all increased,and the small intestine transmission rate and the levels of duodenal c-kit,SYN and PGP9.5 in the model group all significantly decreased with statistical significance(P<0.05).As compared with the model group,at 8 weeks,the body weight of each dose group increased significantly.At 4 weeks,blood glucose in the high-dose group of Ziyin Jiangwei prescription decreased significantly.The small intestine transmission rate,and c-kit,SYN and PGP9.5 all increased in high-,medium-,and low-dose Ziyin Jiangwei method groups with statistical significance(P<0.05).Conclusions Ziyin Jiangwei method in the treatment of DGP is likely to be realized by regulating ICC and myenteric nerve plexus to enhance gastrointestinal motility.