钩藤总生物碱对腹腔粘连大鼠内皮功能、炎性介质和粘连组织相关因子表达的影响

Effect of total uncaria alkaloid on the endothelial function of peritoneal adhesion rats and on the expression of related tissue factors

目的观察钩藤总生物碱对大鼠腹腔粘连形成、炎性介质和腹腔粘连组织羟脯氨酸(OHP)、可溶性细胞间黏附分子-1(sICAM-1)及转化生长因子-β_1(TGF-β_1)表达的影响。方法按随机数字表法将60只清洁级Wistar大鼠分为6组:假手术组、模型组、钩藤总生物碱低、中、高剂量组及对照组。假手术组仅开腹,其余组大鼠开腹并建立腹腔粘连模型。造模后,钩藤总生物碱各剂量组腹腔注射钩藤总生物碱(低剂量:25 mg/kg,中剂量:50 mg/kg,高剂量:100 mg/kg),对照组给予腹腔注射10 mg/kg地塞米松磷酸钠注射液,假手术组与模型组给予腹腔注射同体积的生理盐水,均为一日1次。各组均持续治疗4周。观察大鼠腹腔粘连程度,并对比纤维蛋白原(FIB)、纤维蛋白降解产物(FDP)、OHP、sICAM-1、TGF-β_1及其他相关指标的变化。结果 (1)腹腔粘连率和粘连程度分级:与假手术组对比,模型组的腹腔粘连等级评分升高(P<0.01);与模型组对比,钩藤总生物碱各剂量组的腹腔粘连等级评分均降低(P<0.05或P<0.01);(2)血象:与假手术组对比,模型组的白细胞计数(WBC)、FIB、OHP均明显升高,FDP降低(P<0.01);与模型组对比,钩藤总生物碱各剂量组的WBC、FIB、OHP均有所降低,FDP升高(P<0.05或P<0.01);(3)腹腔粘连组织sICAM-1、TGF-β_1:与假手术组对比,模型组的sICAM-1、TGF-β_1均明显升高(P<0.01);与模型组对比,钩藤总生物碱各剂量组的sICAM-1、TGF-β_1均有所降低(P<0.05或P<0.01);(4)炎性介质:与假手术组对比,模型组的血清超氧化物歧化酶(SOD)降低,丙二醛(MDA)、肿瘤坏死因子-α(TNF-α)、一氧化氮(NO)、白介素(IL)-1β、基质金属蛋白酶抑制物-1(TIMP-1)、基质金属蛋白-9(MMP-9)均明显升高(P<0.01);与模型组对比,钩藤总生物碱各剂量组的血清SOD升高,MDA、TNF-α、NO、IL-1β、TIMP-1、MMP-9均有所降低(P<0.05或P<0.01);(5)内皮功能:与假手术组对比,模型组的血清血管内皮生长因子(VEGF)、CD34均明显升高(P<0.01);与模型组对比,钩藤总生物碱各剂量组的血清VEGF、CD34均有所降低(P<0.05或P<0.01)。结论钩藤总生物碱能够改善腹腔粘连大鼠的腹腔粘连程度,可能与调控FIB、sICAM-1、TGF-β_1的水平,改善内皮功能及抑制炎性反应密切相关。

Objective To observe the effect of total uncaria alkaloid on the formation of peritoneal adhesion and on the expression of OHP, sICAM-1 and TGF-β_1 in peritoneal adhesion rats. Methods A total of 60 clean Wistar rats were divided into 6 groups: sham operation group, model group, total uncaria alkaloids with low, medium and high dose group and control group. The rats of sham operation group were only sliced abdomen, the rats in other 5 groups were sliced abdomen, and set up peritoneal adhesion model. After model building, the total uncaria alkaloid groups were given injection of total uncaria alkaloid(low dose: 25 mg/kg, medium dose: 50 mg/kg, high doses: 100 mg/kg) daily, control group were given 10 mg/kg intraperitoneal injection of dexamethasone sodium phosphate, sham operation group and model group were given intraperitoneal injection of volume of normal saline. After 4 week treatment, the peritoneal adhesion grade in rats was observed, and the contrast of FIB, FDP, OHP, sICAM-1, the TGF-β_1 change and other indexes were observed. Results(1)Peritoneal adhesion rate and grade of adhesion degree: compared with sham operation group, grade of peritoneal adhesion degree degree in model group was higher(P<0.01). Compared with model group, the grade of peritoneal adhesion were decreased in all doses of total uncaria alkaloid groups(P<0.05 or P<0.01);(2)Hemogram: compared with sham operation group, the WBC, FIB and OHP in model group were significantly increased, the FDP was decreased(P<0.01). Compared with model group, the WBC, FIB and OHP in all doses of total uncaria alkaloid groups were decreased, but the FDP was increased(P<0.05 or P<0.01);(3)Peritoneal adhesion tissue sICAM-1, TGF-β_1: compared with sham operation group, the sICAM-1 and TGF-β_1 in model group were significantly increased(P<0.01). Compared with model group, the sICAM-1 and TGF-β_1 in all doses of total uncaria alkaloid groups were decreased(P<0.05 or P<0.01).(4)Inflammatory medium: compared with sham operation group, the SOD in model group was significantly decreased, the MDA, TNF-α, NO, IL-1β, TIMP-1 and MMP-9 were significantly increased(P<0.01). Compared with model group, SOD in all doses of total uncaria alkaloid groups were significantly increased, while the MDA, TNF-α, NO, IL-1β, TIMP-1 and MMP-9 were decreased(P<0.05 or P<0.01).(5)Endothelial function: compared with the sham operation group, the serum VEGF and CD34 of the model group were significantly increased(P<0.01). Compared with the model group, the serum VEGF and CD34 of all doses of total uncaria alkaloid groups were decreased(P<0.05 or P<0.01). Conclusion Total uncaria alkaloid can improve the peritoneal adhesion degree in peritoneal adhesion rats, which may be related to the regulation of FIB, sICAM-1, TGF-β_1 level. It can improve the endothelial function and the inhibition of inflammatory response.