理冲生髓饮有效组分调控炎性因子对卵巢癌侵袭转移影响的研究

Study on the effect of effective components of Lichong Shengsui Decoction on inflammatory factors in invasion and metastasis of ovarian cancer

目的探究理冲生髓饮有效组分通过调控炎性因子对卵巢癌SKOV3细胞侵袭转移的影响。方法①将32只大鼠随机分为空白组、生髓饮低剂量组、生髓饮中剂量组和生髓饮高剂量组,每组8只。生髓饮低、中、高剂量组分别给予3.76 g/(kg·d)、7.52 g/(kg·d)、11.28 g/(kg·d)理冲生髓饮灌胃,空白组给予同体积蒸馏水灌胃,均2次/d,连续7 d。末次灌胃结束2 h后眼眶取血,获得含药血清。②实验分为空白组、生髓饮低剂量含药血清组、生髓饮中剂量含药血清组、生髓饮高剂量含药血清组和贝伐单抗组,应用鸡胚绒毛尿囊膜血管实验观察各组血管生成情况。③选取在体外培养至对数生长期的人卵巢癌SKOV3细胞,分为空白组、生髓饮低剂量含药血清组、生髓饮中剂量含药血清组、生髓饮高剂量含药血清组和贝伐单抗组,除空白组外,其余组分别加入相应浓度的理冲生髓饮含药血清及5 mg/L贝伐单抗培养,应用MTT法观察各组细胞增殖情况,用Transwell小室实验以及划痕实验检测细胞的迁移和侵袭能力,应用ELISA法检测卵巢癌相关炎性因子蛋白表达水平。结果生髓饮各含药血清组及贝伐单抗组一级和二级血管数目均明显少于空白组(P均<0.05),且生髓饮中剂量含药血清组和贝伐单抗组均明显少于生髓饮低、高剂量含药血清组(P均<0.05);与空白组比较,生髓饮各含药血清组及贝伐单抗组SKOV3细胞迁移率和侵袭率均显著降低(P均<0.05),且生髓饮中、高剂量含药血清组和贝伐单抗组均显著低于生髓饮低剂量含药血清组(P均<0.05)。生髓饮各含药血清组及贝伐单抗组血管内皮生长因子(VEGF)、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)水平及生髓饮高剂量含药血清组、贝伐单抗组白细胞介素-1(IL-1)水平均明显低于空白组(P均<0.05);生髓饮高剂量含药血清组和贝伐单抗组VEGF、IL-6、TNF-α水平均明显低于生髓饮中、低剂量含药血清组(P均<0.05),各组基质金属蛋白酶-9(MMP-9)水平比较差异均无统计学意义(P均>0.05)。结论理冲生髓饮有效组分能够抑制卵巢癌SKOV3细胞的侵袭转移,其作用机制与抑制相关炎性因子的表达、影响肿瘤周围的炎性微环境有关。

Objective It is to investigate the effect of effective components of Lichong Shengsui Decoction on invasion and metastasis of ovarian cancer SKOV3 cells by regulating inflammatory factors.Methods①Thirty-two rats were randomly divided into blank group,low-dose Lichong Shengsui Decoction group,medium-dose Lichong Shengsui Decoction group and high-dose Lichong Shengsui Decoction group,with 8 rats in each group.the low,medium and high dose groups of Lichong Shengsui Decoction were respectively given 3.76 g/(kg·d),7.52 g/(kg·d),11.28 g/(kg·d)Lichong Shengsui Decoction by gavage,the blank group was given the same volume of distilled water by gavage,all 2 times/d,for 7 consecutive days.The blood was taken from the eye socket in 2 hours after the last gavage to obtain medicated serum.②The experiment was divided into blank group,low-dose Lichong Shengsui Decoction medicated serum group,medium-dose Lichong Shengsui Decoction medicated serum group,high-dose Lichong Shengsui Decoction medicated serum group and bevacizumab group,The chicken embryo chorioallantoic membrane vascular experiment was used to observe the angiogenesis of each group.③The human ovarian cancer SKOV3 cells cultured in vitro to logarithmic growth phase were selected and divide into blank group,low-dose Lichong Shengsui Decoction medicated serum group,medium-dose Lichong Shengsui Decoction medicated serum group,high-dose Lichong Shengsui Decoction medicated serum group and bevacizumab group.Except for the blank group,the other groups were cultured with the corresponding concentration of Lichong Shengsui Decoction medicated serum and 5 mg/L bevacizumab.The cell proliferation of each group was observed by the MTT method,the cell migration and invasion ability were detected by Transwell chamber test and scratch test,the expression level of inflammatory factor protein related to ovarian cancer was detected by ELISA.Results The number of primary and secondary blood vessels in each drug-containing serum group of Lichong Shengsui Decoction and bevacizumab group was significantly less than that of the blank group(P<0.05),and the medium dose Lichong Shengsui Decoction drug-containing serum group and bevacizumab group were significantly less than that in the low and high-dose drug-containing serum groups(P<0.05).The SKOV3 cell migration rate and invasion rate was significantly reduced in each drug-containing serum group of Lichong Shengsui Decoction and bevacizumab group compared with the blank group(all P<0.05),and the middle and high-dose drug-containing serum group of Lichong Shengsui Decoction and the bevacizumab group were significantly lower than those of the low-dose drug-containing serum group(all P<0.05).The levels of vascular endothelial growth factor(VEGF),interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)in each drug-containing serum group of Lichong Shengsui Decoction and bevacizumab group and the levels of interleukin-1(IL-1)in the high-dose Shengsui Decoction drug-containing serum group and bevacizumab group were significantly lower than those in the blank group(all P<0.05);the levels of VEGF,IL-6 and TNF-αin the high-dose Lichong Shengsui Decoction drug-containing serum group and bevacizumab group were significantly lower than those in the medium and low-dose Lichong Shengsui Decoction drug-containing serum groups(all P<0.05),there was no significant difference in the levels of matrix metalloproteinase-9(MMP-9)between every group(all P>0.05).Conclusion The effective components of Lichong Shengsui Decoction can inhibit the invasion and metastasis of ovarian cancer SKOV3 cells,and its mechanism of action is related to inhibiting the expression of related inflammatory factors and affecting the inflammatory microenvironment around the tumor.