Jianpi Qingre Huoxue decoction treating ulcerative colitis by PGRP-3 and PGRP-4 and IL-22
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摘要: [目的]基于检测溃疡性结肠炎肽聚糖结合蛋白3(PGRP-3)、肽聚糖结合蛋白4(PGRP-4)、IL-22表达及结肠黏膜组织病理学变化,探讨健脾清热活血方干预溃疡性结肠炎的可能机制。[方法]100只Balb-c小鼠按体重随机分为5组即正常组、模型组、低剂量组、中剂量组及高剂量组,每组20只,采用自由引用3%DSS法制备溃疡性结肠炎模型,自第3周起分组给药,模型组予等剂量生理盐水、治疗组分别予不同剂量健脾清热活血方灌胃,连续1周。末次给药后处死全部小鼠,采用苏木精-伊红染色法检测小鼠结肠组织病理学变化,采用Western-blot技术检测结肠黏膜PGRP-3及PGRP-4表达,应用ELISA检测IL-22表达。[结果]光镜下见模型组结肠黏膜充血、肿胀,黏膜固有层及黏膜下层大量炎性细胞浸润,见溃疡形成;而治疗组肠壁黏膜及黏膜下层轻度充血水肿,少量炎细胞浸润,未见明显溃疡。模型组PGRP-3、PGRP-4表达下降;治疗组PGRP-3、PGRP-4表达上升,经比较,差异有统计学意义(P<0.05)。模型组IL-22表达量为(398.67±68.76)pg/ml,治疗组中低剂量组IL-22表达量为(257.82 ±56.41)pg/ml、中剂量组为(249.32±61.75)pg/ml、高剂量组为(263.69±57.89)pg/ml,治疗组与模型组比较有统计学意义(P<0.05)。[结论]健脾清热活血方可能通过调控PGRP识别PGN,杀伤外源微生物,传递炎症信号,促进IL-22分泌增加,增强上皮先天防御机制和屏障的完整性,隔离免疫系统与肠道共生菌群之间的直接相互作用,保护肠道免受炎症损伤,从而发挥缓解UC的效用。Abstract: [Objective] To explore the mechanism of Jianpi Qingre Huoxue decoction(JQHD)in the treatment of ulcerative colitis by observing the expression of PGRP-3,PGRP-4 and IL-22.[Methods] 100 Balb-c mice were randomly divided into normal group,model group,low dosage group,middle dosage group and high dosage group,20 mice in each group.The ulcerative colitis models were made by drinking 3% DSS for 2 weeks.Since the 3 th week,the model group was given equal dose of saline gavage.The latter three groups were treated with different dosage of JQHD gavage for one week.All rats were sacrificed after the last administration.The pathological changes of colonic mucosa were detected by HE staining.The expression of PGRP-3 and PGRP-4 in colonic mucosa was detected by Western-blot and IL-22 with ELISA respectively.[Results] The model group showed colonic mucosa hyperemia swelling,a large number of inflammatory cell infiltration and ulcers in mucosa lamina propria and submucosa under the light microscope.While the intervention groups showed improved inflammatory cell infiltration with no noticeable ulcer and low-grade hyperemia swelling.In the model group,the expression of PGRP-3 and PGRP-4 decreased.The expression of PGRP-3 and PGRP-4 in the JQHD groups increased,with significant difference as compared with the model group(P<0.05).The expression of IL-22 was 398.67±68.76 pg/ml,257.82 ±56.41 pg/ml,249.32±61.75 pg/ml and 263.69±57.89 pg/ml in the model group,low dosage,middle dosage,high dosage JQHD group respectively.Compared with the model group,JQHD groups showed significant difference(P<0.05).[Conclusion] JQHD decoction may exert a distinct effect on improving ulcerative colitis by regulating PGRP sensed PGN,eliminating the exogenous microorganisms,mediating the inflammatory signals,resulting in the promotion of IL-22 secretion,enhancement of the integrity of the epithelium and the protection of the immune system and the direct interaction between the immune system and intestinal symbiotic bacteria,so as to protect the intestine from inflammatory injury.
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