加味葛根芩连汤对湿热泄泻仔猪肠道炎症和损伤修复的作用

通过用加味葛根芩连汤治疗仔猪的湿热泄泻，探讨其对湿热泄泻仔猪肠道的修复作用。选取8~15日龄未断奶仔猪（大白×长白）30头，其中健康仔猪6头，为对照组；有粪色黄褐而臭、小便短赤、舌质红等症状的泄泻仔猪24头，分为湿热泄泻组、加味葛根芩连汤低剂量组、加味葛根芩连汤中剂量组和加味葛根芩连汤高剂量组，每组6头。对照组和湿热泄泻组仔猪灌服生理盐水，药物组仔猪灌服加味葛根芩连汤（按生药量，低、中、高剂量组药物添加量分别为2.5、5.0和10.0 g·d^-1），连续5 d。在服药的第0、3、5天记录各组仔猪的粪便评分。取对照组、湿热泄泻组、加味葛根芩连汤中剂量组仔猪十二指肠、空肠、回肠和结肠进行组织切片和H.E.染色，取十二指肠进行增殖性细胞核抗原（PCNA）免疫组化染色；用荧光定量PCR检测对照组、湿热泄泻组、加味葛根芩连汤中剂量组仔猪空肠TLR4、TNF-α、IL-1β和IL-6 mRNA表达量变化。湿热泄泻严重破坏了仔猪的十二指肠、空肠、回肠和结肠结构，尤其是十二指肠和空肠，十二指肠隐窝处PCNA表达量显著升高（P<0.05）。与对照组相比，湿热泄泻组仔猪空肠TLR4（P<0.05）、TNF-α（P<0.01）、IL-1β（P<0.01）和IL-6（P<0.05）mRNA表达量显著升高。相比于湿热泄泻组，加味葛根芩连汤显著降低了仔猪的粪便评分（P<0.05）。中剂量加味葛根芩连汤组仔猪的小肠和结肠的组织结构得到显著改善，十二指肠的肠腔中有不断脱落的绒毛团，绒毛中的PCNA表达量极显著升高（P<0.01）。与湿热泄泻组相比，加味葛根芩连汤中剂量组空肠TLR4（P<0.05）、TNF-α（P<0.01）、IL-1β（P<0.01）和IL-6（P<0.05）mRNA表达量显著降低。综上表明，加味葛根芩连汤可通过促进隐窝干细胞的增殖和降低炎症反应对湿热泄泻造成的仔猪肠道损伤进行修复。

Effect of Jiawei Gegen Qinlian Decoction on the Regulation of Intestine Damage and Repair in the Piglets of Damp-heat Diarrhea

The purpose of the study was to research the effect of Jiawei Gegen Qinlian Decoction (JGQLD) on the regulation of intestine damage and repair in the piglets of damp-heat diarrhea (DHD). Six piglets (8-15 days old) were selected as control group. Twenty-four piglets (8-15 days old) that with the symptom of DHD were divided into 4 groups, including DHD group, low dose, middle dose and high dose of JGQLD group, 6 piglets per group. Piglets in the control group and the DHD group were given normal saline. Piglets in medicine groups were given the JGQLD for 5 consecutive days (the doses were recorded as dried medicinal herb, 2.5 g·d^-1 in the low dose group, 5.0 g·d^-1 in the middle dose, and 10.0 g·d^-1 in the high dose group). The fecal scores of piglets in all groups were recorded at 0, 3 and 5 day. In control group, DHD group and middle dose group of JGQLD, the duodenum, jejunum, ileum and colon of piglets were taken for tissue section and H.E. staining, while the duodenum was selected for proliferating cell nuclear antigen (PCNA) immune-histochemical staining. The mRNA expressions of TLR4, TNF-α, IL-1β and IL-6 in the jejunum of piglets from control group, DHD group and middle dose of JGQLD group were detected by real time PCR. In the DHD group, DHD destroyed the structure of small intestine and colon, especially the duodenum and jejunum. Compared with the control group, the PCNA expression in the duodenum crypt of the DHD group was significantly increased (P<0.05), the mRNA expressions of TLR4 (P<0.05), TNF-α (P<0.01), IL-1β (P<0.01) and IL-6 (P<0.05) in the jejunum of the DHD group were all increased significantly. Compared with the DHD group, the low, middle and high dose of JGQLD significantly decreased the fecal scores of piglets (P<0.05). The structures of duodenum, jejunum, ileum and colon in the middle dose of JGQLD group were improved and there were a few shedding villus in the lumen of duodenum. Compare with the DHD group, the PCNA expression in the duodenum villus of the middle dose group was extremely significantly increased (P<0.01), the mRNA expressions of TLR4 (P<0.05), TNF-α (P<0.01), IL-1β (P<0.01) and IL-6 (P<0.05) in the jejunum of the middle dose group were significantly decreased. JGQLD repaired the intestine damage of piglets with DHD by promoting the proliferation of crypt stem cells and reducing the intestinal inflammatory response.