扶正消萎汤对N-甲基-N'-硝基-N-亚硝基胍诱导的慢性萎缩性胃炎大鼠胃黏膜修复及胃肠道动力的影响研究

doi:10.3877/cma.j.issn.2095-8765.2023.01.003

目的

研究扶正消萎汤对N-甲基-N'-硝基-N-亚硝基胍(MNNG)诱导的慢性萎缩性胃炎(CAG)大鼠胃黏膜修复及胃肠道动力的影响。

方法

大鼠随机分为正常对照组及造模组，采用MNNG联合乙醇灌胃法建立CAG大鼠模型，造模完成后大鼠随机分为模型组、扶正消萎汤低、中、高剂量组及维酶素组，每组10只，扶正消萎汤低、中、高剂量组分别按照0.74、1.48、2.22 g/kg(以生药含量计) 灌胃给予大鼠的扶正消萎汤，维酶素组按照240 mg/kg灌胃给予维酶素，1次/d，连续6周，测定大鼠胃排空率及小肠推进比，采用酶联免疫吸附(ELISA)试剂盒测定大鼠血清胃泌素(GAS)、胃动素(MTL)及生长抑素(SS)水平及大鼠胃黏膜组织内皮素(ET)、前列腺素E2(PGE2)、分泌型免疫球蛋白A (sIgA)、还原型谷胱甘肽(GSH)水平，苏木精-伊红(HE)染色法检查胃黏膜组织病理学变化，缺口末端标记法(TUNEL)染色法检查大鼠胃黏膜组织细胞凋亡率，免疫印迹法测定大鼠胃黏膜组织核转录因子2(Nrf2)、血红素氧合酶1(HO-1)蛋白水平。

结果

与正常对照组比较，模型组大鼠血清GAS、SS水平、胃排空率及小肠推进比、胃黏膜PGE2、sIgA、GSH水平、胃黏膜组织Nrf2、HO-1蛋白水平显著降低(P<0.05)，血清MTL水平、胃黏膜组织ET水平、胃黏膜组织细胞凋亡率显著升高(P<0.05)；与模型组比较，扶正消萎汤各剂量组及维酶素组大鼠血清GAS、SS水平、胃排空率及小肠推进比、胃黏膜PGE2、sIgA、GSH水平、胃黏膜组织Nrf2、HO-1蛋白水平显著升高(P<0.05)，血清MTL水平、胃黏膜组织ET水平、胃黏膜组织细胞凋亡率显著降低(P<0.05)，且随着扶正消萎汤剂量的增加，上述指标变化更明显。

结论

扶正消萎汤能够显著调节CAG大鼠胃肠道激素分泌，抑制胃黏膜细胞凋亡，促进胃动力恢复，修复胃黏膜病理改变，其机制可能与调节Nrf2/HO-1通路有关。

关键词: 扶正消萎汤, 慢性萎缩性胃炎, 胃肠道动力

Objective

To study the effects of Fuzheng Xiaowei Decoction on gastrointestinal motility and gastric mucosal repair in chronic atrophic gastritis (CAG) rats induced by N-Methyl-N'-nitro-N-nitrosoguanidine(MNNG).

Methods

The rats were randomly divided into normal control group and modeling group, and the CAG rat model was established by MNNG combined with ethanol gavage. After modeling, the rats were randomly divided into model group, Fuzheng Xiaowei decoction low-dose, medium-dose and high-dose groups and enzyme group (10 rats per group). The low, medium and high dose groups of FuzhengXiaowei Decoction were given 0.74 g/kg, 1.48 g/kg and 2.22 g/kg by intragastric administration, respectively, and the vitase group was given 240 mg/kg intragastric administration of vitase, once a day for 6 w, and the gastric empting rate and small intestine propulsive ratio of rats were measured. The levels of gastrin (GAS), motilin (MTL) and somatostatin (SS) in rat serum and endothelin (ET), Prostaglandin E2 (PGE2), Secretory immunoglobulin A (sIgA) and glutathione (GSH) in rat gastric mucosa were determined by enzyme-linked immunosorbent assay (ELISA). The histopathological changes of gastric mucosa were detected by hematoxylin-eosin (HE) staining, and the apoptosis rate of gastric mucosal tissue was detected by Td-mediated dudpo-x nick end Labeling (TUNEL) staining. The protein levels of nuclear tran-scription factor 2 (Nrf2) and heme oxygenose 1 (HO-1) in gastric mucosa of rats were determined by Western blot.

Results

Compared with the normal control group, the levels of serum GAS, SS, gastric emptier rate, intestinal propulsion ratio, PGE2, sIgA, GSH levels, Nrf2 and HO-1 protein levels of gastric mucosa in model group were significantly decreased (P<0.05). Serum MTL level, ET level and apoptosis rate of gastric mucosa tissue were significantly increased (P<0.05). Compared with model group, the serum GAS, SS levels, gastric emptying rate, small intestine advancement ratio, PGE2, sIgA, GSH levels, Nrf2 and HO-1 protein levels of gastric mucosa in FuzhengXiaowei Decoction and vitase group were significantly increased (P<0.05). Serum MTL level, ET level in gastric mucosa tissue and apoptosis rate of gastric mucosa tissue were significantly decreased (P<0.05), and the above indexes were more significantly changed with the increase of the dose of FuzhengXiaowei Decoction.

Conclusion

Fuzheng Xiaowei Decoction can significantly regulate gastrointestinal hormone secretion, inhibit the apoptosis of gastric mucosa cells, promote the recovery of gastric motility, and repair the pathological changes of gastric mucosa in rats with chronic atrophic gastritis. The mechanism may be related to the regulation of Nrf2/HO-1 pathway.

Key words: Fuzheng Xiaowei Decoction, CAG rats, Gastrointestinal motility

表1 大鼠血清GAS、MTL及SS水平比较结果（ng/L，）

组别	GAS	MTL	SS
正常对照组	1361.29±24.20	149.88±2.05	79.02±2.14
模型组	813.05±18.77^a	242.53±3.98^a	43.62±1.91^a
扶正消萎汤低剂量组	947.43±14.62^b	193.19±1.79^b	58.78±1.47^b
扶正消萎汤中剂量组	1183.71±26.46^bc	178.77±2.67^bc	65.05±1.82^bc
扶正消萎汤高剂量组	1289.54±26.34^bcd	155.80±3.58^bcd	74.71±1.39^bcd
维酶素组	1178.75±17.55	161.13±3.52	72.33±2.52

表1 大鼠血清GAS、MTL及SS水平比较结果（ng/L，）

组别	GAS	MTL	SS
正常对照组	1361.29±24.20	149.88±2.05	79.02±2.14
模型组	813.05±18.77^a	242.53±3.98^a	43.62±1.91^a
扶正消萎汤低剂量组	947.43±14.62^b	193.19±1.79^b	58.78±1.47^b
扶正消萎汤中剂量组	1183.71±26.46^bc	178.77±2.67^bc	65.05±1.82^bc
扶正消萎汤高剂量组	1289.54±26.34^bcd	155.80±3.58^bcd	74.71±1.39^bcd
维酶素组	1178.75±17.55	161.13±3.52	72.33±2.52

表2 大鼠胃排空率及小肠推进比比较结果（%，）

组别	胃排空率	小肠推进比
正常对照组	81.23±3.64	63.14±2.25
模型组	53.04±1.57^a	40.09±1.64^a
扶正消萎汤低剂量组	62.26±1.17^b	49.21±1.92^b
扶正消萎汤中剂量组	70.84±2.96^bc	55.27±1.44^bc
扶正消萎汤高剂量组	78.29±3.36^bcd	60.78±1.26^bcd
维酶素组	75.20±1.20^b	57.26±1.55^b

表2 大鼠胃排空率及小肠推进比比较结果（%，）

组别	胃排空率	小肠推进比
正常对照组	81.23±3.64	63.14±2.25
模型组	53.04±1.57^a	40.09±1.64^a
扶正消萎汤低剂量组	62.26±1.17^b	49.21±1.92^b
扶正消萎汤中剂量组	70.84±2.96^bc	55.27±1.44^bc
扶正消萎汤高剂量组	78.29±3.36^bcd	60.78±1.26^bcd
维酶素组	75.20±1.20^b	57.26±1.55^b

表3 大鼠胃黏膜组织ET、PGE2、sIgA、GSH水平比较结果（）

组别	ET/(ng/L)	PGE2/(ng/L)	sIgA/(μg/ml)	GSH/(μmol/L)
正常对照组	240.19±2.71	83.96±2.24	63.44±3.77	64.89±2.94
模型组	389.71±3.69^a	37.14±1.89^a	20.23±3.78^a	8.23±1.36^a
扶正消萎汤低剂量组	318.07±3.41^b	51.29±1.93^b	35.07±2.47^b	38.45±2.70^b
扶正消萎汤中剂量组	264.72±1.58^bc	62.77±3.12^bc	48.66±3.31^bc	47.49±2.63^bc
扶正消萎汤高剂量组	248.06±2.28^bcd	79.11±2.13^bcd	57.07±4.69^bcd	58.97±2.60^bcd
维酶素组	253.61±2.63^b	73.54±2.67^b	55.23±1.58^b	55.33±1.69^b

表3 大鼠胃黏膜组织ET、PGE2、sIgA、GSH水平比较结果（）

组别	ET/(ng/L)	PGE2/(ng/L)	sIgA/(μg/ml)	GSH/(μmol/L)
正常对照组	240.19±2.71	83.96±2.24	63.44±3.77	64.89±2.94
模型组	389.71±3.69^a	37.14±1.89^a	20.23±3.78^a	8.23±1.36^a
扶正消萎汤低剂量组	318.07±3.41^b	51.29±1.93^b	35.07±2.47^b	38.45±2.70^b
扶正消萎汤中剂量组	264.72±1.58^bc	62.77±3.12^bc	48.66±3.31^bc	47.49±2.63^bc
扶正消萎汤高剂量组	248.06±2.28^bcd	79.11±2.13^bcd	57.07±4.69^bcd	58.97±2.60^bcd
维酶素组	253.61±2.63^b	73.54±2.67^b	55.23±1.58^b	55.33±1.69^b

图1 大鼠胃黏膜组织病理学检查结果注：A为正常对照组；B为模型组；C为扶正消萎汤低剂量组；D为扶正消萎汤中剂量组；E为扶正消萎汤高剂量组；F为维酶素组

图2 大鼠胃黏膜组织细胞凋亡率比较结果注：A为正常对照组；B为模型组；C为扶正消萎汤低剂量组；D为扶正消萎汤中剂量组；E为扶正消萎汤高剂量组；F为维酶素组。与正常对照组比较，^aP<0.05；与模型组比较，^bP<0.05；与扶正消萎汤低剂量组比较，^cP<0.05；与扶正消萎汤中剂量组比较，^dP<0.05

图3 大鼠胃黏膜组织Nrf2、HO-1蛋白水平注：A为正常对照组；B为模型组；C为扶正消萎汤低剂量组；D为扶正消萎汤中剂量组；E为扶正消萎汤高剂量组；F为维酶素组。

表4 大鼠胃黏膜组织Nrf2、HO-1蛋白水平比较结果

组别	Nrf2	HO-1
空白对照组	0.66±0.25	0.53±0.27
模型组	0.11±0.07^a	0.17±0.06^a
扶正消萎汤低剂量组	0.25±0.09^b	0.29±0.08^b
扶正消萎汤中剂量组	0.34±0.07^bc	0.36±0.17^bc
扶正消萎汤高剂量组	0.64±0.15^bcd	0.44±0.09^bcd
阳性对照组	0.26±0.08^b	0.26±0.08^b

表4 大鼠胃黏膜组织Nrf2、HO-1蛋白水平比较结果

组别	Nrf2	HO-1
空白对照组	0.66±0.25	0.53±0.27
模型组	0.11±0.07^a	0.17±0.06^a
扶正消萎汤低剂量组	0.25±0.09^b	0.29±0.08^b
扶正消萎汤中剂量组	0.34±0.07^bc	0.36±0.17^bc
扶正消萎汤高剂量组	0.64±0.15^bcd	0.44±0.09^bcd
阳性对照组	0.26±0.08^b	0.26±0.08^b

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Effect of Fuzheng Xiaowei Decoction on gastrointestinal motility and gastric mucosa repair in chronic atrophic gastritis rats induced by N-Methyl-N'-nitro-N-nitrosoguanidine

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Effect of Fuzheng Xiaowei Decoction on gastrointestinal motility and gastric mucosa repair in chronic atrophic gastritis rats induced by N-Methyl-N'-nitro-N-nitrosoguanidine