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1.上海中医药大学 附属龙华医院,上海 200032
2.福建中医药大学 附属第二人民医院,福州 350003
Received:06 October 2024,
Revised:2024-11-26,
Accepted:17 February 2025,
Online First:18 February 2025,
Published:05 June 2026
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何友成,蒋风茹,吴月等.大黄酸诱导的泻药性结肠大鼠模型气阴两虚伴气滞血瘀证候分析[J].中国实验方剂学杂志,2026,32(11):185-195.
HE Youcheng,JIANG Fengru,WU Yue,et al.Qi and Yin Deficiency with Qi Stagnation and Blood Stasis in Rat Model of Rhein-induced Cathartic Colon[J].Chinese Journal of Experimental Traditional Medical Formulae,2026,32(11):185-195.
何友成,蒋风茹,吴月等.大黄酸诱导的泻药性结肠大鼠模型气阴两虚伴气滞血瘀证候分析[J].中国实验方剂学杂志,2026,32(11):185-195. DOI: 10.13422/j.cnki.syfjx.20250314.
HE Youcheng,JIANG Fengru,WU Yue,et al.Qi and Yin Deficiency with Qi Stagnation and Blood Stasis in Rat Model of Rhein-induced Cathartic Colon[J].Chinese Journal of Experimental Traditional Medical Formulae,2026,32(11):185-195. DOI: 10.13422/j.cnki.syfjx.20250314.
目的
2
评价并分析大黄酸诱导的泻药性结肠(CC)大鼠模型的气阴两虚伴气滞血瘀证候。
方法
2
将24只大鼠分为正常组和模型组(CC组),分别饲以等体积的生理盐水和2%大黄酸混悬液,进行为期3个循环约118 d的造模:第一循环46 d,12 mL·kg
-1
·d
-1
,隔日灌胃;第二循环37 d,12 mL·kg
-1
·d
-1
,灌5 d停2 d;第三循环35 d,16 mL·kg
-1
·d
-1
,灌5 d停2 d。每个循环以80%的大鼠稀便消失为每个循环结束标准。观测大鼠体质量、24 h进食量、皮毛及其红(R)、绿(G)、蓝(B)值,旷场实验(OFT)测量大鼠活动总路程,以评定气虚;观测大鼠体质量系数、24 h饮水量,以评定阴虚;糖水偏好实验(SPT)测定糖水偏好度(SPR),OFT测量大鼠平均速度,评定抑郁状态(肝郁气滞);观测舌象及其R、G、B值,全自动血液流变仪器测定全血黏度和血浆黏度,评定血瘀;碳墨推进实验测量肠道推进率(ITR),以评定疾病模型;苏木素-伊红(HE)染色观察大鼠结肠病理组织学情况;实时荧光定量聚合酶链式反应(Real-time PCR)检测结肠组织瞬时受体电位锚蛋白1(TRPA1)和色氨酸羟化酶1(TPH1)的mRNA表达;蛋白免疫印迹法(Western blot)检测结肠组织TRPA1、TPH1蛋白表达情况。
结果
2
证候指标方面,与正常组比较,CC组体质量明显降低(
P
<
0.05);24 h进食量显著增加(
P
<
0.01),且被毛萎黄,凌乱欠华,被毛R、G、B值均显著降低(
P
<
0.01),OFT总路程显著减少(
P
<
0.01);体质量系数显著降低(
P
<
0.01),24 h饮水量明显增加(
P
<
0.05,
P
<
0.01);SPR显著降低(
P
<
0.01),OFT平均速度显著减慢(
P
<
0.01);舌质暗红,舌象R、G、B值均显著降低(
P
<
0.01),全血黏度和血浆黏度显著升高(
P
<
0.01)。疾病指标方面,与正常组比较,CC组ITR减慢(
P
<
0.01)。病理组织学上,HE染色可见CC组结肠黏膜上皮细胞坏死、脱落,结肠黏膜连续性受到破坏,伴见固有层炎性细胞浸润;半定量结果显示HAI评分明显升高(
P
<
0.05)、炎性细胞计数及其面积占比明显增多(
P
<
0.05)。分子生物学指标上,与正常组比较,CC组结肠组织TRPA1、TPH1的mRNA和蛋白表达水平明显降
低(
P
<
0.05,
P
<
0.01)。
结论
2
大黄酸诱导的CC大鼠模型符合气阴两虚伴气滞血瘀的中医证候特征。
Objective
2
To evaluate and analyze the syndrome characteristics of Qi and Yin deficiency accompanied by Qi stagnation and blood stasis in a rhein-induced cathartic colon (CC) rat model.
Methods
2
Twenty-four rats were divided into a normal group and a model group (CC group). The rats were administered equal volumes of physiological saline or 2% rhein suspension by gavage to establish the model over three cycles (approximately 118 days). The first cycle lasted 46 days, with a dosage of 12 mL·kg
-1
·d
-1
, administered every other day. The second cycle lasted 37 days, with a dosage of 12 mL·kg
-1
·d
-1
, administered for 5 consecutive days followed by 2 days of cessation. The third cycle lasted 35 days, with a dosage of 16 mL·kg
-1
·d
-1
, also administered for 5 consecutive days followed by 2 days of cessation. Each cycle ended when 80% of the rats no longer exhibited loose stools. Body mass, 24 h food intake, coat condition, and coat red (R), green (G), and blue (B) values were recorded. The open field test (OFT) was used to measure the total distance traveled to evaluate Qi deficiency. The body mass coefficient and 24 h water intake were recorded to assess Yin deficiency. The sucrose preference test (SPT) was used to determine the sucrose preference rate (SPR), and the average speed in OFT was measured to evaluate depressive status (liver depression and Qi stagnation). Tongue images and their R, G, and B values were recorded. Whole blood viscosity (WBV) and plasma viscosity (PV) were measured using an automatic hemorheological analyzer to evaluate blood stasis. A carbon ink propulsion test was performed to determine the intestinal transit rate (ITR) for disease model evaluation. Hematoxylin-eosin (HE) staining wa
s used to observe histopathological changes in the colon. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was used to detect the mRNA expression of transient receptor potential ankyrin 1 (TRPA1) and tryptophan hydroxylase 1 (TPH1) in colon tissue. Western blot was used to detect the protein expression of TRPA1 and TPH1.
Results
2
In terms of syndrome indicators, compared with the normal group, the body mass of the CC group decreased (
P
<
0.05), while 24 h food intake increased (
P
<
0.01). The coats of the CC group appeared withered, disheveled, and dull, and the R, G, and B values of the coat decreased (
P
<
0.01). The total distance traveled in OFT decreased (
P
<
0.01). The body mass coefficient decreased (
P
<
0.01), while 24 h water intake increased (
P
<
0.05,
P
<
0.01). The SPR decreased (
P
<
0.01), and the average speed in OFT slowed (
P
<
0.01). The tongue appeared dark red, and the R, G, and B values of tongue images decreased (
P
<
0.01). WBV and PV increased (
P
<
0.01). Regarding disease indicators, compared with the normal group, the ITR decreased in the CC group (
P
<
0.01). Pathologically, HE staining showed necrosis and shedding of colonic mucosal epithelial cells, disruption of mucosal continuity, and infiltration of inflammatory cells in the lamina propria in the CC group. Semi-quantitative analysis showed increased HAI scores (
P
<
0.05) and increased inflammatory cell counts and area proportion (
P
<
0.05). In terms of molecular biological indicators, compared with the normal group, the mRNA and protein expression levels of TRPA1 and TPH1 in colon tissue decreased in the CC group (
P
<
0.05,
P
<
0.01).
Conclusion
2
The rhein-induced CC rat model conforms to the traditional Chinese medicine syndrome characteristics of Qi and Yin deficiency accompanied by Qi stagnation and blood stasis.
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