“益氣排毒方”治療慢性腎功能不全的實驗研究
發(fā)布時間：2018-08-29 18:05
【摘要】：慢性腎功能不全又稱慢性腎臟病,是以代謝產(chǎn)物潴留,水、電解質(zhì)、酸堿平衡失調(diào),全身各系統(tǒng)受累為主要表現(xiàn)的臨床綜合征。隨著醫(yī)學(xué)的進步和檢驗方法和技術(shù)的不斷提高,慢性腎功能不全的發(fā)病率逐年升高。許多慢性腎功能不全患者由于治療不及時、不恰當(dāng),導(dǎo)致體內(nèi)毒素蓄積,增加對腎臟功能的損害,從而迅速進入尿毒癥期。尿毒癥毒素蓄積,嚴(yán)重危害患者的生活和健康,降低生活質(zhì)量,龐大的治療費用更是給家庭和社會帶來巨大的負(fù)擔(dān)。所以,探索治療慢性腎功能不全的新方法非常有臨床意義。 益氣排毒方是清遠(yuǎn)市中醫(yī)院歐陽永紅教授30余年臨床經(jīng)驗的總結(jié),主要應(yīng)用于治療有腎功能損害臨床表現(xiàn),實驗室指標(biāo)血尿素氮、肌酐升高的患者,即慢性腎功能不全的病人,收入清遠(yuǎn)市中醫(yī)院協(xié)定處方,在改善患者精神狀況、貧血、四肢乏力等癥狀,降低血肌酐、尿素氮、蛋白尿、尿液潛血等方面效果顯著。1992年歐陽永紅教授提出使用“益氣瀉濁”法治療慢性腎功能不全,2011年王瑞等研究發(fā)現(xiàn)益氣排毒方對維持性血液透析患者的微炎癥狀態(tài)有干預(yù)作用,可降低微炎癥狀態(tài)對機體的損害,改善營養(yǎng)不良；2012年“益氣排毒方治療慢性腎功能不全脾腎氣虛證的臨床研究”已列為廣東省中醫(yī)藥管理局的課題。為了闡明益氣排毒方改善慢性腎功能不全的機理,我們通過動物實驗,論證益氣排毒方在改善大鼠一般情況,保護大鼠腎功能,調(diào)節(jié)血脂和鈣磷代謝,改善貧血等作用,并嘗試探討方劑的作用機理。 目的： 采用腺嘌呤毒害大鼠腎臟,建立慢性腎功能不全大鼠模型,觀察益氣排毒方對慢性腎功能不全大鼠一般情況的影響,對血肌酐、尿素氮、血紅蛋白、血脂、血液鈣磷代謝等指標(biāo)的影響,并觀察腎臟病理改變。對比尿毒清顆粒對大鼠腎臟保護作用,分析作用機制,探討益氣排毒方糾正慢性腎衰竭的作用機理。 方法： 選取180-220gSPF級wistar大鼠40只,雌雄各半,給予固體飲食和自由飲水,規(guī)律喂養(yǎng)一周后,隨機分為4組,每組10只,選取一組為正常對照組,其余30只為慢性腎功能不全造模組,給予100mg/kg/d腺嘌呤灌胃,連續(xù)喂養(yǎng)21天造模成功后,選取造模成功的大鼠,按照體重大小重新排序,按照計算機產(chǎn)生的隨機數(shù)字將已造模的大鼠,隨機分配為三組：模型對照組、尿毒清組、益氣排毒方組,根據(jù)正常成人和大鼠體重?fù)Q算,其中尿毒清組按照按照4.0g/kg的比例,灌胃；益氣排毒方組按照23.38ml/kg來灌胃,平均每只大鼠約4ml藥液,連續(xù)給藥28天。麻醉,取血,檢測血肌酐、尿素氮、血脂、電解質(zhì)等指標(biāo),觀察雙腎病理改變。 結(jié)果： 1、造模結(jié)果：造模后,造模組大鼠與正常組大鼠相比,精神狀態(tài)差,反應(yīng)遲鈍,毛色灰白,活動減少。體重與正常組相比,差異有統(tǒng)計學(xué)意義(P0.05)；但造模組之間,體重?zé)o顯著差異(P0.05)。隨機抽取一只造模大鼠,心臟取血,檢驗?zāi)I功能提示,血肌酐和尿素氮顯著升高,腎臟病理解剖見腎臟體積增大,顏色淡白,呈現(xiàn)“大白腎”,HE染色見腎小球、腎小管、腎間質(zhì)部分組織結(jié)構(gòu)損壞。 2、治療結(jié)果： (1)一般情況。灌服益氣排毒方的慢性腎功能不全大鼠,一般情況優(yōu)于模型對照組,大鼠精神狀態(tài)好,反應(yīng)敏捷,進食量多,毛發(fā)較有光澤,但尚未達到正常水平；與尿毒清組相比,益氣排毒方組大鼠與其精神狀態(tài)、反應(yīng)靈敏度相當(dāng),但毛發(fā)較有光澤,說明益氣排毒方組在改善CRF大鼠一般狀況方面稍優(yōu)于尿毒清組,但無差異。 (2)改善體重方面。正常對照組、尿毒清組和益氣排毒方組體重均逐漸上升,正常組上升幅度較大,尿毒清組上升較慢,空白組大鼠在實驗第二周體重達到峰值,隨即開始下降進一步加重。實驗后,益氣排毒方組與正常對照組、模型對照組和尿毒清組相比差異均具有統(tǒng)計學(xué)意義(P0.05)；尿毒清組與模型對照組相比體重?zé)o差異(P0.05),與益氣排毒方組相比有差異(P0.05),說明尿毒清在改善CRF大鼠體重方面效果不及益氣排毒方。 (3)對腎功能的影響：益氣排毒方與模型對照組相比較,血肌酐和尿素氮的降低均有統(tǒng)計學(xué)意義(P0.05),與正常組相比兩指標(biāo)均顯著升高(P0.01),益氣排毒方與尿毒清相比血肌酐顯著降低(P0.05),尿素氮差異無統(tǒng)計學(xué)意義(P0.05)。(4)對血脂的影響：益氣排毒方組在TG和TC上與正常對照組相比,均無差異(P0.05),而與模型對照組顯著升高(P0.01)。尿毒清組較正常對照組顯著降低(P0.05),較益氣毒方組顯著升高(P0.05)。 (5)對血清鈣、磷的影響：益氣排毒方組血清鈣與正常對照組相比無顯著差異(P0.05),與模型對照組比有顯著差異(P0.01),與尿毒清相比顯著升高(P0.05)；調(diào)節(jié)血磷方面,益氣排毒方與正常對照組、模型對照組和尿毒清組相比,差異均無統(tǒng)計學(xué)意義(P0.05)。 (6)對血紅蛋白的影響：益氣排毒方組血紅蛋白與正常組無明顯差異(P0.05),與模型對照組和尿毒清組相比明顯升高(P0.01)。在紅細(xì)胞計數(shù)方面,益氣排毒方組與正常對照組、模型對照組和尿毒清組相比,均有較大差異(P0.01)。 (7)腎臟病理觀察： ①腎臟外觀觀察上,益氣排毒方組腎臟腫脹、薄膜增厚程度較模型對照組輕,表面顏色較模型對照組深,說明該方具有保護腎功能的作用。 ②對腎重/體重比的影響。腺嘌呤至大鼠腎功能衰竭,出現(xiàn)腎臟腫脹,腎重/體重比值明顯升高,與正常組比較具有顯著差異(P0.01),益氣排毒方能減輕腎臟損害水腫的進程,降低腎重/體重比值(P0.01),這種作用優(yōu)于尿毒清組(P0.05)。 ③HE染色觀察：益氣排毒方組大鼠腎臟有少量腎小管上皮細(xì)胞脫落,部分腎小管擴張,部分可見深黃色結(jié)晶,其腎臟損害的表現(xiàn)較空白組輕。 結(jié)論： 益氣排毒方對腺嘌呤所致慢性腎功能不全大鼠具有改善大鼠體重降低,改善一般情況,改善腎功能,調(diào)節(jié)血脂,糾正貧血,糾正鈣磷代謝紊亂的作用。在降低血肌酐、血脂和調(diào)節(jié)鈣磷代謝方面,與尿毒清相比療效更好,具有良好的發(fā)展前景,但其具體作用機制還有待進一步深入探討。
[Abstract]:Chronic renal insufficiency, also known as chronic kidney disease, is a clinical syndrome characterized by metabolite retention, water, electrolyte, acid-base imbalance and systemic involvement. Untimely and inappropriate treatment will lead to accumulation of toxins in the body and increase the damage to kidney function, thus rapidly entering the uremic period. The accumulation of uremic toxins will seriously endanger the life and health of patients and reduce the quality of life. The huge cost of treatment is a huge burden to families and society. Therefore, exploring the treatment of chronic renal failure. The whole new method is very clinically significant.
Yiqi Paidu Recipe is a summary of Professor Ouyang Yonghong's clinical experience in more than 30 years in Qingyuan Hospital of Traditional Chinese Medicine. It is mainly used in the treatment of patients with renal impairment and elevated serum urea nitrogen and creatinine, that is, patients with chronic renal insufficiency. In 1992, Professor Ouyang Yonghong proposed to use the method of "supplementing Qi and reducing turbidity" to treat chronic renal insufficiency. In 2011, Wang Rui and other studies found that Yiqi Paidu Recipe can intervene in maintaining the Micro-inflammatory State of hemodialysis patients and reduce micro-inflammation. In order to clarify the mechanism of Yiqi Paidu Recipe in improving chronic renal insufficiency, we demonstrated that Yiqi Paidu Recipe is improving the chronic renal insufficiency through animal experiments. In general, it can protect the renal function of rats, regulate the metabolism of blood lipids, calcium and phosphorus, improve anemia and so on.
Objective:
Adenine was used to poison the kidney of rats to establish the model of chronic renal insufficiency. The effects of Yiqi Paidu Recipe on the general condition of rats with chronic renal insufficiency, the indexes of serum creatinine, urea nitrogen, hemoglobin, blood lipids, blood calcium and phosphorus metabolism were observed, and the pathological changes of kidney were observed. To analyze the mechanism of action and explore the mechanism of Yiqi detoxification formula in correcting chronic renal failure.
Method:
Forty Wistar rats of 180-220 g SPF grade were divided into four groups randomly after one week of regular feeding. One group was normal control group, the other 30 were chronic renal insufficiency model group. Adenine was given 100mg/kg/d by gastric lavage. After 21 days of successful modeling, the rats were fed with solid diet and free drinking water. Rats were randomly divided into three groups: model control group, Niao Du Qing group, Yiqi Paidu Fang group. The rats were converted according to the weight of normal adults and rats. The rats in Niu Du Qing group were given intragastric administration according to the ratio of 4.0g/kg, and the rats in Yiqi Paidu Fang group were given intragastric administration according to 23.38ml/kg. To the stomach, an average of about 4 ml of liquid per rat, continuous administration for 28 days. Anesthesia, blood samples, serum creatinine, urea nitrogen, blood lipids, electrolytes and other indicators to observe the pathological changes of the two kidneys.
Result:
1. Modeling results: Compared with the normal group, the rats in the model group had worse mental state, sluggish reaction, grey-white hair and less activity. There was a significant difference in body weight (P 0.05), but there was no significant difference in body weight between the model group (P 0.05). Creatinine and urea nitrogen increased significantly, renal pathological anatomy showed enlarged renal volume, pale color, showing a "large white kidney", HE staining showed glomerulus, renal tubules, renal interstitial tissue damage.
2, treatment outcome:
(1) General condition. Rats with chronic renal insufficiency who were given Yiqi Paidu Recipe were generally better than those in the model control group. Rats in Yiqi Paidu Recipe group had better mental state, quick reaction, more food intake, glossy hair, but not yet reached the normal level. The luster showed that Yiqi Paidu Fang group was slightly better than Niaoduqing group in improving the general condition of CRF rats, but there was no difference.
(2) Improvement of body weight. In the normal control group, the body weight of Niaoduqing group and Yiqi Paidu Fang group increased gradually, while that of the normal group increased greatly, and that of the Niaoduqing group increased slowly. In the blank group, the body weight of the rats reached the peak value in the second week of the experiment, and then began to decline further. After the experiment, the Yiqi Paidu Fang group and the normal control group, the model control group and the model control group. Compared with the model control group, there was no difference in body weight between the two groups (P 0.05), and there was a difference between the two groups (P 0.05), indicating that the effect of Niaoduqing on improving the body weight of CRF rats was not as good as that of Yiqi Paidu Recipe.
(3) Effect on kidney function: Compared with model control group, Yiqi Paidu Recipe had statistical significance in reducing serum creatinine and urea nitrogen (P 0.05), both indexes were significantly higher than normal group (P 0.01), Yiqi Paidu Recipe had significantly lower serum creatinine (P 0.05) and no significant difference in urea nitrogen (P 0.05). Effect: Compared with the normal control group, the TG and TC of the Yiqi Paidu Formula group had no difference (P 0.05), but increased significantly (P 0.01) compared with the model control group.
(5) Effect of Yiqi Paidu Recipe on serum calcium and phosphorus: There was no significant difference between Yiqi Paidu Recipe group and normal control group (P Academic meaning (P0.05).
(6) Effect of Yiqi Paidu Recipe on hemoglobin: There was no significant difference between Yiqi Paidu Recipe group and normal group (P 0.05), but it was significantly higher than model control group and Niaoduqing group (P 0.01). In red blood cell count, Yiqi Paidu Recipe group was significantly different from normal control group, model control group and Niaoduqing group (P 0.01).
(7) renal pathology:
(1) Kidney appearance observation showed that Yiqi Paidu Recipe could protect the kidney function because of the swelling of the kidney, the thickness of the membrane was lighter than that of the model control group, and the surface color was darker than that of the model control group.
(2) Effect on the ratio of kidney weight to body weight.Adenine-induced renal failure in rats resulted in kidney swelling and the ratio of kidney weight to body weight increased significantly compared with the normal group (P 0.01). Yiqi Paidu Recipe could reduce the process of kidney edema and the ratio of kidney weight to body weight (P 0.01), which was better than that of Uremic Qing (P 0.05).
(3) HE staining observation: the kidney of rats in Yiqi Paidu Recipe group showed a small amount of renal tubular epithelial cell exfoliation, some renal tubules dilatation, some dark yellow crystals, and the renal damage was lighter than the blank group.
Conclusion:
Yiqi Paidu Recipe can improve the weight loss of rats with chronic renal insufficiency induced by adenine, improve general conditions, improve renal function, regulate blood lipids, correct anemia, correct calcium and phosphorus metabolism disorders. However, its specific mechanism remains to be further explored.
【學(xué)位授予單位】：廣州中醫(yī)藥大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R277.5
本文編號：2211991
[Abstract]:Chronic renal insufficiency, also known as chronic kidney disease, is a clinical syndrome characterized by metabolite retention, water, electrolyte, acid-base imbalance and systemic involvement. Untimely and inappropriate treatment will lead to accumulation of toxins in the body and increase the damage to kidney function, thus rapidly entering the uremic period. The accumulation of uremic toxins will seriously endanger the life and health of patients and reduce the quality of life. The huge cost of treatment is a huge burden to families and society. Therefore, exploring the treatment of chronic renal failure. The whole new method is very clinically significant.
Yiqi Paidu Recipe is a summary of Professor Ouyang Yonghong's clinical experience in more than 30 years in Qingyuan Hospital of Traditional Chinese Medicine. It is mainly used in the treatment of patients with renal impairment and elevated serum urea nitrogen and creatinine, that is, patients with chronic renal insufficiency. In 1992, Professor Ouyang Yonghong proposed to use the method of "supplementing Qi and reducing turbidity" to treat chronic renal insufficiency. In 2011, Wang Rui and other studies found that Yiqi Paidu Recipe can intervene in maintaining the Micro-inflammatory State of hemodialysis patients and reduce micro-inflammation. In order to clarify the mechanism of Yiqi Paidu Recipe in improving chronic renal insufficiency, we demonstrated that Yiqi Paidu Recipe is improving the chronic renal insufficiency through animal experiments. In general, it can protect the renal function of rats, regulate the metabolism of blood lipids, calcium and phosphorus, improve anemia and so on.
Objective:
Adenine was used to poison the kidney of rats to establish the model of chronic renal insufficiency. The effects of Yiqi Paidu Recipe on the general condition of rats with chronic renal insufficiency, the indexes of serum creatinine, urea nitrogen, hemoglobin, blood lipids, blood calcium and phosphorus metabolism were observed, and the pathological changes of kidney were observed. To analyze the mechanism of action and explore the mechanism of Yiqi detoxification formula in correcting chronic renal failure.
Method:
Forty Wistar rats of 180-220 g SPF grade were divided into four groups randomly after one week of regular feeding. One group was normal control group, the other 30 were chronic renal insufficiency model group. Adenine was given 100mg/kg/d by gastric lavage. After 21 days of successful modeling, the rats were fed with solid diet and free drinking water. Rats were randomly divided into three groups: model control group, Niao Du Qing group, Yiqi Paidu Fang group. The rats were converted according to the weight of normal adults and rats. The rats in Niu Du Qing group were given intragastric administration according to the ratio of 4.0g/kg, and the rats in Yiqi Paidu Fang group were given intragastric administration according to 23.38ml/kg. To the stomach, an average of about 4 ml of liquid per rat, continuous administration for 28 days. Anesthesia, blood samples, serum creatinine, urea nitrogen, blood lipids, electrolytes and other indicators to observe the pathological changes of the two kidneys.
Result:
1. Modeling results: Compared with the normal group, the rats in the model group had worse mental state, sluggish reaction, grey-white hair and less activity. There was a significant difference in body weight (P 0.05), but there was no significant difference in body weight between the model group (P 0.05). Creatinine and urea nitrogen increased significantly, renal pathological anatomy showed enlarged renal volume, pale color, showing a "large white kidney", HE staining showed glomerulus, renal tubules, renal interstitial tissue damage.
2, treatment outcome:
(1) General condition. Rats with chronic renal insufficiency who were given Yiqi Paidu Recipe were generally better than those in the model control group. Rats in Yiqi Paidu Recipe group had better mental state, quick reaction, more food intake, glossy hair, but not yet reached the normal level. The luster showed that Yiqi Paidu Fang group was slightly better than Niaoduqing group in improving the general condition of CRF rats, but there was no difference.
(2) Improvement of body weight. In the normal control group, the body weight of Niaoduqing group and Yiqi Paidu Fang group increased gradually, while that of the normal group increased greatly, and that of the Niaoduqing group increased slowly. In the blank group, the body weight of the rats reached the peak value in the second week of the experiment, and then began to decline further. After the experiment, the Yiqi Paidu Fang group and the normal control group, the model control group and the model control group. Compared with the model control group, there was no difference in body weight between the two groups (P 0.05), and there was a difference between the two groups (P 0.05), indicating that the effect of Niaoduqing on improving the body weight of CRF rats was not as good as that of Yiqi Paidu Recipe.
(3) Effect on kidney function: Compared with model control group, Yiqi Paidu Recipe had statistical significance in reducing serum creatinine and urea nitrogen (P 0.05), both indexes were significantly higher than normal group (P 0.01), Yiqi Paidu Recipe had significantly lower serum creatinine (P 0.05) and no significant difference in urea nitrogen (P 0.05). Effect: Compared with the normal control group, the TG and TC of the Yiqi Paidu Formula group had no difference (P 0.05), but increased significantly (P 0.01) compared with the model control group.
(5) Effect of Yiqi Paidu Recipe on serum calcium and phosphorus: There was no significant difference between Yiqi Paidu Recipe group and normal control group (P Academic meaning (P0.05).
(6) Effect of Yiqi Paidu Recipe on hemoglobin: There was no significant difference between Yiqi Paidu Recipe group and normal group (P 0.05), but it was significantly higher than model control group and Niaoduqing group (P 0.01). In red blood cell count, Yiqi Paidu Recipe group was significantly different from normal control group, model control group and Niaoduqing group (P 0.01).
(7) renal pathology:
(1) Kidney appearance observation showed that Yiqi Paidu Recipe could protect the kidney function because of the swelling of the kidney, the thickness of the membrane was lighter than that of the model control group, and the surface color was darker than that of the model control group.
(2) Effect on the ratio of kidney weight to body weight.Adenine-induced renal failure in rats resulted in kidney swelling and the ratio of kidney weight to body weight increased significantly compared with the normal group (P 0.01). Yiqi Paidu Recipe could reduce the process of kidney edema and the ratio of kidney weight to body weight (P 0.01), which was better than that of Uremic Qing (P 0.05).
(3) HE staining observation: the kidney of rats in Yiqi Paidu Recipe group showed a small amount of renal tubular epithelial cell exfoliation, some renal tubules dilatation, some dark yellow crystals, and the renal damage was lighter than the blank group.
Conclusion:
Yiqi Paidu Recipe can improve the weight loss of rats with chronic renal insufficiency induced by adenine, improve general conditions, improve renal function, regulate blood lipids, correct anemia, correct calcium and phosphorus metabolism disorders. However, its specific mechanism remains to be further explored.
【學(xué)位授予單位】：廣州中醫(yī)藥大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R277.5
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