清胆泻热法治疗鼻渊机理研究
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摘要
目的:通过动物实验在分子生物学、形态学等方面,从基因水平上探讨清胆泻热法治疗鼻渊的作用机制,从而更准确地指导临床用药。
方法:将健康SD大鼠随机分为①空白组;②模型组:③中药低剂量组;④中药中剂量组;⑤中药高剂量组;⑥青霉素Ⅴ钾组。急性鼻-鼻窦炎模型依据叶菁报道的方法造模。造模7天后,空白组和模型组按5ml·kg~(-1)·d~(-1)灌服无菌水,其余各组分别灌服相应的药物。从造模第1天开始,观察动物一般情况变化,pH值、体温及体重变化等,着重观察鼻部症状、鼻分泌量。用药7天后,处死动物,每组随机选取6只大鼠。用ELISA法检测各组大鼠血清中C-反应蛋白(CRP)浓度。在超净工作台上取鼻黏膜后,提取mRNA,并逆转录为cDNA,用RT-PCR法测定白细胞介素2mRNA(IL-2 mRNA)、白细胞介素4 mRNA(IL-4 mRNA)、白细胞介素10 mRNA(IL-10 mRNA)、干扰素-γ(IFN-γmRNA)的表达.在透射电镜下观察鼻黏膜的病理形态变化。
结果:
1.模型组大鼠鼻腔分泌物pH值与空白组相比,有显著性升高(P＜0.01)。用药7天后,各治疗组药物均能降低鼻腔分泌物pH值。其中以中药低剂量组效果最明显(P＜0.01)。
2.模型组大鼠血清CRP浓度与空白组相比,有显著性升高(P＜0.01)。用药7天后,各治疗组药物均能降低血清CRP浓度。其中以中药低、中剂量组效果最明显(P＜0.01).
3.用药7天后,除中药高剂量组外,其余各治疗组药物均能使鼻黏膜组织IL-2 mRNA及IL-10 mRNA的相对表达量升高,以中药低剂量组及青霉素Ⅴ钾组效果最明显(P＜0.01)。
4.用药7天后,除中药高剂量组外,其余各治疗组药物均能使鼻黏膜组织IL-4 mRNA的相对表达量升高,以中药低剂量组效果最明显(P＜0.01)。
5.用药7天后,各治疗组物均能使鼻黏膜组织的IFN-γmRNA相对表达量降低,以中药低、中剂量组效果最明显(P＜0.01)。
6.用药7天后,透射电镜观察显示:除中药高剂量组外,其余各治疗组药物均能使鼻黏膜纤毛形态结构逐步恢复正常,以中药低、中剂量组效果最明显。
结论:
1.清胆泻热可以降低鼻分泌物pH值至接近正常值,以保证保鼻腔内环境的协调。
2.清胆泻热可以降低血清CRP浓度,抑制或杀死病原微生物,消除炎症反应.
3.清胆泻热可以增强鼻黏膜IL-2 mRNA的表达,作用于T细胞,增强细胞免疫,提高机体抗感染能力.
4.清胆泻热可以增强鼻黏膜IL-4 mRNA和IL-10 mRNA的表达,发挥其抗炎活性.
5.清胆泻热可以抑制鼻黏膜IFN-γmRNA的表达,从而抑制相关炎性因子的合成。
6.清胆泻热可以减轻内质网扩张、线粒体肿胀,促进鼻黏液纤毛传输系统恢复正常.
我们推测吉雷开窍汤可能以清胆泻热法通过改善机体内环境:扶助正气、驱邪外出,发挥治疗作用。
Objective:To study the mechanism of action about the method of clearing gallbladder and purging heat to Bi Yuan at the gene level on molecular biology and molecular biology by animal experiment,then to guide the clinical medication more accurately.
Methods:Group the healthy SD rats into the following six parts randomly:blank group,model group,Ji Lei Kai Qiao Tang low-dose group,Ji Lei Kai Qiao Tang middle-dose,Ji Lei Kai Qiao Tang high-dose and penicillinum v potassium group. According to the method reported by YE Jing we made the Acute Rhinosinusitis model.In the following seven days we drenched the blank group and the model group 5ml·kg~(-1)·d~(-1) with the sterile water(SW),and the other four groups with corresponding medicine.All these days we observed the changes of the rats about the pH value,body temperature,and body weight,especially the nose symptom and secretory volume. After seven days we killed these rats and chose six in each group randomly.We measured the C-reactive protein(CRP) concentration in the SD rats' blood serum using the ELISA method.First we took out of the mucous membrane of nose at the superclean bench,after that we extracted the mRNA and reversed transcription to cDNA,then we measured the expressions of IL-2 mRNA,IL-4 mRNA,IL-10 mRNA and IFN-γmRNA by using the RT-PCR method.At last we observed the morphologic changes of the mucous membrane of nose under the electron microscope.
Results:
1.Comparing with the blank group,the pH value of the model group's nasal cavity secretion substance has improved significantly(P＜0.01).The nasal cavity secretion substance's pH values of all the therapy groups have declined,especially the Ji Lei Kai Qiao Tang low-dose group.
2.Comparing with the blank group,the blood serum CRP concentration of the model group has improved significantly(P＜0.01).The blood serum CRP concentrations of all the therapy groups have declined especially the Ji Lei Kai Qiao Tang low-dose group and the middle-dose group(P＜0.01).
3.After seven days medication,except the Ji Lei Kai Qiao Tang high-dose group, the expressions of IL-2 mRNA and IL-10 mRNA of all the therapy groups have improved especially the Ji Lei Kai Qiao Tang low-dose group and the penicillinum v potassium group(P＜0.01).
4.After seven days medication,except the Ji Lei Kai Qiao Tang high-dose group, the expressions of IL-4 mRNA of all the therapy groups have improved especially the Ji Lei Kai Qiao Tang low-dose group(P＜0.01).
5.After seven days medication,the expressions of IFN-γmRNA of all the therapy groups have declined especially the Ji Lei Kai Qiao Tang low-close group and the middle-dose group(P＜0.01).
6.After seven days medication,through the transmission electron microscope we find that except the Ji Lei Kai Qiao Tang high-dose group,the membrane of nose cilia of all the therapy groups have returned to normal gradually especially the Ji Lei Kai Qiao Tang low-dose group and the middle-dose group.
Conclusion:
1.By falling the Nasal discharge pH value to normal,clearing gallbladder and purging heat can make the internal environment concordance of the nasal cavity.
2.Clearing gallbladder and purging heat can reduce the CRP concentration of the blood serum,depression or kill the causative organism,and eliminate the inflammatory reaction.
3.Clearing gallbladder and purging heat can strengthen the expression of the nasal mucosa IL-2 mRNA,act on the T cell to strengthen the cell immunity and improve the anti-infection ability of the organism.
4.Clearing gallbladder and purging heat can strengthen the expression of the nasal mucosa IL-4 mRNA and IL-10 mRNA,as a result to educe the anti-inflammatory activity.
5.Clearing gallbladder and purging heat can restrain the expression of the nasal mucosa IFN-γmRNA,as a result to restrain the synthesization of the correlated inflammatory factor.
6.Clearing gallbladder and purging heat can lighten the broaden of the endocytoplasmic reticulum and the engorgement of the mitochondria,as a result to promote the nasal Mucus cilia transmission system to return to normal.
We suppose that Ji Lei Kai Qiao Tang can improve organism internal environment by the method of clearing gallbladder and purging heat,strengthening vital qi to eliminate pathogenic factor.
方法:将健康SD大鼠随机分为①空白组;②模型组:③中药低剂量组;④中药中剂量组;⑤中药高剂量组;⑥青霉素Ⅴ钾组。急性鼻-鼻窦炎模型依据叶菁报道的方法造模。造模7天后,空白组和模型组按5ml·kg~(-1)·d~(-1)灌服无菌水,其余各组分别灌服相应的药物。从造模第1天开始,观察动物一般情况变化,pH值、体温及体重变化等,着重观察鼻部症状、鼻分泌量。用药7天后,处死动物,每组随机选取6只大鼠。用ELISA法检测各组大鼠血清中C-反应蛋白(CRP)浓度。在超净工作台上取鼻黏膜后,提取mRNA,并逆转录为cDNA,用RT-PCR法测定白细胞介素2mRNA(IL-2 mRNA)、白细胞介素4 mRNA(IL-4 mRNA)、白细胞介素10 mRNA(IL-10 mRNA)、干扰素-γ(IFN-γmRNA)的表达.在透射电镜下观察鼻黏膜的病理形态变化。
结果:
1.模型组大鼠鼻腔分泌物pH值与空白组相比,有显著性升高(P＜0.01)。用药7天后,各治疗组药物均能降低鼻腔分泌物pH值。其中以中药低剂量组效果最明显(P＜0.01)。
2.模型组大鼠血清CRP浓度与空白组相比,有显著性升高(P＜0.01)。用药7天后,各治疗组药物均能降低血清CRP浓度。其中以中药低、中剂量组效果最明显(P＜0.01).
3.用药7天后,除中药高剂量组外,其余各治疗组药物均能使鼻黏膜组织IL-2 mRNA及IL-10 mRNA的相对表达量升高,以中药低剂量组及青霉素Ⅴ钾组效果最明显(P＜0.01)。
4.用药7天后,除中药高剂量组外,其余各治疗组药物均能使鼻黏膜组织IL-4 mRNA的相对表达量升高,以中药低剂量组效果最明显(P＜0.01)。
5.用药7天后,各治疗组物均能使鼻黏膜组织的IFN-γmRNA相对表达量降低,以中药低、中剂量组效果最明显(P＜0.01)。
6.用药7天后,透射电镜观察显示:除中药高剂量组外,其余各治疗组药物均能使鼻黏膜纤毛形态结构逐步恢复正常,以中药低、中剂量组效果最明显。
结论:
1.清胆泻热可以降低鼻分泌物pH值至接近正常值,以保证保鼻腔内环境的协调。
2.清胆泻热可以降低血清CRP浓度,抑制或杀死病原微生物,消除炎症反应.
3.清胆泻热可以增强鼻黏膜IL-2 mRNA的表达,作用于T细胞,增强细胞免疫,提高机体抗感染能力.
4.清胆泻热可以增强鼻黏膜IL-4 mRNA和IL-10 mRNA的表达,发挥其抗炎活性.
5.清胆泻热可以抑制鼻黏膜IFN-γmRNA的表达,从而抑制相关炎性因子的合成。
6.清胆泻热可以减轻内质网扩张、线粒体肿胀,促进鼻黏液纤毛传输系统恢复正常.
我们推测吉雷开窍汤可能以清胆泻热法通过改善机体内环境:扶助正气、驱邪外出,发挥治疗作用。
Objective:To study the mechanism of action about the method of clearing gallbladder and purging heat to Bi Yuan at the gene level on molecular biology and molecular biology by animal experiment,then to guide the clinical medication more accurately.
Methods:Group the healthy SD rats into the following six parts randomly:blank group,model group,Ji Lei Kai Qiao Tang low-dose group,Ji Lei Kai Qiao Tang middle-dose,Ji Lei Kai Qiao Tang high-dose and penicillinum v potassium group. According to the method reported by YE Jing we made the Acute Rhinosinusitis model.In the following seven days we drenched the blank group and the model group 5ml·kg~(-1)·d~(-1) with the sterile water(SW),and the other four groups with corresponding medicine.All these days we observed the changes of the rats about the pH value,body temperature,and body weight,especially the nose symptom and secretory volume. After seven days we killed these rats and chose six in each group randomly.We measured the C-reactive protein(CRP) concentration in the SD rats' blood serum using the ELISA method.First we took out of the mucous membrane of nose at the superclean bench,after that we extracted the mRNA and reversed transcription to cDNA,then we measured the expressions of IL-2 mRNA,IL-4 mRNA,IL-10 mRNA and IFN-γmRNA by using the RT-PCR method.At last we observed the morphologic changes of the mucous membrane of nose under the electron microscope.
Results:
1.Comparing with the blank group,the pH value of the model group's nasal cavity secretion substance has improved significantly(P＜0.01).The nasal cavity secretion substance's pH values of all the therapy groups have declined,especially the Ji Lei Kai Qiao Tang low-dose group.
2.Comparing with the blank group,the blood serum CRP concentration of the model group has improved significantly(P＜0.01).The blood serum CRP concentrations of all the therapy groups have declined especially the Ji Lei Kai Qiao Tang low-dose group and the middle-dose group(P＜0.01).
3.After seven days medication,except the Ji Lei Kai Qiao Tang high-dose group, the expressions of IL-2 mRNA and IL-10 mRNA of all the therapy groups have improved especially the Ji Lei Kai Qiao Tang low-dose group and the penicillinum v potassium group(P＜0.01).
4.After seven days medication,except the Ji Lei Kai Qiao Tang high-dose group, the expressions of IL-4 mRNA of all the therapy groups have improved especially the Ji Lei Kai Qiao Tang low-dose group(P＜0.01).
5.After seven days medication,the expressions of IFN-γmRNA of all the therapy groups have declined especially the Ji Lei Kai Qiao Tang low-close group and the middle-dose group(P＜0.01).
6.After seven days medication,through the transmission electron microscope we find that except the Ji Lei Kai Qiao Tang high-dose group,the membrane of nose cilia of all the therapy groups have returned to normal gradually especially the Ji Lei Kai Qiao Tang low-dose group and the middle-dose group.
Conclusion:
1.By falling the Nasal discharge pH value to normal,clearing gallbladder and purging heat can make the internal environment concordance of the nasal cavity.
2.Clearing gallbladder and purging heat can reduce the CRP concentration of the blood serum,depression or kill the causative organism,and eliminate the inflammatory reaction.
3.Clearing gallbladder and purging heat can strengthen the expression of the nasal mucosa IL-2 mRNA,act on the T cell to strengthen the cell immunity and improve the anti-infection ability of the organism.
4.Clearing gallbladder and purging heat can strengthen the expression of the nasal mucosa IL-4 mRNA and IL-10 mRNA,as a result to educe the anti-inflammatory activity.
5.Clearing gallbladder and purging heat can restrain the expression of the nasal mucosa IFN-γmRNA,as a result to restrain the synthesization of the correlated inflammatory factor.
6.Clearing gallbladder and purging heat can lighten the broaden of the endocytoplasmic reticulum and the engorgement of the mitochondria,as a result to promote the nasal Mucus cilia transmission system to return to normal.
We suppose that Ji Lei Kai Qiao Tang can improve organism internal environment by the method of clearing gallbladder and purging heat,strengthening vital qi to eliminate pathogenic factor.
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