温补肾阳法对肾阳虚肺纤维化大鼠肺内CTGF表达的影响

The expression of CTGF in the lungs of rats with kidney yang deficiency of pulmonary fibrosis by warming and Tonifying Kidney Yang method

目的探讨温补肾阳法治疗肾阳虚肺纤维化(PF)的作用及机制。方法 60只雄性6周龄Wistar大鼠采用随机数字表法分为正常组、模型组和干预组,采用一次性博来霉素A5气管插管给药加腺嘌呤连续灌胃28 d复制肾阳虚PF大鼠模型。正常组大鼠气管插管给予生理盐水加等渗生理盐水灌胃作为对照,模型组大鼠造模后生理盐水灌胃14 d,干预组大鼠造模后加味参附汤灌胃14 d,每组于实验第42天处死大鼠,留24 h尿液检测17-羟皮质类固醇(17-OH-CS)含量,同时,HE和MASSON染色观察肺纤维化程度,免疫组化法和Western印迹测定大鼠肺内结缔组织生长因子(CTGF)蛋白的表达,RT-PCR检测肺组织CTGF mRNA的表达。结果与正常组比较,肾阳虚肺纤维化模型大鼠尿17-OH-CS明显下降,模型大鼠肺组织内可见肺泡结构大量破坏,大量胶原沉积,干预组大鼠肺泡结构也基本消失,可见较多的胶原沉积,病理组织学表现明显轻于模型组。与正常组比较,模型组肺组织CTGF在肺内的表达水平明显升高(P<0.05);与模型组比较,干预组大鼠肺内CTGF的表达降低(P<0.05)。结论肾阳虚PF大鼠肾阳虚症状明显,肺内纤维化表现明显,造模成功,加味参附汤可以改善大鼠肾阳虚症状,可能通过抑制CTGF蛋白表达而缓解肺内胶原的进一步沉积减缓肺纤维化的发展。

Objective To observe the expression of CTGF in the lungs of rats with kidney yang deficiency of pulmonary fibrosis by warming and Tonifying Kidney Yang method. Methods 60 6-week-old male Wistar rats were randomly divided into normal,model and intervention groups. The rats of normal and model groups were given saline irrigation stomach,the rats of intervention group were given modified Shenfu decoction for 14 d. The content of 17-hydroxyl cortex steroids( 17-OHCS) of 24 h urine was measured,lung fiber was observed by HE and MASSON staining,the expressions of CTGF protein and mRNA in lung tissue were tested by Western blot and RT-PCR. Results Compared with that of control group,and the content of 17-OHCS in model group was lower,decreased alveolar structures of kidney-Yang deficiency model of pulmonary fibrosis rats were destroyed,a large number of collagen was deposited; alveolar structures in intervention group were also disappeared,more collagen was deposited,but the pathological histology changes were significantly lighter than those of model group. Compared with that of control group,the expression of CTGF in pulmonary tissue of model group was increased( P < 0. 05),and it was lower in intervention group than that in model group( P < 0. 05). Conclusions Modified Shenfu decoction could improve the symptoms of kidney yang deficiency rats,and inhibit the expression of CTGF,relief further deposition of collagen in lung,slow down the development of pulmonary fibrosis.