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黄连素对糖尿病大鼠牙槽骨吸收的影响

Effect of berberine on alveolar bone absorption in diabetic rats

摘要:

目的:研究黄连素干预对糖尿病大鼠牙槽骨吸收的影响及可能机制。方法:通过一次性腹腔注射链脲佐菌素(35 mg/kg)建立糖尿病大鼠模型。将60只健康无特定病原体级雌性Sprague-Dawley大鼠按随机数字表法分为正常组、糖尿病组和黄连素组,每组20只。黄连素组和糖尿病组大鼠建立糖尿病模型后分别给予黄连素(200 mg/kg)和等体积的氯化钠注射液灌胃,每天1次,连续灌胃4周;正常组大鼠不予任何处理。检测大鼠的空腹血糖、空腹胰岛素(Fins)水平,计算胰岛素抵抗指数(HOMA-IR)和胰岛素敏感指数(ISI),并检测大鼠的血清钙、磷、碱性磷酸酶(ALP)和骨钙素水平。苏木精-伊红染色观察大鼠牙周组织的病理学变化,反转录PCR和蛋白质印迹法(Western Blot)检测大鼠牙槽骨组织中骨形态发生蛋白-2(BMP-2)、Smad1、Smad4基因的mRNA和蛋白表达变化。结果:与糖尿病组相比,黄连素组大鼠空腹血糖水平[(9.88±1.23)mmol/L比(21.66±2.41)mmol/L]、Fins水平[(11.63±3.63)mIU/L比(21.64±5.58)mIU/L]和HOMA-IR(7.58±0.96比20.85±1.95)均降低,ISI(-5.10±0.16比-6.15±0.18)以及血清钙[(1.855±0.125)mmol/L比(1.317±0.104)mmol/L]、磷[(3.087±0.278)mmol/L比(2.526±0.226)mmol/L]、ALP[(130.681±18.963)U/L比(99.874±31.968)U/L]和骨钙素水平[(112.368±9.752)ng/ml比(78.645±10.973)ng/ml]均升高,差异均具有统计学意义(均 P<0.05)。苏木精-伊红染色结果显示,正常组大鼠牙槽骨未见骨质吸收,糖尿病组大鼠牙槽骨可见骨质吸收,而黄连素组大鼠牙槽骨骨质吸收较糖尿病组明显减轻。反转录PCR和Western Blot结果显示,与糖尿病组相比,黄连素组大鼠牙槽骨组织中BMP-2、Smad1、Smad4基因的mRNA和蛋白表达均明显升高(均 P<0.05)。 结论:黄连素干预可降低糖尿病大鼠的空腹血糖水平,并改善胰岛素抵抗,其可能通过上调BMP-2/Smad信号通路减轻糖尿病大鼠的牙槽骨吸收。

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abstracts:

Objective:To study the effect and mechanism of berberine intervention on alveolar bone absorption in diabetic rats.Methods:Diabetes rat model was established by one-time intraperitoneal injection of streptozotocin (35 mg/kg). Sixty healthy female Sprague-Dawley rats without specific pathogens were divided into normal group, diabetic group and berberine group according to the random number table method, 20 rats for each group. The rat models in the berberine group and the diabetic group were given berberine (200 mg/kg) and an equal volume of sodium chloride injection by gavage, once a day, for 4 consecutive weeks. The rat models in the normal group were not treated. Fasting blood glucose (FBG), fasting insulin level (Fins) , hormone resistance index (HOMA-IR) and insulin sensitivity index (ISI), as well as the serum calcium, phosphorus content, alkaline phosphatase activity and osteocalcin content of the rats were measured. The pathological changes of periodontal tissues were detected by HE staining. The expression of BMP-2/Smad pathway (BMP-2, Smad1, Smad4) was detected by RT-PCR and Western blot.Results:Compared with the diabetes group, the FBG level of rats in the berberine group [(9.88±1.23) mmol/L vs. (21.66±2.41) mmol/L], Fins levels [(11.63±3.63) mIU/L vs. (21.64±5.58) mIU/L] , HOMA-IR (7.58±0.96 vs. 20.85±1.95) were reduced, and ISI (-5.10±0.16 vs. 6.15±0.18), serum calcium levels[(1.855±0.125) mmol/L vs. (1.317±0.104)) mmol/L], serum phosphorus levels [(3.087±0.278) mmol/L vs. (2.526±0.226) mmol/L], serum ALP levels [(130.681±18.963) U/L vs. (99.874±31.968) U/L] and osteocalcin levels [(112.368±9.752) ng/ml vs. (78.645±10.973) ng/ml] were all increased, and the differences were statistically significant (all P<0.05). The results of hematoxylin-eosin staining showed that there was no bone resorption in the alveolar bone of the rats in the normal group, bone resorption was found in the alveolar bone of the rats in the diabetes group, and the bone resorption of the alveolar bone in the berberine group was significantly reduced. RT-PCR and Western Blot results showed that compared with the diabetes group, the BMP-2, Smad1, Smad4 gene mRNA and protein in the alveolar bone tissue of the berberine group were significantly increased (all P<0.05). Conclusions:Berberine intervention can reduce FBG levels in diabetic rats and improve insulin resistance, which may reduce alveolar bone resorption in diabetic rats by up-regulating the BMP-2/Smad signaling pathway.

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