Yhe Effect Of Gouqi And Danshen Compound Extract On RHO、XBP1 And Caspase12 In The RCS Rat With Syndromes Of Blood Stasis And Deficiency

Objective:1.To study the etiology and pathology of TCM about retinitis pigmentosa.2. To observe the changes on syndromes of blood stasis and deficiency in the RCS rat and observe the changes on retinal tissue of tonifying the kidney and promoting blood circulation for treatment of deficiency and blood stasis type retinitis pigmentosa. 3. To study the effect of Gouqi(Lycium barbarum) and Danshen(Salvia miltiorrhiza) compound extract on RHO 、 XBP1 and Caspase12 in the RCS rat with syndromes of blood stasis and deficiency and to explore its mechanism, to provide more evidence for the modern medicine in the treatment of retinitis pigmentosa, to explore the curative effect of the treatment of retinitis pigmentosa and mechanism of tonifying the kidney and promoting blood circulation, to explore the curative effect of the treatment of retinitis pigmentosa and mechanism of tonifying the kidney and promoting blood circulation.Methods:Part 1:20 RCS rats were divided into normal group and model group(10 each). Normal group: female RCS(rdy+/+,p+/+)rats(n=5), male RCS(rdy+/+,p+/+)rats(n=5).Model group: female RCS(rdy-/-,p-/-)rats(n=5), male RCS(rdy-/-,p-/-)rats(n=5). After the model is successfully established, detect the expression of α-SMA in the lung tissue by immunohistochemical method, determinate the level of c AMP, c GMP, c AMP/c GMP, PG, E2, P-Selectin by ELISA, observe the change of hemorheology, and carry on pathologicalexamination about retinal tissue.Part 2:40 RCS rats were divided into normal group, model group, Gouqi group, Danshen group, Gouqi and Danshen group(8 each). Normal group: female RCS(rdy+/+,p+/+)rats(n=4), male RCS(rdy+/+,p+/+)rats(n=4),filling the stomach with 0.9%NS. Model group: female RCS(rdy-/-,p-/-)rats(n=4), male RCS(rdy-/-,p-/-)rats(n=4), filling the stomach with 0.9%NS. Gouqi group: female RCS(rdy-/-,p-/-) rats(n=4), male RCS(rdy-/-,p-/-)rats(n=4), filling the stomach with Gouqi((Lycium barbarum). Danshen group: female RCS(rdy-/-,p-/-)rats(n=4), male RCS(rdy-/-,p-/-)rats(n=4), filling the stomach with Danshen((Lycium barbarum). Gouqi and Danshen group: female RCS(rdy-/-,p-/-)rats(n=4), male RCS(rdy-/-,p-/-)rats(n=4), filling the stomach with Gouqi((Lycium barbarum) and Danshen((Lycium barbarum). After filling the stomach 28 days, determinate the level of c AMP、c GMP、c AMP/c GMP、PG、E2、P-Selectin by ELISA, observe the change of hemorheology, and carry on pathological examination by HE stain about retinal tissue.Part 3:40 RCS rats were divided into normal group, model group, Gouqi group, Danshen group, Gouqi and Danshen group(8 each). Normal group: female RCS(rdy+/+,p+/+)rats(n=4), male RCS(rdy+/+,p+/+)rats(n=4),filling the stomach with 0.9%NS. Model group: female RCS(rdy-/-,p-/-)rats(n=4), male RCS(rdy-/-,p-/-)rats(n=4), filling the stomach with 0.9%NS.Gouqi group: female RCS(rdy-/-,p-/-) rats(n=4), male RCS(rdy-/-,p-/-)rats(n=4), filling the stomach with Gouqi((Lycium barbarum). Danshen group: female RCS(rdy-/-,p-/-)rats(n=4), male RCS(rdy-/-,p-/-)rats(n=4), filling the stomach with Danshen((Lycium barbarum). Gouqi and Danshen group: female RCS(rdy-/-,p-/-)rats(n=4), male RCS(rdy-/-,p-/-)rats(n=4), filling the stomach with Gouqi((Lycium barbarum) and Danshen((Lycium barbarum). After filling the stomach 28 days, detect the expression of RHO, XBP1, and Caspase 12 in the retinal tissue by immunohistochemical method, detect retinal tissue RHO, XBP1, and Caspase 12 proteins relative content by Western blot, determinate the expression of RHO m RNA, XBP1 m RNA, and Caspase 12 m RNA by Real-time q PCR.Results:Part 1:In the female RCS(rdy-/-,p-/-)rats, the content of E2 compared with the normal group were significantly reduced(P<0.05). In the male RCS(rdy-/-,p-/-)rats, the content of PG compared with the normal group were significantly reduced(P<0.01). Compared with the normal group, the contents of c AMP、c GMP、c AMP/c GMP in the retinal homogenate of the RCS(rdy-/-,p-/-)rats were significantly increased(P<0.01). Compared with the normal group, the level of blood viscosity(5.00/s, 100.00/s, 200.00/s), plasma viscosity, high shear rate, low shear rate, and plasma fibrinogen in the blood of the RCS(rdy-/-,p-/-)rats were significantly increased(P<0.05). Compared with the model group, the level of P-selectin in the blood of the RCS(rdy-/-,p-/-) rats were significantly increased(P<0.01). The slice of model group retinal pathology organization dyed by HE showed that there are disorder of retinal outer structure.Part 2:In the female Gouqi and Danshen group, the content of E2 compared with the model group were significantly increased(P <0.01). In the male Gouqi and Danshen group, the content of PG compared with the normal group were significantly increased(P <0.01). Compared with the model group, the contents of c AMP、c GMP、c AMP/c GMP in the retinal homogenate of the RCS(rdy-/-,p-/-) rats in Gouqi and Danshen group were significantly reduced(P<0.05). Compared with the model group, the level of plasma viscosity, high shear rate, and plasma fibrinogen in the blood of the RCS(rdy-/-,p-/-) rats in Gouqi and Danshen group were significantly reduced(P<0.05). Compared with the model group, the level of P-selectin in the blood of the RCS(rdy-/-,p-/-) rats in Gouqi and Danshen group were significantly reduced(P <0.01). The slice of retinal pathology organization dyed by HE of the RCS(rdy-/-,p-/-) rats in Gouqi and Danshen group showed that there are improvement on disorder of retinal outer structure.Part 3:Compared with the normal group, model group rats XBP1 and Caspase 12 m RNA expression increased significantly(P<0.01). Compared with model group, Gouqi and Danshen group can significantly reduce the expression levels of XBP1 and Caspase 12 m RNA(P < 0.01). Compared with the normal group, model group rats RHO m RNA expression reduced significantly(P < 0.01). Compared with model group, Gouqi and Danshen group can significantly increase the expression levels of RHO m RNA(P<0.05). Compared with the normal group, model group rats XBP1 protein and Caspase 12 protein expression also increased significantly(P<0.01). Compared with model group, Gouqi and Danshen group can significantly reduce the expression levels of XBP1 protein and Caspase 12 protein(P<0.05). Compared with the normal group, model group rats RHO protein expression reduced significantly(P < 0.01). Compared with model group, Gouqi and Danshen group can significantly increase the expression levels of RHO protein(P<0.01). The expression of XBP1 positively correlated with the expression of Caspase 12(r=0.512,P=0.001, P<0.01). The expression of RHO negatively correlated with the expression of Caspase 12(r=-0.588,P=0.000, P<0.01). The expression of RHO negatively correlated with the expression of XBP1(r=-0.597,P=0.000, P<0.01).Conclusion:The following conclusions were drawn through the experimental research: The etiology and pathology of TCM about retinitis pigmentosa in the RCS(rdy-/-,p-/-)rats is congenital deficiency with blood stasis.The RCS(rdy-/-,p-/-)rats, systemic symptoms of blood stasis and deficiency can be improved with Gouqi and Danshen filled the rats, stomach. Gouqi and Danshen can significantly reduce endoplasmic reticulum stress and reduce the cell apoptosis in the retinal tissue,and enhance the expression of rhodopsin. So that the principle of RP with syndromes of blood stasis and deficiency is activating the blood circulation to dissipate blood stasis and invigorate the deficiency.